Chromaffin Granules: Organelles in CHROMAFFIN CELLS located in the adrenal glands and various other organs. These granules are the site of the synthesis, storage, metabolism, and secretion of EPINEPHRINE and NOREPINEPHRINE.Chromaffin System: The cells of the body which stain with chromium salts. They occur along the sympathetic nerves, in the adrenal gland, and in various other organs.Adrenal Medulla: The inner portion of the adrenal gland. Derived from ECTODERM, adrenal medulla consists mainly of CHROMAFFIN CELLS that produces and stores a number of NEUROTRANSMITTERS, mainly adrenaline (EPINEPHRINE) and NOREPINEPHRINE. The activity of the adrenal medulla is regulated by the SYMPATHETIC NERVOUS SYSTEM.Chromaffin Cells: Cells that store epinephrine secretory vesicles. During times of stress, the nervous system signals the vesicles to secrete their hormonal content. Their name derives from their ability to stain a brownish color with chromic salts. Characteristically, they are located in the adrenal medulla and paraganglia (PARAGANGLIA, CHROMAFFIN) of the sympathetic nervous system.Cytoplasmic Granules: Condensed areas of cellular material that may be bounded by a membrane.Cattle: Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.Dopamine beta-HydroxylaseChromogranins: A group of acidic proteins that are major components of SECRETORY GRANULES in the endocrine and neuroendocrine cells. They play important roles in the aggregation, packaging, sorting, and processing of secretory protein prior to secretion. They are cleaved to release biologically active peptides. There are various types of granins, usually classified by their sources.Catecholamines: A general class of ortho-dihydroxyphenylalkylamines derived from tyrosine.Chromogranin A: A type of chromogranin which was first isolated from CHROMAFFIN CELLS of the ADRENAL MEDULLA but is also found in other tissues and in many species including human, bovine, rat, mouse, and others. It is an acidic protein with 431 to 445 amino acid residues. It contains fragments that inhibit vasoconstriction or release of hormones and neurotransmitter, while other fragments exert antimicrobial actions.Exocytosis: Cellular release of material within membrane-limited vesicles by fusion of the vesicles with the CELL MEMBRANE.Adrenal Glands: A pair of glands located at the cranial pole of each of the two KIDNEYS. Each adrenal gland is composed of two distinct endocrine tissues with separate embryonic origins, the ADRENAL CORTEX producing STEROIDS and the ADRENAL MEDULLA producing NEUROTRANSMITTERS.Intracellular Membranes: Thin structures that encapsulate subcellular structures or ORGANELLES in EUKARYOTIC CELLS. They include a variety of membranes associated with the CELL NUCLEUS; the MITOCHONDRIA; the GOLGI APPARATUS; the ENDOPLASMIC RETICULUM; LYSOSOMES; PLASTIDS; and VACUOLES.Chromogranin B: A type of chromogranin which was initially characterized in a rat PHEOCHROMOCYTOMA CELL LINE. It is found in many species including human, rat, mouse, and others. It is an acidic protein with 626 to 657 amino acid residues. In some species, it inhibits secretion of PARATHYROID HORMONE or INSULIN and exerts bacteriolytic effects in others.Annexin A7: An annexin family member that plays a role in MEMBRANE FUSION and signaling via VOLTAGE-DEPENDENT CALCIUM CHANNELS.Tetrabenazine: A drug formerly used as an antipsychotic and treatment of various movement disorders. Tetrabenazine blocks neurotransmitter uptake into adrenergic storage vesicles and has been used as a high affinity label for the vesicle transport system.Catecholamine Plasma Membrane Transport Proteins: A group of membrane transport proteins that transport biogenic amine derivatives of catechol across the PLASMA MEMBRANE. Catecholamine plasma membrane transporter proteins regulate neural transmission as well as catecholamine metabolism and recycling.Carboxypeptidase H: A ZINC-containing exopeptidase primarily found in SECRETORY VESICLES of endocrine and neuroendocrine cells. It catalyzes the cleavage of C-terminal ARGININE or LYSINE residues from polypeptides and is active in processing precursors of PEPTIDE HORMONES and other bioactive peptides.Enkephalins: One of the three major families of endogenous opioid peptides. The enkephalins are pentapeptides that are widespread in the central and peripheral nervous systems and in the adrenal medulla.Secretory Vesicles: Vesicles derived from the GOLGI APPARATUS containing material to be released at the cell surface.Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone: A proton ionophore that is commonly used as an uncoupling agent in biochemical studies.Reserpine: An alkaloid found in the roots of Rauwolfia serpentina and R. vomitoria. Reserpine inhibits the uptake of norepinephrine into storage vesicles resulting in depletion of catecholamines and serotonin from central and peripheral axon terminals. It has been used as an antihypertensive and an antipsychotic as well as a research tool, but its adverse effects limit its clinical use.Cell Fractionation: Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS.Enkephalin, Methionine: One of the endogenous pentapeptides with morphine-like activity. It differs from LEU-ENKEPHALIN by the amino acid METHIONINE in position 5. Its first four amino acid sequence is identical to the tetrapeptide sequence at the N-terminal of BETA-ENDORPHIN.Phenylethanolamine N-Methyltransferase: A methyltransferase that catalyzes the reaction of S-adenosyl-L-methionine and phenylethanolamine to yield S-adenosyl-L-homocysteine and N-methylphenylethanolamine. It can act on various phenylethanolamines and converts norepinephrine into epinephrine. (From Enzyme Nomenclature, 1992) EC 2.1.1.28.Membranes: Thin layers of tissue which cover parts of the body, separate adjacent cavities, or connect adjacent structures.Norepinephrine: Precursor of epinephrine that is secreted by the adrenal medulla and is a widespread central and autonomic neurotransmitter. Norepinephrine is the principal transmitter of most postganglionic sympathetic fibers and of the diffuse projection system in the brain arising from the locus ceruleus. It is also found in plants and is used pharmacologically as a sympathomimetic.Dicyclohexylcarbodiimide: A carbodiimide that is used as a chemical intermediate and coupling agent in peptide synthesis. (From Hawley's Condensed Chemical Dictionary, 12th ed)Vesicular Biogenic Amine Transport Proteins: Integral membrane proteins of the LIPID BILAYER of SECRETORY VESICLES that catalyze transport and storage of biogenic amine NEUROTRANSMITTERS such as ACETYLCHOLINE; SEROTONIN; MELATONIN; HISTAMINE; and CATECHOLAMINES. The transporters exchange vesicular protons for cytoplasmic neurotransmitters.Proton-Translocating ATPases: Multisubunit enzymes that reversibly synthesize ADENOSINE TRIPHOSPHATE. They are coupled to the transport of protons across a membrane.Kinetics: The rate dynamics in chemical or physical systems.Calcium: A basic element found in nearly all organized tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.Adenosine Triphosphate: An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.Nigericin: A polyether antibiotic which affects ion transport and ATPase activity in mitochondria. It is produced by Streptomyces hygroscopicus. (From Merck Index, 11th ed)Microscopy, Electron: Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.Epinephrine: The active sympathomimetic hormone from the ADRENAL MEDULLA. It stimulates both the alpha- and beta- adrenergic systems, causes systemic VASOCONSTRICTION and gastrointestinal relaxation, stimulates the HEART, and dilates BRONCHI and cerebral vessels. It is used in ASTHMA and CARDIAC FAILURE and to delay absorption of local ANESTHETICS.Ascorbic Acid: A six carbon compound related to glucose. It is found naturally in citrus fruits and many vegetables. Ascorbic acid is an essential nutrient in human diets, and necessary to maintain connective tissue and bone. Its biologically active form, vitamin C, functions as a reducing agent and coenzyme in several metabolic pathways. Vitamin C is considered an antioxidant.Vesicular Monoamine Transport Proteins: A family of vesicular amine transporter proteins that catalyze the transport and storage of CATECHOLAMINES and indolamines into SECRETORY VESICLES.Tyramine: An indirect sympathomimetic. Tyramine does not directly activate adrenergic receptors, but it can serve as a substrate for adrenergic uptake systems and monoamine oxidase so it prolongs the actions of adrenergic transmitters. It also provokes transmitter release from adrenergic terminals. Tyramine may be a neurotransmitter in some invertebrate nervous systems.Ethylmaleimide: A sulfhydryl reagent that is widely used in experimental biochemical studies.Dienestrol: A synthetic, non-steroidal estrogen structurally related to stilbestrol. It is used, usually as the cream, in the treatment of menopausal and postmenopausal symptoms.Hexestrol: A synthetic estrogen that has been used as a hormonal antineoplastic agent.Cytochrome b Group: Cytochromes (electron-transporting proteins) with protoheme (HEME B) as the prosthetic group.Hydrogen-Ion Concentration: The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH = log 1/2[1/(H+)], where (H+) is the hydrogen ion concentration in gram equivalents per liter of solution. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)Annexins: Family of calcium- and phospholipid-binding proteins which are structurally related and exhibit immunological cross-reactivity. Each member contains four homologous 70-kDa repeats. The annexins are differentially distributed in vertebrate tissues (and lower eukaryotes) and appear to be involved in MEMBRANE FUSION and SIGNAL TRANSDUCTION.Cells, Cultured: Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.Centrifugation, Density Gradient: Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Ca(2+) Mg(2+)-ATPaseProprotein Convertase 2: A serine endopeptidase that has specificity for cleavage at ARGININE. It cleaves a variety of prohormones including PRO-OPIOMELANOCORTIN, proluteinizing-hormone-releasing hormone, proenkephalins, prodynorphin, and PROINSULIN.Nerve Tissue ProteinsMethylaminesCell Membrane: The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.Syntaxin 1: A neuronal cell membrane protein that combines with SNAP-25 and SYNAPTOBREVIN 2 to form a SNARE complex that leads to EXOCYTOSIS.Tyrosine 3-Monooxygenase: An enzyme that catalyzes the conversion of L-tyrosine, tetrahydrobiopterin, and oxygen to 3,4-dihydroxy-L-phenylalanine, dihydrobiopterin, and water. EC 1.14.16.2.Valinomycin: A cyclododecadepsipeptide ionophore antibiotic produced by Streptomyces fulvissimus and related to the enniatins. It is composed of 3 moles each of L-valine, D-alpha-hydroxyisovaleric acid, D-valine, and L-lactic acid linked alternately to form a 36-membered ring. (From Merck Index, 11th ed) Valinomycin is a potassium selective ionophore and is commonly used as a tool in biochemical studies.Microscopy, Immunoelectron: Microscopy in which the samples are first stained immunocytochemically and then examined using an electron microscope. Immunoelectron microscopy is used extensively in diagnostic virology as part of very sensitive immunoassays.Molecular Weight: The sum of the weight of all the atoms in a molecule.Amino Acid Sequence: The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.Adenosine Triphosphatases: A group of enzymes which catalyze the hydrolysis of ATP. The hydrolysis reaction is usually coupled with another function such as transporting Ca(2+) across a membrane. These enzymes may be dependent on Ca(2+), Mg(2+), anions, H+, or DNA.Membrane Fusion: The adherence and merging of cell membranes, intracellular membranes, or artificial membranes to each other or to viruses, parasites, or interstitial particles through a variety of chemical and physical processes.Thiocyanates: Organic derivatives of thiocyanic acid which contain the general formula R-SCN.Subcellular Fractions: Components of a cell produced by various separation techniques which, though they disrupt the delicate anatomy of a cell, preserve the structure and physiology of its functioning constituents for biochemical and ultrastructural analysis. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p163)Biological Transport: The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments.Carboxypeptidases: Enzymes that act at a free C-terminus of a polypeptide to liberate a single amino acid residue.Molecular Sequence Data: Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.Veratridine: A benzoate-cevane found in VERATRUM and Schoenocaulon. It activates SODIUM CHANNELS to stay open longer than normal.Endorphins: One of the three major groups of endogenous opioid peptides. They are large peptides derived from the PRO-OPIOMELANOCORTIN precursor. The known members of this group are alpha-, beta-, and gamma-endorphin. The term endorphin is also sometimes used to refer to all opioid peptides, but the narrower sense is used here; OPIOID PEPTIDES is used for the broader group.Membrane Proteins: Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.Vesicular Transport Proteins: A broad category of proteins involved in the formation, transport and dissolution of TRANSPORT VESICLES. They play a role in the intracellular transport of molecules contained within membrane vesicles. Vesicular transport proteins are distinguished from MEMBRANE TRANSPORT PROTEINS, which move molecules across membranes, by the mode in which the molecules are transported.Magnesium: A metallic element that has the atomic symbol Mg, atomic number 12, and atomic weight 24.31. It is important for the activity of many enzymes, especially those involved in OXIDATIVE PHOSPHORYLATION.Dopamine: One of the catecholamine NEUROTRANSMITTERS in the brain. It is derived from TYROSINE and is the precursor to NOREPINEPHRINE and EPINEPHRINE. Dopamine is a major transmitter in the extrapyramidal system of the brain, and important in regulating movement. A family of receptors (RECEPTORS, DOPAMINE) mediate its action.Neuropeptides: Peptides released by NEURONS as intercellular messengers. Many neuropeptides are also hormones released by non-neuronal cells.Membrane Potentials: The voltage differences across a membrane. For cellular membranes they are computed by subtracting the voltage measured outside the membrane from the voltage measured inside the membrane. They result from differences of inside versus outside concentration of potassium, sodium, chloride, and other ions across cells' or ORGANELLES membranes. For excitable cells, the resting membrane potentials range between -30 and -100 millivolts. Physical, chemical, or electrical stimuli can make a membrane potential more negative (hyperpolarization), or less negative (depolarization).Amines: A group of compounds derived from ammonia by substituting organic radicals for the hydrogens. (From Grant & Hackh's Chemical Dictionary, 5th ed)Digitonin: A glycoside obtained from Digitalis purpurea; the aglycone is digitogenin which is bound to five sugars. Digitonin solubilizes lipids, especially in membranes and is used as a tool in cellular biochemistry, and reagent for precipitating cholesterol. It has no cardiac effects.Vacuolar Proton-Translocating ATPases: Proton-translocating ATPases that are involved in acidification of a variety of intracellular compartments.Cerebellum: The part of brain that lies behind the BRAIN STEM in the posterior base of skull (CRANIAL FOSSA, POSTERIOR). It is also known as the "little brain" with convolutions similar to those of CEREBRAL CORTEX, inner white matter, and deep cerebellar nuclei. Its function is to coordinate voluntary movements, maintain balance, and learn motor skills.Potassium: An element in the alkali group of metals with an atomic symbol K, atomic number 19, and atomic weight 39.10. It is the chief cation in the intracellular fluid of muscle and other cells. Potassium ion is a strong electrolyte that plays a significant role in the regulation of fluid volume and maintenance of the WATER-ELECTROLYTE BALANCE.Protein Precursors

Recombinant SFD isoforms activate vacuolar proton pumps. (1/429)

The vacuolar proton pump of clathrin-coated vesicles is composed of two general sectors, a cytosolic, ATP hydrolytic domain (V1) and an intramembranous proton channel, V0. V1 is comprised of 8-9 subunits including polypeptides of 50 and 57 kDa, termed SFD (Sub Fifty-eight-kDa Doublet). Although SFD is essential to the activation of ATPase and proton pumping activities catalyzed by holoenzyme, its constituent polypeptides have not been separated to determine their respective roles in ATPase functions. Recent molecular characterization of these subunits revealed that they are isoforms that arise through an alternative splicing mechanism (Zhou, Z., Peng, S.-B., Crider, B.P., Slaughter, C., Xie, X.S., and Stone, D.K. (1998) J. Biol. Chem. 273, 5878-5884). To determine the functional characteristics of the 57-kDa (SFDalpha)1 and 50-kDa (SFDbeta) isoforms, we expressed these proteins in Escherichia coli. We determined that purified recombinant proteins, rSFDalpha and rSFDbeta, when reassembled with SFD-depleted holoenzyme, are functionally interchangeable in restoration of ATPase and proton pumping activities. In addition, we determined that the V-pump of chromaffin granules has only the SFDalpha isoform in its native state and that rSFDalpha and rSFDbeta are equally effective in restoring ATPase and proton pumping activities to SFD-depleted enzyme. Finally, we found that SFDalpha and SFDbeta structurally interact not only with V1, but also withV0, indicating that these activator subunits may play both structural and functional roles in coupling ATP hydrolysis to proton flow.  (+info)

Early requirement for alpha-SNAP and NSF in the secretory cascade in chromaffin cells. (2/429)

NSF and alpha-SNAP have been shown to be required for SNARE complex disassembly and exocytosis. However, the exact requirement for NSF and alpha-SNAP in vesicular traffic through the secretory pathway remains controversial. We performed a study on the kinetics of exocytosis from bovine chromaffin cells using high time resolution capacitance measurement and electrochemical amperometry, combined with flash photolysis of caged Ca2+ as a fast stimulus. alpha-SNAP, a C-terminal mutant of alpha-SNAP, and NEM were assayed for their effects on secretion kinetics. Two kinetically distinct components of catecholamine release can be observed upon fast step-like elevation of [Ca2+]i. One is the exocytotic burst, thought to represent the readily releasable pool of vesicles. Following the exocytotic burst, secretion proceeds slowly at maintained high [Ca2+]i, which may represent vesicle maturation/recruitment, i.e. some priming steps after docking. alpha-SNAP increased the amplitude of both the exocytotic burst and the slow component but did not change their kinetics, which we examined with millisecond time resolution. In addition, NEM only partially inhibited the slow component without altering the exocytotic burst, fusion kinetics and the rate of endocytosis. These results suggest a role for alpha-SNAP/NSF in priming granules for release at an early step, but not modifying the fusion of readily releasable granules.  (+info)

Sympathomimetic effects of MIBG: comparison with tyramine. (3/429)

Because nothing is known about whether metaiodobenzylguanidine (MIBG) has tyramine-like actions, the sympathomimetic effects of MIBG were determined in the isolated rabbit heart and compared with those of tyramine. METHODS: Spontaneously beating rabbit hearts were perfused with Tyrode's solution (Langendorff technique; 37 degrees C; 26 mL/min), and the heart rate as well as the norepinephrine and dopamine overflow into the perfusate was measured before and after doses of MIBG or tyramine (0.03-10 micromol) given as bolus injections (100 microL) into the aortic cannula. Km and Vmax values for the neuronal uptake (uptake1) of 125I-MIBG and 14C-tyramine were obtained in human neuroblastoma (SK-N-SH) cells. The Ki of MIBG for inhibition of the 3H-catecholamine uptake mediated by the vesicular monoamine transporter was determined in membrane vesicles obtained from bovine chromaffin granules and compared with the previously reported Ki value for tyramine determined under identical experimental conditions. RESULTS: By producing increases in heart rate and norepinephrine overflow, both compounds had dose-dependent sympathomimetic effects in the rabbit heart. MIBG was much less effective than tyramine in increasing heart rate (maximum effect 59 versus 156 beats/min) and norepinephrine overflow (maximum effect 35 versus 218 pmol/g). Tyramine also caused increases in dopamine overflow, whereas MIBG was a poor dopamine releaser. At a dose of 10 micromol, the increase in heart rate lasted more than 60 min after MIBG and about 20 min after tyramine injection. Accordingly, the norepinephrine overflow caused by 10 micromol MIBG and tyramine declined with half-lives of 57.8 and 2.2 min, respectively. The effects of both drugs were drastically reduced in hearts exposed to 2 micromol/L desipramine. The kinetic parameters characterizing the saturation of neuronal uptake by 125I-MIBG and 14C-tyramine were similar for the two compounds: Km values of MIBG and tyramine were 1.6 and 1.7 micromol/L, respectively, and Vmax values of MIBG and tyramine were 43 and 37 pmol/mg protein/min, respectively. However, in inhibiting the vesicular 3H-catecholamine uptake, MIBG was eight times less potent than tyramine. CONCLUSION: MIBG is much less effective than tyramine as an indirect sympathomimetic agent. This is probably a result of its relatively low affinity for the vesicular monoamine transporter and explains the relatively poor ability of the drug to mobilize norepinephrine stored in synaptic vesicles. The long duration of MIBG action results primarily from the drug not being metabolized by monoamine oxidase. The sympathomimetic effects of MIBG described here are not likely to come into play in patients given diagnostic or common therapeutic doses of radioiodinated MIBG.  (+info)

nSec-1 (munc-18) interacts with both primed and unprimed syntaxin 1A and associates in a dimeric complex on adrenal chromaffin granules. (4/429)

The target-SNARE syntaxin 1A is an essential component of the core machinery required for regulated exocytosis (where SNARE is the soluble N-ethylmaleimide-sensitive fusion protein-attachment protein receptor). Syntaxin 1A interacts with a variety of other proteins, two of which, N-ethylmaleimide-sensitive fusion protein (NSF) and alpha-soluble NSF attachment protein (alpha-SNAP) have been suggested to impart a conformational rearrangement on this protein during a reaction referred to as priming. We have studied the effect of the primed state on the binding properties of syntaxin 1A and we have confirmed that primed syntaxin 1A no longer associated with alpha-SNAP or its cognate vesicle-SNARE, vesicle-associated membrane protein (VAMP). Under such conditions, however, it retained the ability to bind to nSec-1. It has been demonstrated that nSec-1, a regulatory protein also involved in neuronal exocytosis, binds syntaxin 1A with high affinity in vitro, although evidence for this physical interaction occurring in vivo has proven elusive. We analysed the subcellular distribution of these two proteins in fractions from bovine adrenal medulla and detected syntaxin 1A and nSec-1 in both plasma membrane and chromaffin-granule fractions. Using a cross-linking approach with chromaffin-granule membranes we detected a putative dimeric complex composed of approx. 54% total granule membrane nSec-1 and approx. 30% total syntaxin 1A. The results of this study therefore suggest the possibility of nSec-1 interactions with primed syntaxin 1A and demonstrate a potentially significant interaction of syntaxin 1A and nSec-1 on the membranes of chromaffin granules.  (+info)

Molecular cloning of endopin 1, a novel serpin localized to neurosecretory vesicles of chromaffin cells. Inhibition of basic residue-cleaving proteases by endopin 1. (5/429)

Serpins represent a diverse class of endogenous protease inhibitors that regulate important biological functions. In consideration of the importance of regulated proteolysis within secretory vesicles for the production of peptide hormones and neurotransmitters, this study revealed the molecular identity of a novel serpin, endopin 1, that is localized to neurosecretory vesicles of neuropeptide-containing chromaffin cells (chromaffin granules). Endopin 1 of 68-70 kDa was present within isolated chromaffin granules. Stimulated cosecretion of endopin 1 with chromaffin granule components, [Met]enkephalin and a cysteine protease known as "prohormone thiol protease," demonstrated localization of endopin 1 to functional secretory vesicles. Punctate, discrete immunofluorescence cellular localization of endopin 1 in chromaffin cells was consistent with its secretory vesicle localization. Endopin 1 contains a unique reactive site loop with Arg as the predicted P1 residue, suggesting inhibition of basic residue-cleaving proteases; indeed, trypsin was potently inhibited (K(i(app)) of 5 nM), and plasmin was moderately inhibited. Although endopin 1 possesses homology with alpha(1)-antichymotrypsin, chymotrypsin was not inhibited. Moreover, endopin 1 inhibited the chromaffin granule prohormone thiol protease (involved in proenkephalin processing). These results suggest a role for the novel serpin, endopin 1, in regulating basic residue-cleaving proteases within neurosecretory vesicles of chromaffin cells.  (+info)

Comparison of cysteine string protein (Csp) and mutant alpha-SNAP overexpression reveals a role for csp in late steps of membrane fusion in dense-core granule exocytosis in adrenal chromaffin cells. (6/429)

Assembly of the SNARE complex and its disassembly caused by the action of soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein (SNAP) and NSF is crucial for the maintenance of vesicular traffic, including fusion of regulated exocytotic vesicles. Various other proteins may also have important roles in the processes leading to membrane fusion via interaction with the SNARE proteins, including the secretory vesicle cysteine string protein (Csp). Here we have examined the effect of overexpression of a dominant negative alpha-SNAP mutant or Csp on exocytosis of dense-core granules in single chromaffin cells monitored using amperometry to detect released catecholamine. Exocytosis of trans-Golgi network (TGN)-derived dense-core granules was substantially inhibited by expression of alpha-SNAP(L294A). The amplitude and characteristics of the individual release events were unaffected by expression of alpha-SNAP(L294A), consistent with an essential role for alpha-SNAP in early steps of priming but not in the fusion process. In contrast, Csp overexpression, which also inhibited the extent of exocytosis, also modified the kinetics of the individual release events seen as an increase in the rise time and a broadening of the residual amperometric spikes in Csp-transfected cells. These results suggest that unlike alpha-SNAP, Csp plays a key role in the protein interactions close to the fusion process or fusion pore opening during Ca(2+)-regulated exocytosis.  (+info)

A pleckstrin homology domain specific for phosphatidylinositol 4, 5-bisphosphate (PtdIns-4,5-P2) and fused to green fluorescent protein identifies plasma membrane PtdIns-4,5-P2 as being important in exocytosis. (7/429)

Kinetically distinct steps can be distinguished in the secretory response from neuroendocrine cells with slow ATP-dependent priming steps preceding the triggering of exocytosis by Ca(2+). One of these priming steps involves the maintenance of phosphatidylinositol 4, 5-bisphosphate (PtdIns-4,5-P(2)) through lipid kinases and is responsible for at least 70% of the ATP-dependent secretion observed in digitonin-permeabilized chromaffin cells. PtdIns-4,5-P(2) is usually thought to reside on the plasma membrane. However, because phosphatidylinositol 4-kinase is an integral chromaffin granule membrane protein, PtdIns-4,5-P(2) important in exocytosis may reside on the chromaffin granule membrane. In the present study we have investigated the localization of PtdIns-4,5-P(2) that is involved in exocytosis by transiently expressing in chromaffin cells a pleckstrin homology (PH) domain that specifically binds PtdIns-4, 5-P(2) and is fused to green fluorescent protein (GFP). The PH-GFP protein predominantly associated with the plasma membrane in chromaffin cells without any detectable association with chromaffin granules. Rhodamine-neomycin, which also binds to PtdIns-4,5-P(2), showed a similar subcellular localization. The transiently expressed PH-GFP inhibited exocytosis as measured by both biochemical and electrophysiological techniques. The results indicate that the inhibition was at a step after Ca(2+) entry and suggest that plasma membrane PtdIns-4,5-P(2) is important for exocytosis. Expression of PH-GFP also reduced calcium currents, raising the possibility that PtdIns-4,5-P(2) in some manner alters calcium channel function in chromaffin cells.  (+info)

Nitric oxide modulates a late step of exocytosis. (8/429)

The effects of nitric oxide (NO) on the late phase of exocytosis have been studied, by amperometry, on Ba(2+)-stimulated chromaffin cells. Acute incubation with NO or NO donors (sodium nitroprusside, spermine-NO, S-nitrosoglutathione) produced a drastic slowdown of the granule emptying. Conversely, cell treatment with N(omega)-nitro-l-arginine methyl ester (a NO synthase inhibitor) or with NO scavengers (methylene blue, 2-(4-carboxyphenyl)-4,4,5, 5-tetramethyl-imidazoline-1-oxyl-3-oxide potassium) accelerated the extrusion of catecholamines from chromaffin granules, suggesting the presence of a NO modulatory tone. The incubation with phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine or zaprinast) or with the cell-permeant cGMP analog 8-bromo-cGMP, mimicked the effects of NO, suggesting the involvement of the guanylate cyclase cascade. NO effects were not related to changes in intracellular Ba(2+). NO did not modify the duration of feet. Effects were evident even on pre-fusioned granules, observed under hypertonic conditions, suggesting that the fusion pore is not the target for NO, which probably acts by modifying the affinity of catecholamines for the intragranular matrix. NO could modify the synaptic transmitter efficacy through a novel mechanism, which involves the regulation of the emptying of secretory vesicles.  (+info)

*V-ATPase

Johnson RG, Beers MF, Scarpa A; Beers; Scarpa (September 1982). "H+ ATPase of chromaffin granules. Kinetics, regulation, and ...

*ATPase

Njus, D.; Knoth, J.; Zallakian, M. (1981). "Proton-linked transport in chromaffin granules". Curr. Top. Bioenerg. 11: 107-147. ...

*Ascorbate ferrireductase (transmembrane)

Flatmark, T.; Terland, O. (1971). "Cytochrome b561 of the bovine adrenal chromaffin granules. A high potential b-type ...

*ATP6V0D1

An accessory polypeptide of the H+-ATPase from chromaffin granules". J. Biol. Chem. 263 (33): 17638-42. PMID 2903164. Bonaldo ...

*Morpheein

Stewart, L C; Klinman, J P (1988). "Dopamine Beta-Hydroxylase of Adrenal Chromaffin Granules: Structure and Function". Annual ...

*CYB561

Duong LT, Fleming PJ (1982). "Isolation and properties of cytochrome b561 from bovine adrenal chromaffin granules". J. Biol. ... an integral protein of the chromaffin granule membrane". J. Biol. Chem. 270 (39): 22714-20. doi:10.1074/jbc.270.39.22714. PMID ...

*Chromophil

... cells are mostly hormone-producing cells containing so-called chromaffin granules. In these subcellular structures, ...

*Chromaffin cell

The hormones are secreted from chromaffin granules; this is where the enzyme dopamine β-hydroxylase catalyses the conversion of ... in Adrenal Chromaffin Cells UC-San Diego Chromaffin Cell and Hypertension Research A Primer on Chromaffin Cells Rat Chromaffin ... Szewczyk, A; Lobanov, NA; Kicińska, A; Wójcik, G; Nałecz, MJ (2001). "ATP-sensitive K+ transport in adrenal chromaffin granules ... Chromaffin cells also settle near the vagus nerve and carotid arteries. In lower concentrations, extra-adrenal chromaffin cells ...

*Vesicular monoamine transporter

VMAT was first isolated and purified in bovine chromaffin granules, in both native and denatured forms. There are two types of ... It is thought that chromaffin granules possess these heterotrimeric G-proteins which have shown to be regulatory to small clear ... VMAT1 may be found in neuroendocrine cells, particularly chromaffin and enterochromaffin granules which are largely found in ... 1994). "Biochemistry and molecular biology of the vesicular monoamine transporter from chromaffin granules". J Exp Biol. 196: ...

*Chromaffin

Chromophil cells, hormone producing cells showing chromaffin granules that readily absorb chromium stains.. ... Chromaffin may refer to: Chromaffin cells, neuroendocrine cells in the adrenal medulla. ...

*Carboxypeptidase E

... and adrenal gland chromaffin cells. Within cells, carboxypeptidase E is present in the secretory granules along with its ... and characterization of a specific enkephalin-synthesizing carboxypeptidase localized to adrenal chromaffin granules". ... acts as a sorting signal for regulated secretory proteins in the trans-Golgi network of the pituitary and in secretory granules ...

*ATP6V1C1

1990). "Molecular cloning of cDNA encoding the C subunit of H(+)-ATPase from bovine chromaffin granules". J. Biol. Chem. 265 ( ...

*Chromogranin A

... of glycosylated and phosphorylated chromogranin A-derived peptide 173-194 from bovine adrenal medullary chromaffin granules". ... It is present in islet beta cell secretory granules. Chromogranin A is the precursor to several functional peptides including ... Examples of cells producing chromogranin A (ChgA) are chromaffin cells of the adrenal medulla, paraganglia, enterochromaffin- ... it is located in secretory vesicles of neurons and endocrine cells such as islet beta cell secretory granules in pancreas. In ...

*Annexin

... that causes calcium-dependent aggregation of isolated chromaffin granules". Journal of Biological Chemistry. 253 (8): 2858-66. ... was the first annexin to be discovered while searching for proteins that promote the contact and fusion of chromaffin granules ... authors used bovine adrenal glands and identified a calcium dependent protein that was responsible for aggregation of granules ...

*Nicotine

Calcium triggers the exocytosis of chromaffin granules and thus the release of epinephrine (and norepinephrine) into the ...

*ATP6V0E1

... and characterization of a novel 9.2-kDa membrane sector-associated protein of vacuolar proton-ATPase from chromaffin granules ...

*Biological membrane

... and chromaffin granules). Different types of biological membranes have diverse lipid and protein compositions. The content of ... cytoplasmic granules; cell vesicles (phagosome, autophagosome, clathrin-coated vesicles, COPI-coated and COPII-coated vesicles ...

*List of MeSH codes (A11)

... cytoplasmic granules MeSH A11.284.430.214.190.875.190.190.207 - chromaffin granules MeSH A11.284.430.214.190.875.190.190.560 - ... cytoplasmic granules MeSH A11.284.430.214.190.500.207 - chromaffin granules MeSH A11.284.430.214.190.500.560 - melanosomes MeSH ...

*List of MeSH codes (A06)

File "2006 MeSH Trees".) MeSH A06.224.161 --- chromaffin cells MeSH A06.224.207 --- chromaffin granules MeSH A06.224.358 --- ... chromaffin MeSH A06.390.021 --- enterochromaffin cells MeSH A06.390.043 --- enterochromaffin-like cells MeSH A06.390.065 --- ...

*Shimon Schuldiner

Identification and Purification of a Functional Amine Transporter from Bovine Chromaffin Granules. J. Biol. Chem. 265, 3961- ...

*MARCKS

Recently, MARCKS has been implicated in the exocytosis of a number of vesicles and granules such as mucin and chromaffin. It is ...

*Adrenal gland

Cells of the adrenal medulla are called chromaffin cells because they contain granules that stain with chromium salts, a ... The chromaffin cells of the medulla are the body's main source of the catecholamines adrenaline and noradrenaline, released by ... Formation Catecholamines are produced in chromaffin cells in the medulla of the adrenal gland, from tyrosine, a non-essential ... Pheochromocytomas are tumors of the adrenal medulla that arise from chromaffin cells. They can produce a variety of nonspecific ...

*Granule

... spherical granules of material found on the surface of the planet Mars Granule (geology), a specified particle size of 2-4 ... a structure characteristic of the azurophil eukarytotic cell type Chromaffin granule, a structure characteristic of the ...

*Paraganglioma

While all contain neurosecretory granules, only in 1-3% of cases is secretion of hormones such as catecholamines abundant ... which however are chromaffin-positive. Most paragangliomas are either asymptomatic or present as a painless mass. ... comprising a small mass of neural crest-derived chromaffin cells. Serves as a common origin of abdominal paragangliomas. Glomus ... Paragangliomas originate from paraganglia in chromaffin-negative glomus cells derived from the embryonic neural crest, ...

*Granule (cell biology)

The granules of leukocytes are classified as azurophilic granules or specific granules. Leukocyte granules are released in ... Chromaffin granule Kurloff cell "granule" at Dorland's Medical Dictionary André J, Rouiller CH (1957) L'ultrastructure de la ... The granules of platelets are classified as dense granules and alpha granules. In 1957, André and Rouiller first coined the ... Insulin granules are secretory granules, which can release their contents from the cell into the bloodstream. The beta cells in ...

*Enterochromaffin cell

Ultrastructurally, these granules are reported to vary in size and shape and are considered pleomorphic. Most EC cells ... Similarly named, Chromaffin cells (of the adrenal medulla) share this characteristic and are histologically similar to EC cells ... The name 'enterochromaffin' comes from the Greek word "enteron" (ἔντερον), in relation to intestines, and "chromaffin" as a ...
Membranes of chromaffin granules were isolated from the adrenal glands of four different species. The solubilized membrane proteins could be resolved into several bands by polyacrylamide-gel electrophoresis (alkaline and acid gel systems). Two major protein components appeared to be common to the chromaffin granule membranes of ox, horse, pig and man. The various membrane proteins of bovine chromaffin granules were separated by filtration on Sephadex G-200 in the presence of sodium dodecyl sulphate. Two major membrane proteins (A and B) were obtained in purified form. Treatment of protein A with 2-mercaptoethanol before electrophoresis resulted in two more rapidly migrating subunits, whereas protein B was unaffected by mercaptoethanol treatment. The amino acid compositions of the two purified proteins were determined. They are very similar to that of the total membrane proteins but significantly different from that of the chromogranins, the soluble proteins of chromaffin granules.. ...
Dopamine-ß-hydroxylase (DßH), an enzyme which catalyzes the conversion of dopamine to norepinephrine, is the only enzyme of the catecholamine biosynthetic pathway located in the chromaffin granules of adrenal medulla. Within the granules, two populations of DßH exist: a water-soluble fraction found within the granule matrix and a membrane-bound, amphiphilic fraction embedded in the surrounding bilayer. The amphiphilic form was purified to homogeneity following its extraction from the membrane with the non-ionic detergent BRIJ 58. Three steps were required to achieve complete purification: adsorption to ConA-Sepharose, adsorption to DEAE Sephadex A-25, and chromatography on Sephacryl S-200, Sepharose 6B, or Sepharose CL-4B. The presence of 0.1-0.2 mg/ml BRIJ 58 was essential for protein recovery. The enzymatic and structural characteristics of membrane-bound DßH were found to be similar to those of soluble DßH. Initial velocity data indicated a Ping-pong or double-displacement reaction with ...
The bovine homologue of p65, a calmodulin-binding protein located in the membranes of synaptic vesicles and endocrine secretory granules, has been studied by the use of monoclonal antibodies directed against this antigen and against dopamine beta-mono-oxygenase. The protein (apparent molecular mass 67 kDa; pI = 5.5-6.2) is partially degraded by treatment with neuraminidase or endoglycosidase F. Trypsin treatment of intact adrenal chromaffin granules or of granule membranes releases a soluble 39 kDa fragment of p65 which corresponds to the whole of its cytoplasmic domain. This domain contains both the epitope for the monoclonal antibody cgm67 and the calmodulin-binding site. The 20 amino acids at the N-terminus of this fragment are identical to part of the rat p65 sequence.. ...
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
Pomegranate (Punica granatum) is an edible fruit originating in the Middle East, the juice of which is widely available commercially. Various parts of the fruit including the juice have been used traditionally to treat a range of ailments.. The juice and other extracts have shown a wide range of bioactivity in pre-clinical studies, such as anti-inflammatory, anti-infective and anti-oxidant effects.. On the basis of this research, beneficial effects are claimed in cancer, specifically in the prevention and treatment of prostate cancer.. Few clinical trials in cancer have been conducted to date: an uncontrolled trial showed promising beneficial effects on prostate specific antigen (PSA) doubling time in prostate cancer but subsequent randomised controlled trials have failed to show any difference between pomegranate and placebo. One small randomized clinical trial did not show an effect of pomegranate consumption on breast cancer risk.. Pomegranate juice has been widely consumed for many years. It ...
Cultures of bovine adrenomedullary chromaffin cells accumulated 1-methyl-4-phenylpyridinium (MPP+) in a time- and concentration-dependent manner by a process that was prevented by desmethylimipramine. The subcellular localization of the incorporated [methyl-3H]MPP+ was examined by differential centrifugation and sucrose density gradient fractionation and was found to be predominantly colocalized with catecholamines in chromaffin vesicles, and negligible amounts were detected within the mitochondrial fraction. When chromaffin cell membranes were made permeable with the detergent digitonin in the absence of calcium, there was no increase in the release of [3H]MPP+, indicating that there is negligible accumulation of the neurotoxin in the cytosol. Simultaneous exposure to digitonin and calcium induced cosecretion of MPP+ and catecholamines. Stimulation of the cells with nicotine released both catecholamines and MPP+ at identical rates and percentages of cellular content in a calcium-dependent ...
In addition to the canonical ribonucleoside and deoxyribonucleoside phosphates and cofactors, cells contain a large number of minor nucleotides. Among these are the diadenosine polyphosphates (ApnA, where n = 2-7 [1]). Ap3A and Ap4A are the most intensively studied of these and are generally present in the soluble fraction of eukaryotic and prokaryotic cells at concentrations between 10 nM and 5 μM [2]. Platelet dense granules, adrenal chromaffin granules and certain synaptic vesicles have been reported to contain high concentrations of Ap5A and Ap6A in addition to Ap3A and Ap4A, all of which can be exocytosed following appropriate stimuli and bind to target cell purinoceptors causing a variety of physiological responses in the cardiovascular and central and peripheral nervous systems [1, 3-5]. However, although Ap6A has been detected in erythrocytes [6], there are no substantiated measurements of Ap5A and Ap6A in the soluble fraction of nucleated cells, and it is likely that they are typically ...
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Dopamine beta-monooxygenase is shown to catalyze the oxidation of N,N,N,N-tetramethyl-1,4-phenylenediamine (TMPD) to its cation radical in the presence of a regular substrate and molecular oxygen. The enzyme-mediated oxidation of TMPD is stoichiometrically coupled with the hydoxylation of the substrate to the corresponding enzymatic product. TMPD is kinetically well behaved as an alternate electron donor for the enzyme with a potency comparable to that of the most efficient electron donor, ascorbate. Dopamine beta-monooxygenase mediated oxidation of TMPD has been employed to design a convenient and sensitive spectrophotometric assay for the enzyme. The finding that TMPD is a well behaved facile alternate electron donor for dopamine beta-monooxygenase raises some interesting novel questions regarding the specificity and chemistry of the reduction site, which may have important implications on the reduction of active site coppers of the enzyme ...
TY - JOUR. T1 - Microsequencing of dopamine beta-monooxygenase. AU - McCafferty, B.. AU - Angeletti, R. H.. PY - 1987. Y1 - 1987. N2 - Tryptic peptides and cyanogen bromide fragments of dopamine beta-monooxygenase (DBH) were prepared and separated on C-8 reverse phase columns by high pressure liquid chromatography. Absorbance profiles at both 220 nm and 280 nm were monitored so that peptides with aromatic residues could be isolated. These peptides were subjected to automated Edmann degradation with a gas phase microsequencer.. AB - Tryptic peptides and cyanogen bromide fragments of dopamine beta-monooxygenase (DBH) were prepared and separated on C-8 reverse phase columns by high pressure liquid chromatography. Absorbance profiles at both 220 nm and 280 nm were monitored so that peptides with aromatic residues could be isolated. These peptides were subjected to automated Edmann degradation with a gas phase microsequencer.. UR - ...
Rat pheochromocytoma cells (PC 12) permeabilized with staphylococcal α-toxin release [3H]dopamine after addition of micromolar Ca2+. This does not require additional Mg2+-ATP (in contrast to bovine adrenal medullary chromaffin cells). We also observed Ca2+-dependent [3H]-dopamine release from digitonin-permeabilized PC 12 cells. Permeabilization with α-toxin or digitonin and stimulation of the cells were done consecutively to wash out endogenous Mg2+-ATP. During permeabilization, ATP was removed effectively from the cytoplasm by both agents but the cells released [3H]dopamine in response to micromolar Ca2+ alone. Replacement by chloride of glutamate, which could sustain mitochondrial ATP production in permeabilized cells, does not significantly alter catecholamine release induced by Ca2+. However, Mg2+ without ATP augments the Ca2+-induced release. The release was unaltered by thiol-, hydroxyl-, or calmodulin-interfering substances. Thus Mg2+-ATP, calmodulin, or proteins containing -SH or -OH ...
Definition of chromaffin cell in the Financial Dictionary - by Free online English dictionary and encyclopedia. What is chromaffin cell? Meaning of chromaffin cell as a finance term. What does chromaffin cell mean in finance?
Definition of Chromaffin cells in the Legal Dictionary - by Free online English dictionary and encyclopedia. What is Chromaffin cells? Meaning of Chromaffin cells as a legal term. What does Chromaffin cells mean in law?
Buy BAM (8-22) (Bovine Adrenal Medulla 8-22) (CAS 412961-36-5), a water soluble SNSR agonist. Join researchers using high quality BAM (8-22) (Bovine Adrenal…
DBH - Rabbit polyclonal antibody to Dopamine beta-Hydroxylase (dopamine beta-hydroxylase (dopamine beta-monooxygenase)) available from OriGene
Dopamine beta-hydroxylase (DBH; dopamine beta-monooxygenase) is a copper-containing glycoprotein consisting of four identical subunits and catalyzes the oxidation of dopamine to norepinephrine. It requires ascorbic acid as an electron donor. DBH is localized in the norepinephrinergic and epinephrinergic neurons in the central nervous system. The enzyme exists in the secretory vesicles as both soluble and membrane-bound forms. The soluble form is secreted with catecholamines by exocytosis whereas the membrane-bound form is recycled into the vesicles ...
Dopamine beta-hydroxylase (DBH; dopamine beta-monooxygenase) is a copper-containing glycoprotein consisting of four identical subunits and catalyzes the oxidation of dopamine to norepinephrine. It requires ascorbic acid as an electron donor. DBH is localized in the norepinephrinergic and epinephrinergic neurons in the central nervous system. The enzyme exists in the secretory vesicles as both soluble and membrane-bound forms. The soluble form is secreted with catecholamines by exocytosis whereas the membrane-bound form is recycled into the vesicles ...
Patient information for EPILIM CHRONOSPHERE 500MG MODIFIED RELEASE GRANULES Including dosage instructions and possible side effects.
Investigations into the effects of culturing bovine adrenal chromaffin cells in the presence (72 h) of dibutyryl cyclic AMP, forskolin, and reserpine on the level and release of [Met]enkephalyl-Arg6-Phe7 immunoreactivity, noradrenaline, and adrenaline are reported. The assay for [Met]enkephalyl-Arg6-Phe7 immunoreactivity recognises both peptide B, the 31-amino acid carboxy-terminal segment of proenkephalin, and its heptapeptide fragment, [Met]enkephalyl-Arg6-Phe7. Treatments that elevate cyclic AMP increase the amount of peptide immunoreactivity in these cells; this is predominantly peptide B-like immunoreactivity in both control cells and cyclic AMP-elevated cells. Treatment with reserpine gives no change in total immunoreactivity levels, but does not result in increased accumulation of the heptapeptide [Met]enkephalyl-Arg6-Phe7 at the expense of immunoreactivity that elutes with its immediate precursor, peptide B. Cyclic AMP treatment causes either no change or a decrease in levels of accumulated
TY - JOUR. T1 - Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells. AU - Perrais, David. AU - Kleppe, Ingo C.. AU - Taraska, Justin W.. AU - Almers, Wolfhard. PY - 2004/10/15. Y1 - 2004/10/15. N2 - After exocytosis, chromaffin granules release essentially all their catecholamines in small fractions of a second, but it is unknown how fast they release stored peptides and proteins. Here we compare the exocytic release of fluorescently labelled neuropeptide Y (NPY) and tissue plasminogen activator from single granules. Exocytosis was tracked by measuring the membrane capacitance, and single granules in live cells were imaged by evanescent field microscopy. Neuropeptide Y left most granules in small fractions of a second, while tissue plasminogen activator remained in open granules for minutes. Taking advantage of the dependence on pH of the fluorescence of green fluorescent protein, we used rhythmic external acidification to determine whether ...
Adrenal chromaffin cells (ACCs) secrete several neuroactive substances that are effective in influencing pain sensitivity in the central nervous system as well as enhancing the recovery of the intrinsic nigrostriatal dopaminergic system in patients w
The vesicular monoamine transporter acts to accumulate cytosolic monoamines into vesicles, using the proton gradient maintained across the vesicular membrane. Its proper function is essential to the correct activity of the monoaminergic systems that have been implicated in several human neuropsychiatric disorders. The transporter is a site of action of important drugs, including reserpine and tetrabenazine (Peter et al., 1993 [PubMed 7905859]). See also SLC18A2 (MIM 193001).[supplied by OMIM, Mar 2008 ...
In general, DA, after its synthesis, is taken up and stored in specialized subcellular organelles, the storage granules, to ensure its regulated release via exocytosis. Most information about the granular transport mechanism(s) and storage mechanism(s) of DA (and monoamines in general) has been obtained by using chromaffin granules isolated from adrenal medullary cells or PC12 cells as experimental substrate (Roda et al., 1980). Thus, it is presently known that the DAergic granular transport system consists of at least two components: (1) a so-called vesicular monoamine transporter (VMAT), structurally distinct from the plasma membrane DA transporter, and (2) a vacuolar-type ATP-driven H+ pump, which provides the electrochemical gradient on which the transporter depends for its function (Johnson, 1988; Schuldiner, 1994). Drugs such as reserpine and tetrabenazine deplete intracellular DA stores by selectively interfering with transmitter uptake via the VMAT. In the present set of experiments, we ...
A chromophil biological cell is a cell which is easily stainable by absorbing chromium salts used in histology to increase the visual contrast of samples for microscopy. Chromophil cells are mostly hormone-producing cells containing so-called chromaffin granules. In these subcellular structures, amino acid precursors to certain hormones are accumulated and subsequently decarboxylated to the corresponding amines, for example epinephrine, norepinephrine, dopamine or serotonin. Chromophil cells therefore belong to the group of APUD (amine precursor uptake and decarboxylation) cells. These cells are scattered throughout the whole body, but particularly in glands such as the hypothalamus, hypophysis, thyroid, parathyroid and pancreas ...
A process for making coated detergent granules by producing detergent granules and then uniformly distributing finely divided powder onto the surfaces of the detergent granules by means of an electrostatic charging and delivery system. The detergent granules are preferably glutinous (hot and/or sticky) and freely falling during application of the electrostatically charged powder particles. Detergent granules made according to this process are also covered.
TY - JOUR. T1 - Cloning and sequence analysis of cDNA for bovine carboxypeptidase E. AU - Fricker, Lloyd D.. AU - Evans, Chris J.. AU - Esch, Fred S.. AU - Herbert, Edward. PY - 1986/12/1. Y1 - 1986/12/1. N2 - Carboxypeptidase E (enkephalin convertase) was first identified as the carboxypeptidase B-like enzyme involved in the biosynthesis of enkephalin in bovine adrenal chromaffin granules1. A similar enzyme is present in many brain regions1,2 and in purified secretory granules from rat pituitary3 and rat insulinoma4. Within the secretory granules, carboxypeptidase E (CPE) activity is found in both a soluble and a membrane-bound form1, which differ slightly in relative molecular mass (Mr)5. Here, to investigate whether the CPE activities in the various tissues are produced from a single gene, purified CPE was partially sequenced and oligonucleotide probes were used to isolate a clone encoding CPE from a bovine pituitary complementary DNA library. This cDNA hybridizes to bovine pituitary poly(A)+ ...
TY - JOUR. T1 - Sodium-azide-evoked noradrenaline and catecholamine release from peripheral sympathetic nerves and chromaffin cells. AU - Török, Tamás L.. AU - Pauló, Tünde. AU - Tóth, Péter T.. AU - Azzidani, Awad M.. AU - Powis, David A.. AU - Magyar, K.. PY - 1989. Y1 - 1989. N2 - 1. 1. The spontaneous release of [3H]noradrenaline ([3H]NA) has been measured from rabbit pulmonary arteries and bovine chromaffin cells in the presence of neuronal uptake blocker cocaine (3 × 10-5 M). 2. 2. The Na+-pump inhibitor sodium-azide (NaN3, 2 mM) produced a moderate increase of [3H]NA release from both preparations and relaxed the arteries. The [3H]releasing action of NaN3 was accompanied by a 30% inhibition of 86Rb-uptake into chromaffin cells. 3. 3. In both preparations, ouabain (10-4 M) markedly increased the release of [3H], contracted the arteries and inhibited the 86Rb-uptake of chromaffin cells by about 75%. A combined application of NaN3 and ouabain produced a similar inhibition of ...
Common Functional Genetic Variants in Catecholamine Storage Vesicle Protein Promoter Motifs Interact to Trigger Systemic Hypertension Academic Article ...
Author: Nili, U. et al.; Genre: Journal Article; Published in Print: 2006-12-01; Title: Munc18-1 phosphorylation by protein kinase C potentiates vesicle pool replenishment in bovine chromaffin cells
Background: Hypertension is a complex trait, with deranged autonomic control of circulation. Chromogranin B (CHGB) is the most abundant core protein in human catecholamine secretory vesicles, playing an important role in their biogenesis. Does common interindividual variation at the CHGB locus contribute to phenotypic variation in CHGB and catecholamine secretion, autonomic stability of circulation, or blood pressure (BP) in the population? Methods and Results: To probe interindividual variability in CHGB, we systematically studied polymorphism across the locus by resequencing CHGB (≈6 kbp footprint spanning the promoter, 5 exons, exon/intron borders, untranslated regions) in 160 subjects (2n=320 chromosomes) of diverse biogeographic ancestries. We identified 53 single-nucleotide polymorphisms, of which 22 were common. We then studied 1182 subjects drawn from the most extreme BP values in the population (highest and lowest 5th percentiles), typing 4 common polymorphisms spanning the ≈14 kbp ...
The sympathetic nervous system is activated by a variety of threats to organismal homeostasis. The adrenomedullary chromaffin cell is the core effector of sympathetic activity in the peripheral nervous system. By design, the chromaffin cell secretory response is mutable so that release can be rapidly tuned to drive context-dependent changes in physiological function. However, the mechanisms by which this tuning is achieved with such high temporal fidelity and context specificity remain unclear. This represents a major gap in our understanding of the sympatho-adrenal system since it is known to modify the function of nearly every organ system in the body. In chromaffin cells, the trigger for stimulus-evoked exocytosis is a rise in intracellular Ca2+. The level of intracellular Ca2+ accumulation varies with the stimulus intensity and secretagogue. Ca2+ regulates release by acting on the Ca2+-binding synaptotagmin (Syt) protein family, driving their penetration into membranes that harbor anionic lipids,
A case-control study of 959 prevalent cases of parkinsonism (767 with PD) and 1989 controls across five European centres. Mucin gene mRNA levels in broilers challenged with eimeria and/or Clostridium perfringens. The bootstrapping procedure indicated that the parameter estimates were very stable, thus lending greater credence to the model.. These data provide a reference for the Y-STR database in Jilin Province, and they may be valuable for population genetic analysis. Ten-year experience using a plastic, disposable curette for the diagnosis of primary ciliary dyskinesia. Of the eight patients with Stage II-III disease, six patients eventually died of metastatic disease despite additional radiotherapy and/or chemotherapy.. These data provide important new insight into augmentin bambini the molecular mechanisms of apoptosis resistance in co-stimulated T cells. In chromaffin cells, ARF6 is specifically associated with the membrane of secretory chromaffin granules. Analysis of the complete ...
18 Adrenaline and noradrenaline: Most of the synthesis occurs in the adrenergic nerve ending and stored in granular vesicles called chromaffin granules close to the site of release into synaptic cleft . Biosynthesis could also occur in suprarenal medulla and other tissues. The enzyme (N-methyl transferase) which catalyses the conversion of noradrenaline to adrenaline occurs almost exclusively in suprarenal medulla and is therefore missing in the peripheral nerve terminals. Hence noradrenaline is the final step in the synthetic process in most adrenergic nerves. Catecholamines are sympathomimetics that contain the catechol nucleus (e.g. noradrenaline and adrenaline). Catecholamines are stored in synaptic granules in two forms in equilibrium: Bound noradrenaline with ATP and protein is the (inactive part). Free noradrenaline is released by nerve stimulation. Another portion of it is stored in the cytoplasm in free form (cytoplasmic free noradrenaline). ...
Synonyms for adrenomedullary hormones in Free Thesaurus. Antonyms for adrenomedullary hormones. 2 synonyms for hormone: endocrine, internal secretion. What are synonyms for adrenomedullary hormones?
Area of interest: Mechanisms of stress transduction at the sympatho-adrenal synapse; optical studies of hormone trafficking and secretion in the adrenomedullary chromaffin cell.
Shelly carbonate sands represent an extreme soil type in terms of their mechanical behavior which derives from the bioclastic nature of the constituent grains. In their uncemented form, these deposits exhibit very high compressibility, which has posed a number of geotechnical engineering problems; in most cases related to the reduction in the bearing capacities of both shallow and deep foundations. Remarkable features of these carbonate sands include the complex shape and the structural weakness of the grains and the high inter and intra granular porosity. Previous studies, have quoted the interlocking of the angular shelly particles to be at the origin of their high friction angles and high initial void ratio, however, up until now, no scientific micro-scale examination has been carried out. This paper presents a non-invasive image based investigation into the grain morphology of a carbonate sand from the Persian Gulf. This sand has a median grain size of 570μm and a high CaCO3 content in the ...
Nili, U.; de Wit, H.; Gulyas-Kovacs, A.; Toonen, R. F.; Soerensen, J. B.; Verhage, M.; Ashery, U.: Munc18-1 phosphorylation by protein kinase C potentiates vesicle pool replenishment in bovine chromaffin cells. Neuroscience 143 (2), pp. 487 - 500 (2006 ...
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
Reactome is pathway database which provides intuitive bioinformatics tools for the visualisation, interpretation and analysis of pathway knowledge.
The present application relates to a steam treated pelletized feed composition comprising a granule comprising a core and a coating wherein the core comprises an active compound and the coating comprises a salt.
Synaptotagmin-1, the canonical isoform of the synaptotagmin family, is a Ca(2+) sensor for fast synchronous neurotransmitter release in forebrain neurons and chromaffin cells. Even though deletion of synaptotagmin-1 abolishes fast exocytosis in chromaffin cells, it reduces overall secretion by only 20% because of the persistence of slow exocytosis. Therefore, another Ca(2+) sensor dominates release in these cells. Synaptotagmin-7 has a higher Ca(2+) affinity and slower binding kinetics than synaptotagmin-1, matching the proposed properties for the second, slower Ca(2+) sensor. Here, we examined Ca(2+)-triggered exocytosis in chromaffin cells from KO mice lacking synaptotagmin-7, and from knockin mice containing normal levels of a mutant synaptotagmin-7 whose C(2)B domain does not bind Ca(2+). In both types of mutant chromaffin cells, Ca(2+)-triggered exocytosis was decreased dramatically. Moreover, in chromaffin cells lacking both synaptotagmin-1 and -7, only a very slow release component, ...
The ACh-stimulated increase in [Ca2+]i in bovine adrenal chromaffin cells is mainly triggered by an influx of Ca2+ through the nAChR channel, VOC, and the subsequent activation of Ca2+-induced Ca2+ release, all of which contribute to CA release. These events in response to ACh are of short duration, whereas PACAP induces large and sustained increases in [Ca2+]i and CA release. The present study sought to elucidate which pathways (nAChR channel, VOC, SOC, or an unidentified channel) contribute to this peculiar Ca2+ and secretory response to PACAP.. Reports vary concerning the effect of VOC blockers on PACAP-induced rise in [Ca2+]i and CA release. For example, Przywara et al. (1996) showed that in rat cultured adrenal chromaffin cells, neither L- nor N-type VOC participates in the PACAP-induced CA release. On the other hand,Fukushima et al. (2001b) showed that nifedipine, L-type VOC antagonist, reduced PACAP-induced CA release in isolated perfused rat adrenal gland. Tanaka et al. (1996) reported ...
S. Karanth, W. H. Yu, A. Walczewska, C. Mastronardi, S. M. McCann, Ascorbic acid acts as an inhibitory transmitter in the hypothalamus to inhibit stimulated luteinizing hormone-releasing hormone release by scavenging nitric oxide, Proceedings of the National Academy of Sciences, 2000, 97, 4, ...
Chromaffin cells are neuroendocrine cells found predominantly in the medulla of the adrenal gland. They are also found in other ganglia of the sympathetic nervous system and are derived from the embryonic neural crest. Embryology They arise in ...
The properties of Ca(2+)- and voltage-dependent K+ currents and their role in defining membrane potential were studied in cultured rat chromaffin cells. Two variants of large-conductance, Ca2+ and voltage-dependent BK channels, one noninactivating and one inactivating, were largely segregated among patches. Whole-cell noninactivating and inactivating currents resulting from each of these channels were segregated among different chromaffin cells. Cell-to-cell variation in the rate and extent of whole-cell current decay was not explained by differences in cytosolic [Ca2+] regulation among cells; rather, variation was due to differences in the intrinsic properties of the underlying BK channels. About 75% of rat chromaffin cells and patches express inactivating BK current (termed BKi) while the remainder express noninactivating BK current (termed BKs). The activation time course of both currents is similar, as is the dependence of activation on [Ca2+] and membrane potential. However, deactivation of ...
The differentiation of neuronal cell progenitors depends on complex interactions between intrinsic cellular programs and environmental cues. Such interactions have recently been explored using an immortalized sympathoadrenal progenitor cell line, MAH. These studies have revealed that depolarizing conditions, in combination with exposure to FGF, can induce responsiveness to NGF. Here we report that CNTF, which utilizes an intracellular signaling pathway distinct from that of both FGF and NGF, can collaborate with FGF to promote efficiently the differentiation of MAH progenitor cells to a stage remarkably reminiscent of NGF-dependent, postmitotic sympathetic neurons. We also find that similar collaborative interactions can occur during transdifferentiation of normal cultured chromaffin cells into sympathetic neurons ...
Doxycycline synthroid. In most instances, the biopsy is taken from the actual tumor. Chlorpromazine was with- drawn and doxycycline synthroid substituted (up to a maximum of 600 mgday). Chronic morphine increases levels synthrodi types I (ACI) and VIII (ACVIII) adenylyl cyclase, PKA catalytic (C) and regulatory type II (RII) subunits, and several phosphoproteins, including CREB and tyrosine hydroxylase (TH), the rate-limiting enzyme in norepinephrine biosynthesis.
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Cleavage of the disulfide bond linking the heavy and the light chains of tetanus toxin is necessary for its inhibitory action on exocytotic release ofcatecholamines from permeabi1ized chromaffin cells [(1989) FEBS Lett. 242, 245-248; (1989) J. Neurochern., in press]. The related botulinum A toxin also consists of a heavy and a light chain linked by a disulfide bond. The actions ofboth neurotoxins on exocytosis were presently compared using streptolysin O-permeabilized bovine adrenal chromaffin cells. Botulinum A toxin inhibited Ca2 +-stimulated catecholamine release from these cells. Addition of dithiothreitollowered the effective doses to values below 5 nM. Under the same conditions, the effective doses of tetanus toxin were decreased by a factor of five. This indicates that the interchain S-S bond of botulinum A toxin must also be split before the neurotoxin can exert its effect on exocytosis. ...
Treatment of cultured bovine adrenal chromaffin cells with the catecholamine transport blocker reserpine was previously shown to increase enkephalin levels several-fold. To explore the biochemical mechanism of this effect, we examined the effect of reserpine treatment on the activities of three different peptide precursor processing enzymes: carboxypeptidase E (CPE) and the prohormone convertases (PCs) PC1/3 and PC2. Reserpine treatment increased both CPE and PC activity in extracts of cultured chromaffin cells; total protein levels were unaltered for any enzyme. Further analysis showed that the increase in CPE activity was due to an elevated Vmax, with no change in the Km for substrate hydrolysis or the levels of CPE mRNA. Reserpine activation of endogenous processing enzymes was also observed in extracts prepared from PC12 cells stably expressing PC1/3 or PC2. In vitro experiments using purified enzymes showed that catecholamines inhibited CPE, PC1/3 and PC2, with dopamine quinone the most ...
TY - JOUR. T1 - Neuropeptide Y inhibition of nicotinic receptor-mediated chromaffin cell secretion. AU - Hexum, T. D.. AU - Zheng, Jialin C. AU - Zhu, J.. PY - 1994/1/1. Y1 - 1994/1/1. N2 - Neuropeptide Y (NPY), a widely distributed peptide with varied activities, inhibits nicotinic receptor-induced [3H]norepinephrine ([3H]NE) secretion from bovine chromaffin cells. The secretion produced by membrane depolarization with high KCl concentrations or veratridine is not inhibited. Fragments of NPY, such as NPY18-36, are potent inhibitors of [3H]NE secretion, whereas [Leu31,Pro34]-NPY and peptide YY have no effect. The response to NPY18-36 is not sensitive to pertussis toxin pretreatment of chromaffin cells. NPY fragments also inhibit nicotinic receptor-induced 45Ca++ influx but not that induced by KCl or veratridine. The rank orders of potency for inhibition of [3H]NE secretion and 45Ca++ influx are the same: NPY18-36 ≥ NPY26-36 , NPY13-36. NPY and NPY(free acid) are weak inhibitors of secretion ...
The possibility of differentiating between chromaffin vesicles with different catecholamine contents was tested by studying the distribution of rabbit adrenal dopamine β-hydroxylase (EC 1.14.21) and catecholamines, and the buoyant densities of the catecholamine storage vesicles after isopycnic centrifugation of crude storage vesicle fractions in sucrose density gradients. Catecholamine storage vesicles were prepared from adrenal glands of untreated rabbits, rabbits which had received chlorisondamine chloride (10/kg intraperitoneally) to block ganglionic transmission, and rabbits which had received both chlorisondamine chloride and reserpine (1 mg kg). Adrenal glands were examined 1 day after treatmenmt with chlorisondamine and 1 and 8 days after combined treatment with chlorisondamine and reserpine. Intact storage vesicles obtained from glands of untreated animals had a specific gravity of 1.27, while the membranes obtained from vesicles lysed in distilled water had a specific gravity of 1.12. ...
Since the work of Katz, Douglas, and their collaborators almost half a century ago (Katz, 1969), a central concept in the physiology of neurosecretion is that a rise in cytosolic [Ca2+], resulting from Ca2+ influx, triggers exocytosis. More recently it has become clear that the rise in [Ca2+] occurs in a microdomain within the vicinity (i.e., at a distance of 200-300 nm in chromaffin cells) of plasmalemmal Ca2+ channels (García et al., 2006; Neher and Sakaba, 2008). This finding raises the possibility of other microdomains where a rise in focal [Ca2+] might mediate other processes, allowing Ca2+ to subserve several functions without cross talk. This possibility receives further support from the study of Ca2+ sparks in smooth muscle cells. Ca2+ sparks are focal Ca2+ transients found in striated and smooth muscle and mediated by RYRs (Cheng and Lederer, 2008). In striated muscle, they are the quanta or building blocks that make up a global increase in [Ca2+] to trigger contraction (Csernoch, ...
We have demonstrated previously that spontaneously diabetic BB-Wistar rats exhibit decreased adrenal medullary catecholamine secretion in response to splanchnic nerve terminal stimulation. We hypothesized that this abnormality is caused by changes in the sensitivity of the adrenomedullary chromaffin cells to acetylcholine (ACh). To study this hypothesis, we isolated adrenal glands from control and spontaneously diabetic BB-Wistar rats, perfused them with ACh, and measured catecholamine secretion. Adrenal catecholamine release in response to ACh was significantly decreased at 2, 8, and 16 weeks after the onset of diabetes compared with age-matched, nondiabetic control rats. Catecholamine release in response to perfusion with 20 mM K+ was the same in adrenals from diabetic and control rats. The decreased responsiveness of diabetic rat adrenals to perfusion with ACh was significantly correlated with a decrease in the release of catecholamines in response to splanchnic nerve stimulation. A similar ...
With the type of cryofixation we used (sandwich freezing), cells remain permanently surrounded by their culture or trigger medium, and impairment of viability during handling can be minimized (Pscheid et al., 1981). The propane jet applied onto the thin copper cover provides very rapid cooling, i.e., up to 40,000°C·s−1 (Knoll et al., 1982; Plattner and Knoll, 1984). The freeze-substitution medium used includes OsO4 and, thus, precludes osmotic changes during warming (Van Harreveld et al., 1965; Morel et al., 1971; Wollweber et al., 1981), and the epoxide embedding chosen involves little shrinkage (Plattner and Zingsheim, 1983). In other studies, open samples were used for freezing on cold metal surfaces or injection into cold media (Ornberg et al., 1995; Parsons et al., 1995). This requires removal of the culture/trigger medium since cryopreservation is restricted to ∼20 μm (Plattner and Bachmann, 1982). The fluid film that may remain on the sample is very thin and ionic conditions are ...
The coupling between divalent cations and exocytosis of large dense- cored vesicles (LDCV) was studied with capacitance-detection techniques in nerve terminals of the rat neurohypophysis (NHP) and bovine chromaffin cells. Ba2+ substitution for Ca2+ produced kinetically distinct responses in the two preparations. In NHP terminals, Ba2+ ions behave as weak substitutes for Ca2+. Exocytotic events occur principally during depolarizing pulses, i.e., events are "stimulus- coupled" to Ba2+ entry through voltage-gated Ca2+ channels. Stimulus- coupled exocytosis apparently requires elevated submembrane cation concentrations that dissipate rapidly on hyperpolarization-induced Ca(2+)-channel closure. Intracellular dialysis of NHP terminals with Ba2+ does not evoke exocytosis, nor does it interfere with depolarization-evoked Ca2+ influx and exocytosis. In chromaffin cells, Ba2+ ions evoke a small quantity of stimulus-coupled secretion, but the dominant response is an additional pronounced poststimulus ...
Looking for adrenomedullary hormone? Find out information about adrenomedullary hormone. secretory substance carried from one gland or organ of the body via the bloodstream to more or less specific tissues, where it exerts some influence upon... Explanation of adrenomedullary hormone
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Chromaffin cells of the adrenal medulla synthesize, store and secrete catecholamines. These cells contain numerous electron-dense secretory granules which discharge their contents into the extracellular space by exocytosis. The subplasmalemmal area of the chromaffin cell is characterized by the presence of a highly organized cytoskeletal network. F-Actin seems to be exclusively localized in this area and together with specific actin-binding proteins forms a dense viscoelastic gel; fodrin, vinculin and caldesmon, three actin cross-linking proteins, and gelsolin, an actin-severing protein, are found in this subplasmalemmal region. Since fodrin-, caldesmon- and alpha-actinin-binding sites exist on secretory granule membranes, actin filaments can also link secretory granules. Chromaffin granules can be entrapped in this subplasmalemmal lattice and thus the cytoskeleton acts as a barrier preventing exocytosis. When cells are stimulated, molecular rearrangements of the subplasmalemmal cytoskeleton ...
Marley, PD, McLeod, J, Anderson, C and Thompson, KA 1995, Nerves containing nitric oxide synthase and their possible function in the control of catecholamine secretion in the bovine adrenal medulla, Journal of the Autonomic Nervous System, vol. 54, no. 3, pp. 184-194, doi: 10.1016/0165-1838(95)00013-N. ...
Growth and development of term children born with low birth weight: effects of genetic and environmental.As mentioned above, all nutrients are important for brain development, but some appear to have a particularly large effect.The health and function of the entire body is dependent on the health of the nervous system.Golub MS, Takeuchi PT, Keen CL, Gershwin E, Hendricks AG, Lonnerdal B.I work with individuals and groups to help them meet their nutrition and health goals using.In the absence of overt microcephaly, infants with intrauterine growth retardation nevertheless have a 15% rate of mild.Dietary copper deficiency alters protein levels of rat dopamine beta-monooxygenase and tyrosine monooxygenase.. Furthermore, the abnormal behavioral function should be subserved by a brain region or process that is.Logic would dictate that sufficient nutrition and excessive treatments can be correcting what led into.Ferguson SA, Berry KJ, Hansen DK, Wall KS, White G, Antony AC.Although nutritional copper ...
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Adrenomedullary chromaffin cells have been used as an excellent experimental model to study the exocytosis and therefore the molecular mechanisms of neurotransmission. It is now clear that the proteins involved in the processes of vesicle docking, membrane fusion and neurotransmitter release are common to many cellular systems (SNARE hypothesis). Our research interest is focused in two different aspects of the molecular mechanisms of neurotransmission: Implication of molecular motors such myosin-actin in vesicle transport during neurosecretion and the determination of essential aminoacids of synaptobrevin or SNAP-25 implicated in the process of membrane fusion. Experimental approaches involve strategies using antibodies, sequence peptide design and protein overexpression that demonstrate the participation of specific protein domains in exocytosis. In addition, the role of these proteins on the secretory stages have been studied using amperometry, technique that resolves single fusion events ...
Copyright Get Revising 2017 all rights reserved. Get Revising is one of the trading names of The Student Room Group Ltd. Register Number: 04666380 (England and Wales), VAT No. 806 8067 22 Registered office: International House, Queens Road, Brighton, BN1 3XE ...
Metyrapone: An inhibitor of the enzyme STEROID 11-BETA-MONOOXYGENASE. It is used as a test of the feedback hypothalamic-pituitary mechanism in the diagnosis of CUSHING SYNDROME.
On the Convergence of Bio-, Information-, Enrivonmental-, Energy-, Space- and Nano-Technolgies: Effect of Morphine and Bupivacaine on Nicotine-Induced Catecholamine Secretion from Encapsulated Chromaffin Cells
We used the perforated-patch technique to examine the relationship between Ca2+ entry and exocytosis of large dense-cored vesicles in bovine adrenal chromaffin cells. Exocytosis evoked by single-step depolarizations was monitored by capacitance detection. Ca2+ entry was varied by changing external calcium concentration, stepping to different test potentials, depolarizing for different durations, or applying blockers of specific calcium channel subtypes. Regardless of protocol, the amount of exocytosis was strictly related to the integral of the voltage-clamped calcium current, raised to a power of approximately 1.5. Thus, despite the complexities of transient and nonuniform changes in submembrane calcium concentration produced by voltage-gated calcium entry, the calcium dependence of large dense-cored vesicle fusion under conditions of minimal stimulation is well approximated by a simple transfer function of summed calcium entry.
During exocytosis, the fusion pore expands to allow release of neurotransmitters and hormones to the extracellular space. To understand the process of synaptic transmission, it is of outstanding importance to know the properties of the fusion pore and how these properties affect the release process. Many proteins have been implicated in vesicle fusion; however, there is little evidence for proteins involved in fusion pore expansion. Myosin II has been shown to participate in the transport of vesicles and, surprisingly, in the final phases of exocytosis, affecting the kinetics of catecholamine release in adrenal chromaffin cells as measured by amperometry. Here, we have studied single vesicle exocytosis in chromaffin cells overexpressing an unphosphorylatable form (T18AS19A RLC-GFP) of myosin II that produces an inactive protein by patch amperometry. This method allows direct determination of fusion pore expansion by measuring its conductance, whereas the release of catecholamines is recorded ...
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Membranes of chromaffin granules. Isolation and partial characterization of two proteins | Biochemical JournalMembranes of chromaffin granules. Isolation and partial characterization of two proteins | Biochemical Journal

Membranes of chromaffin granules. Isolation and partial characterization of two proteins. Heide Hörtnagl, H. Winkler, J. A. L. ... Membranes of chromaffin granules. Isolation and partial characterization of two proteins. Heide Hörtnagl, H. Winkler, J. A. L. ... Membranes of chromaffin granules. Isolation and partial characterization of two proteins Message Subject (Your Name) has ... Membranes of chromaffin granules were isolated from the adrenal glands of four different species. The solubilized membrane ...
more infohttp://www.biochemj.org/content/122/3/298.1

Catecholamine Release-Inhibitory Peptide Catestatin (Chromogranin A352-372) | CirculationCatecholamine Release-Inhibitory Peptide Catestatin (Chromogranin A352-372) | Circulation

The catecholamine storage vesicle protein chromogranin A plays a role in the formation of catecholamine secretory granules and ... and regional distribution in chromaffin and nervous tissue elucidated by radioimmunoassay. J Biol Chem. 1984; 259: 3237-3247. ... 1 plays a necessary role in formation of catecholamine secretory granules.2,3 CHGA is also precursor of the catecholamine ... which acts as a nicotinic cholinergic antagonist to block transmitter release from chromaffin cells and noradrenergic nerves1,4 ...
more infohttp://circ.ahajournals.org/content/115/17/2271

Expression of the noradrenaline transporter and phenylethanolamine N-methyltransferase in normal human adrenal gland and...Expression of the noradrenaline transporter and phenylethanolamine N-methyltransferase in normal human adrenal gland and...

Immunoreactivity for CGA was colocalized with NAT within the cytoplasm of normal human chromaffin cells (n=4). This co- ... as a marker for secretory granules. Catecholamine content was measured by using high performance liquid chromatography (HPLC). ... In normal human adrenal medulla (n=5), all chromaffin cells demonstrated strong TH, PNMT and NAT immunoreactivity. NAT was co- ...
more infohttp://researchrepository.murdoch.edu.au/id/eprint/14361/

Role of phosphatidylserine and diacylglycerol in the fusion of chromaffin granules with target membranes.  - PubMed - NCBIRole of phosphatidylserine and diacylglycerol in the fusion of chromaffin granules with target membranes. - PubMed - NCBI

Role of phosphatidylserine and diacylglycerol in the fusion of chromaffin granules with target membranes.. Sánchez-Migallón MP1 ... The role of phosphatidylserine and diacylglycerol in the fusion of chromaffin granules with target membranes was investigated ... Chromaffin granule ghosts were induced to fuse with model membranes of different complexities from one resembling the inner ... Polylysine was able to induce fusion of chromaffin granule ghosts with plasma membrane vesicles in the absence of Ca2+. This ...
more infohttps://www.ncbi.nlm.nih.gov/pubmed/7944396?dopt=Abstract

Structural and Enzti1atic Studies of Dopamine-ß-Hydroxylase from Bovine Adrenal Chromaffin GranulesStructural and Enzti1atic Studies of Dopamine-ß-Hydroxylase from Bovine Adrenal Chromaffin Granules

... Albanesi, Joseph P. ... is the only enzyme of the catecholamine biosynthetic pathway located in the chromaffin granules of adrenal medulla. Within the ... granules, two populations of DßH exist: a water-soluble fraction found within the granule matrix and a membrane-bound, ...
more infohttps://dukespace.lib.duke.edu/dspace/handle/10161/7389

Identification and purification of a functional amine transporter from bovine chromaffin granules. - Semantic ScholarIdentification and purification of a functional amine transporter from bovine chromaffin granules. - Semantic Scholar

The amine transporter from bovine chromaffin granules has been purified in a functional state. Two isoforms with different pI ... Biochemistry and molecular biology of the vesicular monoamine transporter from chromaffin granules.. *J. P. Henry, David Botton ... The amine transporter from bovine chromaffin granules has been purified in a functional state. Two isoforms with different pI ... Identification and purification of a functional amine transporter from bovine chromaffin granules.. @article{ ...
more infohttps://www.semanticscholar.org/paper/Identification-and-purification-of-a-functional-Stern-Bach-Greenberg-Ofrath/138c44245f4a69587b1baadae69063751fb9d5fb

Proenkephalin-processing enzymes in chromaffin granules: model for neu by Vivian Y. Hook, Martin R. Schiller et al."Proenkephalin-processing enzymes in chromaffin granules: model for neu" by Vivian Y. Hook, Martin R. Schiller et al.

Chromaffin; Chromaffin Granules/enzymology; Cysteine Endopeptidases/metabolism; Diseases; Drugs; Endopeptidases; Enkephalins; ... Hook, V. Y., Schiller, M. R., Azaryan, A., Tezapsidis, N. (1996). Proenkephalin-processing enzymes in chromaffin granules: ...
more infohttps://digitalscholarship.unlv.edu/sls_fac_articles/185/

Kinetics of permeability changes induced by electric impulses in chromaffin granulesKinetics of permeability changes induced by electric impulses in chromaffin granules

... Lindner P, Neumann E, Rosenheck K (1977) ... Kinetics of permeability changes induced by electric impulses in chromaffin granules. Journal of Membrane Biology, 32(1), p 231 ... Lindner P, Neumann E, Rosenheck K. Kinetics of permeability changes induced by electric impulses in chromaffin granules. ... P. Lindner, E. Neumann, and K. Rosenheck, "Kinetics of permeability changes induced by electric impulses in chromaffin granules ...
more infohttps://pub.uni-bielefeld.de/publication/1774410

Synaptotagmin isoforms confer distinct activation kinetics and dynamics to chromaffin cell granules | JGPSynaptotagmin isoforms confer distinct activation kinetics and dynamics to chromaffin cell granules | JGP

Syt-1 granules fused with faster kinetics than Syt-7 granules. Overall, these findings demonstrate that chromaffin granules are ... Chromaffin granules fuse at different rates and efficacies in response to Ca2+. Both Syt-1 and Syt-7 granules fuse with high ... Syt-7 granules also show a greater tendency to fuse in clusters than Syt-1 granules, and granules harboring Syt-1 travel a ... Syt-1 granules are more mobile than Syt-7 granules. We next asked whether granules bearing either Syt-1 or Syt-7 demonstrated ...
more infohttp://jgp.rupress.org/content/149/8/763

Synaptotagmin isoforms confer distinct activation kinetics and dynamics to chromaffin cell granules | JGPSynaptotagmin isoforms confer distinct activation kinetics and dynamics to chromaffin cell granules | JGP

Syt-1 granules fused with faster kinetics than Syt-7 granules. Overall, these findings demonstrate that chromaffin granules are ... Chromaffin granules fuse at different rates and efficacies in response to Ca2+. Both Syt-1 and Syt-7 granules fuse with high ... Syt-7 granules also show a greater tendency to fuse in clusters than Syt-1 granules, and granules harboring Syt-1 travel a ... Syt-1 granules are more mobile than Syt-7 granules. We next asked whether granules bearing either Syt-1 or Syt-7 demonstrated ...
more infohttp://jgp.rupress.org/content/149/8/763?rss=1&ssource=mfr

Rapid SNARE-mediated fusion of liposomes and chromaffin granules with giant unilamellar vesicles. :: MPG.PuReRapid SNARE-mediated fusion of liposomes and chromaffin granules with giant unilamellar vesicles. :: MPG.PuRe

Rapid SNARE-mediated fusion of liposomes and chromaffin granules with giant unilamellar vesicles. ... Rapid SNARE-mediated fusion of liposomes and chromaffin granules with giant unilamellar vesicles. Witkowska, A., & Jahn, R. ( ... Our results show that liposomes and chromaffin granules fuse with GUVs containing activated SNAREs with only few milliseconds ... chromaffin granules with giant unilamellar vesicles. Biophysical Journal, 113(6), 1251-1259. doi:10.1016/j.bpj.2017.03.010. ...
more infohttps://pure.mpg.de/pubman/faces/ViewItemFullPage.jsp?itemId=item_2431935

Chromogranin A-processing proteinases in purified chromaffin granules: contaminants or endogenous enzymes?<...Chromogranin A-processing proteinases in purified chromaffin granules: contaminants or endogenous enzymes?<...

A trypsin-like peptidase, most active at pH 8.2, was enriched in a membrane wash of chromaffin granules. Subcellular ... A trypsin-like peptidase, most active at pH 8.2, was enriched in a membrane wash of chromaffin granules. Subcellular ... A trypsin-like peptidase, most active at pH 8.2, was enriched in a membrane wash of chromaffin granules. Subcellular ... A trypsin-like peptidase, most active at pH 8.2, was enriched in a membrane wash of chromaffin granules. Subcellular ...
more infohttps://einstein.pure.elsevier.com/en/publications/chromogranin-a-processing-proteinases-in-purified-chromaffin-gran-2

The Association of Dynamin with Synaptophysin Regulates Quantal Size and Duration of Exocytotic Events in Chromaffin Cells |...The Association of Dynamin with Synaptophysin Regulates Quantal Size and Duration of Exocytotic Events in Chromaffin Cells |...

... other proteins present in the granules, such as syntaxin, contributing to the association of dynamin to chromaffin granules ( ... confirming its association with chromaffin granules. A similar distribution of dynamin in bovine chromaffin cells was ... Chromaffin granules were purified from the 20,000 × g pellet using 1.6 m sucrose gradients (Smith and Winkler, 1967). The ... Indeed, our results show that synaptophysin and dynamin are present in chromaffin granules (Fig. 2A,B), and that both anti-Syn ...
more infohttp://www.jneurosci.org/content/30/32/10683

Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells<...Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells<...

After exocytosis, chromaffin granules release essentially all their catecholamines in small fractions of a second, but it is ... N2 - After exocytosis, chromaffin granules release essentially all their catecholamines in small fractions of a second, but it ... AB - After exocytosis, chromaffin granules release essentially all their catecholamines in small fractions of a second, but it ... abstract = "After exocytosis, chromaffin granules release essentially all their catecholamines in small fractions of a second, ...
more infohttps://ohsu.pure.elsevier.com/en/publications/recapture-after-exocytosis-causes-differential-retention-of-prote-2

Plus itPlus it

Catecholamine release from chromaffin granules is unchanged in TRP mutants. A, Schematic drawing of the experimental setup. B, ... A-C, Wild-type chromaffin cells and dko cells expressing either SybII or the WWAA mutant exhibit SybII-positive granules ( ... We found that deletion or substitution of these Trp residues reduces the pool size of primed chromaffin granules, but leaves ... Membrane-proximal Trp residues are not essential for rapid transmitter discharge from chromaffin granules. To determine the ...
more infohttp://www.jneurosci.org/content/32/45/15983

Comparison of cysteine string protein (Csp) and mutant alpha-SNAP overexpression reveals a role for csp in late steps of...Comparison of cysteine string protein (Csp) and mutant alpha-SNAP overexpression reveals a role for csp in late steps of...

... of overexpression of a dominant negative alpha-SNAP mutant or Csp on exocytosis of dense-core granules in single chromaffin ... Exocytosis of trans-Golgi network (TGN)-derived dense-core granules was substantially inhibited by expression of alpha-SNAP( ... Delay in vesicle fusion revealed by electrochemical monitoring of single secretory events in adrenal chromaffin cells. *Robert ... is essential for exocytosis in chromaffin cells.. *María Teresa Criado, A. Luisa Gil, Salvador Viniegra, Luis M. Gutíerrez ...
more infohttps://www.semanticscholar.org/paper/Comparison-of-cysteine-string-protein-%28Csp%29-and-a-Graham-Burgoyne/36ceb82e6f9ccf2c591b4d4fd2e1c3e88c3f7c8a

ATPase - WikipediaATPase - Wikipedia

Njus, D.; Knoth, J.; Zallakian, M. (1981). "Proton-linked transport in chromaffin granules". Curr. Top. Bioenerg. 11: 107-147. ...
more infohttps://en.wikipedia.org/wiki/ATPase

Protein kinase C enhances exocytosis from chromaffin cells by increasing the size of the readily releasable pool of secretory...Protein kinase C enhances exocytosis from chromaffin cells by increasing the size of the readily releasable pool of secretory...

... kinase C enhances exocytosis from chromaffin cells by increasing the size of the readily releasable pool of secretory granules. ... Three different protocols were used to show that PMA increases the size of the readily releasable pool of secretory granules. ...
more infohttps://www.ncbi.nlm.nih.gov/pubmed/8663997?dopt=Abstract

Recombinant Human SYT2 protein (ab165289) | AbcamRecombinant Human SYT2 protein (ab165289) | Abcam

Cytoplasmic vesicle , secretory vesicle , chromaffin granule membrane. Synaptic vesicles and chromaffin granules. ...
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Anti-Synaptotagmin 1 antibody (ab106621) | AbcamAnti-Synaptotagmin 1 antibody (ab106621) | Abcam

Cytoplasmic vesicle , secretory vesicle , chromaffin granule membrane. Cytoplasm. Synaptic vesicles and chromaffin granules. ...
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ATPase V - Wikipedia, a enciclopedia libreATPase V - Wikipedia, a enciclopedia libre

ATPase of chromaffin granules. Kinetics, regulation, and stoichiometry". J. Biol. Chem. 257 (18): 10701-7. PMID 6213624.. ...
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First Pass Miss Exam 3 Flashcards by Matthew Miller | BrainscapeFirst Pass Miss Exam 3 Flashcards by Matthew Miller | Brainscape

Dopamine -> norepinephrine in chromaffin granule. NE -> E in cytoplasm.. E and NE stored in chromaffin granules at 3:1 ratio ... Increases E/NE production, but not release, by activating two key hydroxylase enzymes in chromaffin cells. ...
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SYT2 Gene - GeneCards | SYT2 Protein | SYT2 AntibodySYT2 Gene - GeneCards | SYT2 Protein | SYT2 Antibody

synaptotagmin 2,multifunctional intrinsic membrane protein of synaptic vesicles and chromaffin granules,involved in exocytosis ... Cytoplasmic vesicle, secretory vesicle, chromaffin granule membrane; Single-pass membrane protein. Note=Synaptic vesicles and ...
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Organelle Analysis - Current ProtocolsOrganelle Analysis - Current Protocols

Isolation of Chromaffin Granules. Carl E. Creutz. Published online: September 2010. Isolation of Cytotoxic T Cell and NK ... Isolation of Neuromelanin Granules. Florian Tribl. Published online: December 2008. Complex I Assay in Mitochondrial ... Isolation of Platelet Granules. Juliane Nießen, Gabriele Jedlitschky, Andreas Greinacher, Heyo K. Kroemer. Published online: ... Granules and Purification of Their Effector Proteins. Jerome Thiery, Michael Walch, Danielle K. Jensen, Denis Martinvalet, Judy ...
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what are the four cytoplasmic granules. chromaffin granules, endocrine granules, argentaffin granules, paneth granules. ... give facts about endogenous nonhematogenous cytoplasmic granules. can be demonstrated via argyrophil and/or argentaffin ... hematogenous (blood derived), non-hematogenous (melanin, lipofuchsin, chromaffin), endogenous minerals (Ca, Cu, uric acid and ...
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  • Chromogranin A-processing proteinases in purified chromaffin granules: contaminants or endogenous enzymes? (elsevier.com)
  • Upon depolarization of cells expressing fluorescent Syt isoforms using elevated K + , we find that Syt-7 granules fuse with faster kinetics than Syt-1 granules, irrespective of stimulation strength. (rupress.org)
  • Syt-7 granules also show a greater tendency to fuse in clusters than Syt-1 granules, and granules harboring Syt-1 travel a greater distance before fusion than those with Syt-7, suggesting that there is spatial and fusion-site heterogeneity among the two granule populations. (rupress.org)
  • Upon introduction of Ca 2+ into permeabilized cells, Syt-7 granules fuse with fast kinetics and high efficacy, even at low Ca 2+ levels (e.g., when cells are weakly stimulated). (rupress.org)
  • Our results show that liposomes and chromaffin granules fuse with GUVs containing activated SNAREs with only few milliseconds delay between docking and fusion. (mpg.de)
  • Taking advantage of the dependence on pH of the fluorescence of green fluorescent protein, we used rhythmic external acidification to determine whether and when granules re-sealed. (elsevier.com)
  • Proenkephalin-processing enzymes in chromaffin granules: model for neu" by Vivian Y. Hook, Martin R. Schiller et al. (unlv.edu)
  • Proenkephalin-processing enzymes in chromaffin granules: model for neuropeptide biosynthesis. (unlv.edu)
  • Two enzymes with low activity (a Ca 2+ activated proteinase and a trypsin-like enzyme) are, apparently, true constituents of chromaffin granules. (elsevier.com)
  • Within the granules, two populations of DßH exist: a water-soluble fraction found within the granule matrix and a membrane-bound, amphiphilic fraction embedded in the surrounding bilayer. (duke.edu)
  • This enzyme which was inhibited by p-chloromercuriphenylsulfonic acid was localized in chromaffin granules. (elsevier.com)
  • The distribution in the degree of filling of granules that had been partially depleted by an electric field pulse indicated that the population could be considered homogeneous with respect to release. (uni-bielefeld.de)
  • Here we compare the exocytic release of fluorescently labelled neuropeptide Y (NPY) and tissue plasminogen activator from single granules. (elsevier.com)
  • Conversely, Syt-1 granules require a comparatively larger increase in intracellular Ca 2+ for activation. (rupress.org)