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, ...
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, ...
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
In bovine adrenal chromaffin cells, prostaglandin E2 (PGE2) stimulates the formation of inositol phosphates and Ca2+ mobilization through its specific receptor [Yokohama, Tanaka, Ito, Negishi, Hayashi & Hayaishi (1988) J. Biol. Chem. 263, 1119-1122]. Here we show that PGE2-induced phosphoinositide metabolism was blocked by pretreatment with 12-O-tetradecanoylphorbol 13-acetate (TPA). Using intact cells, we also examined the inhibitory effect of TPA on the individual steps of the activation process of phosphoinositide metabolism. The inhibition was observed within 1 min and complete by 10 min after addition of 1 microM-TPA, and half-maximal inhibition by TPA occurred at 20 nM. TPA prevented Ca2+ mobilization induced by PGE2, but not by the Ca2+ ionophore ionomycin. The inactive phorbol ester 4 alpha-phorbol 12,13-didecanoate did not inhibit the formation of inositol phosphates and Ca2+ mobilization by PGE2. TPA treatment affected neither the high-affinity binding of [3H]PGE2 to intact cells and ...
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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 ...
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
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 ...
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 ...
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
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 ...
Adrenal chromaffin cells are excitable neuroendocrine cells that have been widely used as a simple model of neurosecretion. In vivo, acetylcholine released from preganglionic neurons binds to nicotinic receptors, which are Na+ ionophores, causing Na+ influx that depolarizes the plasma membrane. Depolarization in turn causes voltage-gated calcium channels (VGCCs) to open, leading to an influx of Ca2+ that activates the fusion of secretory granules with the plasma membrane, resulting in catecholamine release that occurs within milliseconds. This Ca2+-dependent secretory process is referred to as exocytosis. Previous investigations exploring the potential for nanosecond electric pulses (NEPs) to serve as a novel bioelectric stimulus of neurosecretion in chromaffin cells have shown that in chromaffin cells exposed to 5 ns, 5 MV/m electric pulses, catecholamine release is stimulated in a manner that relies on Ca2+ influx via VGCCs. The goal of the present study was to further understand this novel ...
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 ...
TY - JOUR. T1 - The effect of ACTH, renin, angiotensin II, and various precursors on biosynthesis of aldosterone by adrenal slices.. AU - Kaplan, Norman M. AU - BARTER, F. C.. PY - 1962/4. Y1 - 1962/4. UR - http://www.scopus.com/inward/record.url?scp=0001053852&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=0001053852&partnerID=8YFLogxK. U2 - 10.1172/JCI104530. DO - 10.1172/JCI104530. M3 - Article. C2 - 14453776. AN - SCOPUS:0001053852. VL - 41. SP - 715. EP - 724. JO - Journal of Clinical Investigation. JF - Journal of Clinical Investigation. SN - 0021-9738. ER - ...
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 ...
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. We coined the term "Molecular cytoarchitecture of exocytosis" to define the interaction between SNARE proteins, calcium channel and lately nicotinic receptors (integrating Dr. Criado main line) and the cohesive F-actin cortical network in order to improve secretory efficiency ...
Video articles in JoVE about adrenal glands include A Novel Method: Super-selective Adrenal Venous Sampling, Monitoring the Effect of Osmotic Stress on Secretory Vesicles and Exocytosis, Methods for Cell-attached Capacitance Measurements in Mouse Adrenal Chromaffin Cell, Mouse Adipose Tissue Collection and Processing for RNA Analysis, Imaging Plasma Membrane Deformations With pTIRFM, An Alternant Method to the Traditional NASA Hindlimb Unloading Model in Mice, Pre-Conditioning the Airways of Mice with Bleomycin Increases the Efficiency of Orthotopic Lung Cancer Cell Engraftment, 5/6th Nephrectomy in Combination with High Salt Diet and Nitric Oxide Synthase Inhibition to Induce Chronic Kidney Disease in the Lewis Rat, Isolation and Transplantation of Different Aged Murine Thymic Grafts., In Vivo Model for Testing Effect of Hypoxia on Tumor Metastasis, Receptor Autoradiography Protocol for the Localized Visualization of Angiotensin II Receptors, Murine Prostate Micro-dissection
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 ...
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.
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 ...
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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 ...
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. ...
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, ...
MUNC18-1 (also known as STXBP1) is an essential protein for docking and fusion of secretory vesicles. Mouse chromaffin cells (MCCs) lacking MUNC18-1 show impaired secretory vesicle docking, but also mistargeting of SNARE protein syntaxin1 and an abnormally dense submembrane F-actin network. Here, we tested the contribution of both these phenomena to docking and secretion defects in MUNC18-1-deficient MCCs. We show that an abnormal F-actin network and syntaxin1 targeting defects are not observed in Snap25- or Syt1-knockout (KO) MCCs, which are also secretion deficient. We identified a MUNC18-1 mutant (V263T in β-sheet 10) that fully restores syntaxin1 targeting but not F-actin abnormalities in Munc18-1-KO cells. MUNC18-2 and -3 (also known as STXBP2 and STXBP3, respectively), which lack the hydrophobic residue at position 263, also did not restore a normal F-actin network in Munc18-1-KO cells. However, these proteins did restore the normal F-actin network when a hydrophobic residue was ...
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,
Area of interest: Mechanisms of stress transduction at the sympatho-adrenal synapse; optical studies of hormone trafficking and secretion in the adrenomedullary chromaffin cell.
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 ...
Ahnert-Hilger, G.; Wegenhorst, U.; Stecher, B.; Spicher, K.; Rosenthal, W.; Gratzl, Manfred (1992): Exocytosis from permeabilized bovine adrenal chromaffin cells is differently modulated by guanosine 5-[gamma-thio]triphosphate and guanosine 5-[beta gamma-imido]triphosphate. Evidence for the involvement of various guanine nucleotide-binding proteins. In: Biochemical Journal, Vol. 284: pp. 321-326 [PDF, 3MB] ...
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