GABAergic excitatory synapses and electrical coupling sustain prolonged discharges in the prey capture neural network of Clione limacina.
Afterdischarges represent a prominent characteristic of the neural network that controls prey capture reactions in the carnivorous mollusc Clione limacina. Their main functional implication is transformation of a brief sensory input from a prey into a lasting prey capture response. The present study, which focuses on the neuronal mechanisms of afterdischarges, demonstrates that a single pair of interneurons [cerebral A interneuron (Cr-Aint)] is responsible for afterdischarge generation in the network. Cr-Aint neurons are electrically coupled to all other neurons in the network and produce slow excitatory synaptic inputs to them. This excitatory transmission is found to be GABAergic, which is demonstrated by the use of GABA antagonists, uptake inhibitors, and double-labeling experiments showing that Cr-Aint neurons are GABA-immunoreactive. The Cr-Aint neurons organize three different pathways in the prey capture network, which provide positive feedback necessary for sustaining prolonged spike activity. The first pathway includes electrical coupling and slow chemical transmission from the Cr-Aint neurons to all other neurons in the network. The second feedback is based on excitatory reciprocal connections between contralateral interneurons. Recurrent excitation via the contralateral cell can sustain prolonged interneuron firing, which then drives the activity of all other cells in the network. The third positive feedback is represented by prominent afterdepolarizing potentials after individual spikes in the Cr-Aint neurons. Afterdepolarizations apparently represent recurrent GABAergic excitatory inputs. It is suggested here that these afterdepolarizing potentials are produced by GABAergic excitatory autapses. (+info)
Determination of the anomeric configurations of Corbicula ceramide di- and trihexoside by chromium trioxide oxidation.
The anomeric configurations of Corbicula ceramide dihexoside and ceramide trihexoside were determined by chromium trioxide oxidation and the structures of these lipids were shown to be Man-beta(1 leads to 4)-Glc-beta(1 leads to 1)-ceramide and Man-alpha(1 leads to 4)-Man-beta(1 leads to 4)-Glc-beta(1 leads to 1)-ceramide. These results are compatible with those obtained by enzymic hydrolysis reported previously. (+info)
Central pattern generator for escape swimming in the notaspid sea slug Pleurobranchaea californica.
Escape swimming in the notaspid opisthobranch Pleurobranchaea is an episode of alternating dorsal and ventral body flexions that overrides all other behaviors. We have explored the structure of the central pattern generator (CPG) in the cerebropleural ganglion as part of a study of neural network interactions underlying decision making in normal behavior. The CPG comprises at least eight bilaterally paired interneurons, each of which contributes and is phase-locked to the swim rhythm. Dorsal flexion is mediated by hemiganglion ensembles of four serotonin-immunoreactive neurons, the As1, As2, As3, and As4, and an electrically coupled pair, the A1 and A10 cells. When stimulated, A10 commands fictive swimming in the isolated CNS and actual swimming behavior in whole animals. As1-4 provide prolonged, neuromodulatory excitation enhancing dorsal flexion bursts and swim cycle number. Ventral flexion is mediated by the A3 cell and a ventral swim interneuron, IVS, the soma of which is yet unlocated. Initiation of a swim episode begins with persistent firing in A10, followed by recruitment of As1-4 and A1 into dorsal flexion. Recurrent excitation within the As1-4 ensemble and with A1/A10 may reinforce coactivity. Synchrony among swim interneuron partners and bilateral coordination is promoted by electrical coupling among the A1/A10 and As4 pairs, and among unilateral As2-4, and reciprocal chemical excitation between contralateral As1-4 groups. The switch from dorsal to ventral flexion coincides with delayed recruitment of A3, which is coupled electrically to A1, and with recurrent inhibition from A3/IVS to A1/A10. The alternating phase relation may be reinforced by reciprocal inhibition between As1-4 and IVS. Pleurobranchaea's swim resembles that of the nudibranch Tritonia; we find that the CPGs are similar in many details, suggesting that the behavior and network are primitive characters derived from a common pleurobranchid ancestor. (+info)
A novel small protein associated with a conjugated trienoic chromophore from membranes of scallop adductor muscle: phosphorylation by protein kinase A.
Membranes enriched in sarcolemma from the cross-striated adductor muscle of the deep sea scallop have been found to contain a previously undescribed small protein of 6-8 kDa that can be released by treatment with organic solvent mixtures. This proteolipid co-purified with a non-amino acid chromophore containing a conjugated trienoic moiety. Although common in plants and algae, such a stable conjugated trienoic group is unusual for an animal cell. The N-terminal amino acid sequence of the protein was XEFQHGLFGXF/ADNIGLQ, which most strongly resembles sequences in the triacyl glycerol lipase precursor and the product of the human breast cancer susceptibility gene BRCA 1, but does not show similarity to previously described proteolipids. The protein was found to be one of the major substrates in its parent membrane for the catalytic subunit of protein kinase A, which may imply a regulatory function for this molecule. (+info)
Osmotic adjustment in an estuarine population of Urosalpinx cinerea (Say, 1822) (Muricidae, Gastropoda).
Individuals from a subtidal, estuarine population of the common oyster drill, Urosalpinx cinerea (Say, 1822), were brought into the laboratory and tested for osmotic adjustment to changing salinity. Tissue variables monitored at seven experimental salinities ranging from 10 to 40% were tissue fluid osmolality, chloride, sodium, potassium, free amino acids (FAA), ninhydrin-positive substances (NPS) and water content. The results of this study demonstrate that the test animals did not exhibit anisosmotic regulation at any of the experimental salinities. However, the data do suggest a high degree of hyper-ionic regulation of potassium at all experimental salinities and a hyporegulation of sodium between the 25 and 40% salinities. Taurine, aspartic acid, alanine and glycine were the four FAA present in relatively consistent high amounts. These four amino acids comprised from 59.6 to 75.7% of the total FAA pools. It is postulated that the population does not maintain its euryhaline survival status through an osmoregulatory mechanism. Rather, the population has probably adapted physiologically to withstand dilution of its body fluids during spring conditions of low salinities. (+info)
Fluorescence measurements detect changes in scallop myosin regulatory domain.
Ca2+-induced conformational changes of scallop myosin regulatory domain (RD) were studied using intrinsic fluorescence. Both the intensity and anisotropy of tryptophan fluorescence decreased significantly upon removal of Ca2+. By making a mutant RD we found that the Ca2+-induced fluorescence change is due mainly to Trp21 of the essential light chain which is located at the unusual Ca2+-binding EF-hand motif of the first domain. This result suggests that Trp21 is in a less hydrophobic and more flexible environment in the Ca2+-free state, supporting a model for regulation based on the 2 A resolution structure of scallop RD with bound Ca2+ [Houdusse A. and Cohen C. (1996) Structure 4, 21-32]. Binding of the fluorescent probe, 8-anilinonaphthalene-1-sulphonate (ANS) to the RD senses the dissociation of the regulatory light chain (RLC) in the presence of EDTA, by energy transfer from a tryptophan cluster (Trp818, 824, 826, 827) on the heavy chain (HC). We identified a hydrophobic pentapeptide (Leu836-Ala840) at the head-rod junction which is required for the effective energy transfer and conceivably is part of the ANS-binding site. Extension of the HC component of RD towards the rod region results in a larger ANS response, presumably indicating changes in HC-RLC interactions, which might be crucial for the regulatory function of scallop myosin. (+info)
The occurrence of two types of collagen proalpha-chain in the abalone Haliotis discus muscle.
Acid-soluble collagens were prepared from connective tissues in the abalone Haliotis discus foot and adductor muscles with limited proteolysis using pepsin. Collagen preparation solubilized with 1% pepsin contained two types of alpha-chains which were different in their N-terminal amino acid sequences. Accordingly, two types of full-length cDNAs coding for collagen proalpha-chains were isolated from the foot muscle of the same animal and these proteins were named Hdcols (Haliotis discus collagens) 1alpha and 2alpha. The two N-terminal amino acid sequences of the abalone pepsin-solubilized collagen preparation corresponded to either of the two sequences deduced from the cDNA clones. In addition, several tryptic peptides prepared from the pepsin-solubilized collagen and fractionated by HPLC showed N-terminal amino acid sequences identical to those deduced from the two cDNA clones. Hdcols 1alpha and 2alpha consisted of 1378 and 1439 amino acids, respectively, showing the primary structure typical to those of fibril-forming collagens. The N-terminal propeptides of the two collagen proalpha-chains contained cysteine-rich globular domains. It is of note that Hdcol 1alpha completely lacked a short Gly-X-Y triplet repeat sequence in its propeptide. An unusual structure such as this has never before been reported for any fibril-forming collagen. The main triple-helical domains for both chains consisted of 1014 amino acids, where a supposed glycine residue in the triplet at the 598th position from the N-terminus was replaced by alanine in Hdcol 1alpha and by serine in Hdcol 2alpha. Both proalpha-chains of abalone collagens contained six cysteine residues in the carboxyl-terminal propeptide, lacking two cysteine residues usually found in vertebrate collagens. Northern blot analysis demonstrated that the mRNA levels of Hdcols 1alpha and 2alpha in various tissues including muscles were similar to each other. (+info)
Continuous in vitro propagation and differentiation of cultures of the intramolluscan stages of the human parasite Schistosoma mansoni.
The metazoan parasitic blood flukes, Schistosoma spp., infect over 200 million people worldwide and cause extensive human morbidity and mortality. Research strategies for development of anti-schistosomal agents are impeded by the organism's complex molluscan-mammalian life cycle, which limits experimental approaches and availability of material. We derived long-term continuously proliferative cultures of Schistosoma mansoni sporocysts capable of generating cercariae in vitro. Cultured organisms retained the ability to parasitize the host, and they exhibited developmental regulation of candidate stage-specific genes in the host-free culture system. Evidence for expression of a reverse transcriptase also was found in the cultured organisms, pointing to this activity as a possible mechanistic contributor to the dynamic relationship between the parasite and its hosts. Continuous in vitro propagation of the asexual sporocyst stage allows isolation of clonally derived parasite populations and provides a means to study schistosomal molecular genetics, metabolism, and evasion of host defenses. (+info)