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R P Croll

Publications and source records attributed to R P Croll.

At least 19 recordsLinked to original sources

Catecholamine-containing cells in the central nervous system and periphery of Aplysia californica.

Previous studies have suggested the presence of numerous catecholamine-containing cells in both the central ganglia and peripheral tissues of Aplysia, but they often offered conflicting or incomplete accounts of numbers, locations, and morphologies. The current study combines aldehyde-induced histofluorescence and tyrosine hydroxylase-like immunoreactivity together with confocal microscopy to provide details of these cells. Approximately 35-50 neurones in the cerebral ganglia, 4-8 neurones in the pedal ganglia, 5 neurones in the buccal ganglia, and numerous small fibres in various nerve trunks exhibited both immunoreactivity and aldehyde-induced fluorescence. Approximately 20 cells in the pedal ganglia and 4 cells in the buccal ganglia exhibited only immunoreactivity whereas 15-20 neurons in the cerebral ganglia exhibited only aldehyde-induced fluorescence. No somata in the pleural or abdominal ganglia exhibited aldehyde-induced fluorescence or immunoreactivity. Both aldehyde-induced histofluorescence and immunoreactivity also labelled what appeared to be two classes of catecholamine-containing cells in the gill, siphon, oesophagus, rhinophore, tentacle, and reproductive organs. The more numerous, but smaller cells had subepithelial somata and processes penetrating the overlying body wall, thus suggesting a sensory function. Another class of neurones had larger somata, often located more deeply within the tissue, and occasionally appeared to be multipolar. Processes from these various peripheral cells appeared to comprise the major component of afferent fibres and to form an extensive peripheral plexus, often associated with various muscles. The morphologies of the peripheral cells thus suggest involvement in both local and centrally mediated reflexes and responses, but additional studies must test such hypothesised functions and determine the sensory modalities that the cells mediate.

Animals↗

Histochemical survey of transmitters in the central ganglia of the gastropod mollusc Phestilla sibogae.

The aeolid nudibranch Phestilla sibogae is well studied in terms of its larval nervous system and neuronal involvement in metamorphosis. Central neurones in the adult have also been identified anatomically and electrophysiologically. We describe the neurotransmitter contents of these neurones and provide details of neuritic projections and developmental changes during growth (3 to 18 mm body length). Central ganglia from specimens of all sizes contained 100-115 serotonin-immunoreactive neurones, some of which appeared to be homologues of cells identified in other gastropods. Tyrosine hydroxylase immunoreactivity and aldehyde-induced fluorescence marked a common set of 28-30 catecholaminergic neurones located anteriorly in the cerebropleural ganglia and laterally in the pedal ganglia. Ganglionic neuropile and nerve trunks also contained many catecholaminergic fibres. About 65-100 intensely labelled FMRFamide-immunoreactive neurones were located symmetrically throughout the central ganglia, although one population was located only in the right pedal ganglion. Another 40-45 FMRFamide-immunoreactive neurones were weakly or variably stained. Central ganglia also contained 27-29 intensely labelled pedalpeptide-immunoreactive neurones, including those that were apparently homologues of cells previously described in Tritonia diomedea, and 16-19 weakly labelled pedal-peptide-immunoreactive neurones, including giant cerebropleural neurones coexhibiting FMRFamide immunoreactivity. Little cell addition involving any transmitter phenotype occurred as animals grew in body length, body growth being accommodated by growth in the size of individual cells, consistent with an approximate doubling in the size of the ganglia themselves.

Aldehydes↗

Insights into early molluscan neuronal development through studies of transmitter phenotypes in embryonic pond snails.

Pond snails have long been the subject of intense scrutiny by researchers interested in general principles of development and also cellular and molecular neurobiology. Recent work has exploited both these fields of study by examining the ontogeny of the nervous system in these animals. Much of this work has focussed upon the development of specific transmitter phenotypes to provide vignettes of neuronal subpopulations that can be traced from early embryonic life through to adulthood. While such studies have generally confirmed previous explanations of gangliogenesis in gastropods, they have also indicated the presence of several neurons that appear earlier and in positions inconsistent with classical views of gastropods neurogenesis. The earliest of these cells contain FMRFamide-related peptides and have anteriorly projections that mark the future locations of ganglia and interconnecting pathways that will comprise the postembryonic central nervous system. These posterior, peptidergic cells, as well as certain, apical, monoaminergic neurons, disappear and apparently die near the end of embryonic life. Finally, populations of what appear to be peripheral sensory neurons begin to express catecholamines by around midway through embryonic life. Like several of the neurons expressing a variety of transmitters in the developing central ganglia, the catecholaminergic peripheral cells persist into postembryonic life. Transmitter phenotypes, cell shapes and locations, and neuritic morphologies all suggest that many of the neurons observed in early embryonic pond snails have recognizable homologues across the molluscs. Such observations have profoundly altered our views of neurogenesis in gastropods over the last few years. They also suggest the promise for pond snails as fruitful models for studying the roles and mechanisms for pioneering fibres, cues triggering apoptosis, and contrasting origins and mechanisms employed for generating central vs. peripheral neurons within a single organism.

Animals↗

Catechol concentrations in the hemolymph of the scallop, Placopecten magellanicus.

Catecholamines have previously been detected in numerous tissues and are thought to control a wide variety of physiological functions in bivalve molluscs. In the present study, alumina extraction and high-performance liquid chromatography reveal the presence of significant concentrations of 3,4-dihydroxyphenylalanine (DOPA), dopamine, and 3,4-dihydroxyphenylacetic acid (DOPAC) in the hemolymph of the sea scallop, Placopecten magellanicus. The concentration of dopamine in the hemolymph averaged 223.8 ng/ml, (+/-48.4, SEM), equivalent to 10(-7) to 10(-6) M. Neither epinephrine nor norepinephrine was reliably detected in significant quantities. Previous studies have demonstrated physiological responses to dopamine with thresholds of 10(-9) to 10(-6) M, thus suggesting that this catecholamine may have an endocrine function. Furthermore, monitoring hemolymph concentrations of catechols might provide a sensitive measure of the physiological status of bivalves. For example, drugs known to affect catechol concentrations in other tissues also effect hemolymph levels. Administration of monoamine oxidase inhibitors such as pargyline, deprenyl, and clorgyline at 10(-4) M for 1 day of incubation followed by a 2-day wash resulted in decreased hemolymph concentrations of DOPAC and increased concentrations of its precursors, DOPA and dopamine. Incubation in 10(-4) M 3,5-dinitrocatechol, a catecholamine-O-methyl transferase blocker, for 1 day followed by a 2-day wash significantly increased the concentration of dopamine and DOPAC in the hemolymph. Scallops incubated in 10(-5) M alpha-methyl-p-tyrosine, a blocker of tyrosine hydroxylase, for 1 day followed by a 3-day wash in artificial seawater had significantly reduced concentrations of DOPA, dopamine, and DOPAC in the hemolymph. In addition to responding to pharmacological agents, dopamine levels also decreased significantly following thermal induction of spawning, thus suggesting that hemolymph concentrations of catechols might provide indices of reproductive activity and/or stress.

3,4-Dihydroxyphenylacetic Acid↗

Catecholamines modulate metamorphosis in the opisthobranch gastropod Phestilla sibogae.

Larvae of the nudibranch Phestilla sibogae are induced to metamorphose by a factor from their adult prey, the coral Porites compressa. Levels of endogenous catecholamines increase 6 to 9 days after fertilization, when larvae become competent for metamorphosis. Six- to nine-day larvae, treated with the catecholamine precursor L-DOPA (0.01 mM for 0.5 h), were assayed for metamorphosis in response to coral inducer and for catecholamine content by high-performance liquid chromatography. L-DOPA treatment caused 20- to 50-fold increases in dopamine, with proportionally greater increases in younger larvae, so that L-DOPA-treated larvae of all ages contained similar levels of dopamine. A much smaller (about twofold) increase in norepinephrine occurred in all larvae. The treatment significantly potentiated the frequency of metamorphosis of 7- to 9-d larvae at low concentrations of inducer. In addition, L-DOPA treatment at 9 d increased aldehyde-induced fluorescence in cells that were also labeled in the controls, and revealed additional cells. However, all labeled cells were consistent with the locations of cells showing tyrosine-hydroxylase-like immunoreactivity. Catecholamines are likely to modulate metamorphosis in P. sibogae, but rising levels of catecholamines around the time of competence are insufficient alone to account for sensitivity to inducer in competent larvae.

Animals↗

Development of embryonic cells containing serotonin, catecholamines, and FMRFamide-related peptides in Aplysia californica.

This study demonstrates the presence of a relatively extensive but previously unrecognized nervous system in embryonic stages of the opisthobranch mollusc Aplysia californica. During the trochophore stage, two pairs of cells were observed to be reactive to antibodies raised against the neuropeptides FMRFamide and EFLRIamide. These cells were located in the posterior region of the embryo, and their anterior projections terminated under the apical tuft. As the embryos developed into veliger stages, serotonin-like immunoreactive (LIR) cells appeared in the apical organ and were later observed to innervate the velum. Also, aldehyde-induced fluorescence indicative of catecholamines was present in cells in the foot, oral, and possibly apical regions during late embryonic veliger stages. Just before the embryo hatches as a free-swimming veliger, additional FMRFamide-LIR and catecholamine-containing cells appeared in regions that correspond to the ganglia of what will become the adult central nervous system (CNS). Neurons and connectives that will contribute to the adult CNS appear to develop along the pathways that are pioneered by the earliest posterior FMRFamide-LIR cells. These observations are consistent with the hypothesis that, besides their presumed roles in the control of embryonic behaviors, some elements may also guide the development of the CNS. Embryonic nervous systems that develop prior to and outside of the adult CNS have also been reported in pulmonate and prosobranch species of molluscs. Therefore, the demonstration of early developing neurons and their transmitter phenotypes in A. californica presents new opportunities for a better understanding of the ontogeny and phylogeny of both behavioral and neuronal function in this important model species.

Amino Acid Sequence↗

Regional distribution and extrinsic innervation of intrinsic cardiac neurons in the guinea pig.

Mammalian intrinsic cardiac neurons subserve different functions in different cardiac regions, but the regional anatomical organisation of the intracardiac nervous system is not well understood. We investigated the quantitative and qualitative distribution of cholinergic and adrenergic elements, and the intracardiac pathways of extrinsic cardiac nerves, in whole-mount preparations of guinea pig atria. Protein gene product 9.5 immunoreactivity (PGP 9.5-IR) marked intracardiac neuronal elements; immunoreactions for choline acetyltransferase (ChAT-IR) and tyrosine hydroxylase (TH-IR) distinguished cholinergic and adrenergic components, respectively. Catecholamine-containing components were identified by aldehyde-induced fluorescence histochemistry. Mean total number of atrial neurons was 1510+/-251 (SE); 85% of these occurred in ganglia of < or = 20 neurons. All neuronal somata expressing PGP 9.5-IR also expressed ChAT-IR, suggesting that these neurons were cholinergic. Right (RA) and left (LA) atria had statistically similar neuronal densities (6.4+/-1.2 and 2.4+/-0.7 neurons/mm2, respectively; analysis of variance, P< or =0.05). Neurons in RA were concentrated intercavally; LA neurons were concentrated near pulmonary vein ostia. Greatest density occurred in the interatrial septum (16.3+/-4.0 neurons/mm2). No neuronal somata expressed TH-IR or contained detectable amines but these elements were expressed by somata of small cells (mean total 124+/-33) throughout the atria, primarily associated with ganglia. Amine- and TH- containing varicosities were also present in ganglia, representing potential sites for adrenergic modulation of ganglionic neurotransmission. Branches of extrinsic cardiopulmonary and vagus nerves were distributed to all parts of both atria. The organisation of the intracardiac nervous system revealed in this study will facilitate further investigations of regional autonomic control of the heart.

Animals↗

Development of catecholaminergic neurons in the pond snail, Lymnaea stagnalis: I. Embryonic development of dopamine-containing neurons and dopamine-dependent behaviors.

The embryonic development of the catecholaminergic system of the pond snail, Lymnaea stagnalis, was investigated by using chromatographic and histochemical methods. High performance liquid chromatography suggested that dopamine was the only catecholamine present in significant concentrations throughout the embryonic development of Lymnaea. Dopamine first became detectable at about embryonic stage (E) 15 (15% of embryonic development) and then increased in amount during early development to reach about 120-140 fmol per animal by around E40. Dopamine content remained stable during mid-embryogenesis (E40-65), increased slowing for the next couple of days, and then increased rapidly to culminate at about 400 fmol per animal by hatching. The detection of aldehyde- and glyoxylate-induced fluorescence and of tyrosine hydroxylaselike immunoreactivity indicated that the first catecholaminergic cells appeared in the late trochophore or early veliger stage of embryonic development (E32-35). The paired perikarya of these transient apical catecholaminergic (TAC) neurons were located beneath the apical plate, remained outside of the central ganglia during embryogenesis, and no longer contained detectable catecholamines close to hatching. TAC neurons bore cilia on the ends of short processes that penetrated the overlying epithelium; their long processes branched repeatedly under the ciliated apical plate. Several smaller catecholaminergic cells first appeared in the anterior margin of the foot at a stage when the embryos began to metamorphose from the veliger form (E55). Similar bipolar cells later appeared in the tentacle and lips. The axons of all of these small peripheral cells projected centrally and terminated within the neuropil of different central ganglia. Central catecholaminergic neurons, including RPeD1, differentiated only after metamorphosis was complete (E75). Development of locomotor, respiratory, and feeding behaviors correlated with maturation of catecholaminergic neurons, as indicated by histology and chromatography.

Animals↗

Development of catecholaminergic neurons in the pond snail, Lymnaea stagnalis: II. Postembryonic development of central and peripheral cells.

Catecholamines have long been thought to play important roles in different mollusc neural functions. The present study used glyoxylate- and aldehyde-induced histofluorescence to identify central and peripheral catecholaminergic neurons in the snail Lymnaea stagnalis. The majority of these cells were also found to react to antibodies raised against tyrosine hydroxylase. A minority of the catecholaminergic neurons, however, exhibited no such immunoreactivity. The number of central catecholaminergic neurons nearly doubled (from about 45 to about 80 cells) during the first 2-3 days of postembryonic development. Thereafter, catecholaminergic neurons again doubled in number and generally grew by about 100-200% in soma diameter as the snails grew by 1,000% in overall linear measurements. In contrast to the relatively meager addition of central catecholaminergic neurons, several thousand catecholaminergic somata were added to different peripheral tissues during postembryonic development. These small, centrally projecting neurons were particularly concentrated in the lips, esophagus, anterior margin of the foot, and different regions of the male and female reproductive tracts. Chromatographic analyses indicated that dopamine was the major catecholamine present in the central ganglia, foot, and esophagus, although detectable levels of norepinephrine (approximately 20% of dopamine levels) were also found in the ganglia. The total content but not the concentration of dopamine increased within the tissue samples during postembryonic development. The companion study (Voronezhskaya et al. [1999] J. Comp. Neurol. 404:285-296) and the present study furnish a complete description of central and peripheral catecholaminergic neurons from their first appearance in early embryonic development to adulthood.

Animals↗

Serotonergic responses of the siphons and adjacent mantle tissue of the zebra mussel, Dreissena polymorpha.

Bivalve siphons have important functional roles in nutritional physiology, defense, and reproductive mechanisms, yet little is known about their neuromuscular control. In the present study, tension measurements and video observations of siphons and adjacent mantle tissue were used to investigate responses to serotonin (5-HT). 5-HT caused relaxation at 10(-6) M and 10(-5) M, contraction or biphasic responses at 10(-4) M and 10(-3) M, and siphon opening at 10(-4) M and 10(-3) M. Responses were slow and lasted 5-10 min after much shorter (20 s) applications of 5-HT. The relaxation phase was enhanced in high potassium medium. Contractile responses could be mimicked by alpha-1-methyl-5-HT and 2-methyl-5-HT but not by 8-OH-DPAT. The responses were not affected by methiothepin, TFMPP, 1-(1-naphthyl)piperazine, metergoline, NAN-190, mianserin, cyproheptadine, and ketanserin. The pharmacology of the 5-HT receptor(s) mediating these siphon/mantle responses is, therefore, different from previously described 5-HT receptors involved in spawning. The siphon/mantle contains previously undescribed longitudinal muscle fibers in the mantle and circular muscle fibers around the siphons. Serotonergic innervation of the siphon margins and mantle tissue was observed by immunohistology. The presence of 5-HT in the siphon/mantle tissue and the responsiveness of these preparations to 5-HT suggest that 5-HT may be a physiological regulator of mantle and siphon movements in the zebra mussel.

Animals↗

Pharmacological analysis of monoamine synthesis and catabolism in the scallop, Placopecten magellanicus.

1. The efficacies of various agents that affect monoamine synthesis, oxidation and methylation were evaluated in the scallop, Placopecten magellanicus, through the use of high performance liquid chromatography with electrochemical detection. 2. Central ganglia, labial palps and feet from animals bathed in 10(-5) M or 10(-4) M alpha-methyl-p-tyrosine for 1 day followed by 3-6 day recovery in artificial sea water had significantly reduced concentrations of 3,4-dihydroxyphenylalanine, norepinephrine, epinephrine, dopamine and 3,4-dihydroxyphenylacetic acid. 3. Central ganglia, labial palps and feet from scallops incubated in 10(-5) M or 10(-4) M para-chlorophenylalanine for 1 day followed by a 3-6 day wash in artificial sea water had significantly reduced concentrations of 5-hydroxytryptophan, 5-hydroxytryptamine and 5-hydroxy-3-indoleacetic acid. 4. Monamine oxidase inhibitors (administered at 10(-4) M for 1 day followed by a 2-day recovery) significantly decreased the concentrations of 3,4-dihydroxyphenylalanine and 5-hydroxy-3-indolacetic acid and increased the concentrations of their corresponding precursors in tissues. Deprenyl, a monoamine oxidase type B inhibitor, generally had more potent effects than pargyline, which inhibits monoamine oxidase type B and type A. Clorgyline, a monoamine oxidase type A specific inhibitor, showed few significant effects on the levels of the monoamines or their precursors or metabolites. 5. Bath application of 10(-4) M 3,5-dinitrocatechol, a blocker of catechol-O-methyl transferase, resulted in significant decreases in the concentrations of normetanephrine and metanephrine in nervous and other tissues and increased the levels of their corresponding precursors, dopamine, norepinephrine and epinephrine. 6. Generally, treatments that appeared to directly cause changes in levels of catecholaminergic compounds indirectly resulted in inverse changes in levels of indolaminergic compounds, and vice versa. This suggests an interaction between these transmitter systems. 7. The detection of monoaminergic compounds and dramatic changes in their concentrations following various drug effects strongly suggests the presence of mammalian-type metabolic pathways leading to synthesis and subsequent inactivation of monoamines.

Animals↗

Reproduction-associated immunoreactive peptides in the nervous systems of prosobranch gastropods.

Antibodies against reproductive peptides of Aplysia and Lymnaea were used to localize homologous immunoreactive peptides in the nervous systems of three prosobranch species: Busycon canaliculatum, Concholepas concholepas, and Tegula atra. Positive control experiments in L. stagnalis demonstrated the broad species range of the anti-egg-laying hormone (anti-ELH) antibody used in this study, and showed binding of anti-alpha-caudodorsal-cell peptide (anti-alpha-CDCP) to the same cells in cerebral and buccal ganglia. Dot immunoassays with synthetic ELH confirmed the reactivity and sensitivity (< 0.1 microgram) of the anti-ELH antibody. Experiments with preadsorbed antibody or no primary antibody confirmed its specificity. In B. canaliculatum, clusters of more than 300 neuronal cell bodies immunoreactive to both anti-ELH and anti-alpha-CDCP were observed along the medial margins of left and right cerebral ganglia. Anti-alpha-CDCP reacted with additional small populations of cerebral ganglion neurons not stained by anti-ELH. Anti-ELH and anti-alpha-CDCP also reacted with overlapping but different small populations of neurons in buccal ganglia. In C. concholepas and T atra, ELH-like immunoreactivity was found in cerebral ganglia, and in T. atra in fibers in the cerebral ganglia and cerebral-pedal connectives. Thus, cerebral ganglia are the major locus of the ELH-like immunoreactivity in prosobranchs.

Amino Acid Sequence↗

Molecular cloning of a cDNA encoding the neuropeptides APGWamide and cerebral peptide 1: localization of APGWamide-like immunoreactivity in the central nervous system and male reproductive organs of Aplysia.

While much is known about the neural and endocrine mechanisms that control egg laying in the gastropod mollusk Aplysia, relatively little is known about the regulation of male reproductive activity in this simultaneous hermaphrodite. In the present study, we have cloned and sequenced a cDNA that encodes a precursor protein, the predicted posttranslational processing of which presumably generates nine copies of the neuropeptide Ala-Pro-Gly-Trp-NH2 (APGWamide), five connecting peptide sequences, and a C-terminal peptide. The sequence of one connecting peptide is identical to the previously characterized cerebral peptide 1. Northern blot analysis identified two major APGWamide mRNA transcripts (approximately 1.3 kb, approximately 2.4 kb), which were present in central nervous system ganglia, but were most abundant in the right cerebral and right pedal ganglia. Immunohistochemical studies using sexually mature Aplysia demonstrated that the vast majority of APGWamide-like immunoreactivity was localized in 30-40 neurons along the anterior and medial margins of the right cerebral ganglion and in a cluster of 15-20 neurons in the right pedal ganglion. A total of only about ten immunoreactive neurons were located in other ganglia. Immunohistochemistry also demonstrated that APGWamide was present in the reproductive organs that participate in the storage or transport of sperm, including the small hermaphroditic duct (site of sperm storage before mating), the white hemiduct (also known as the copulatory duct), and penial complex. As a group, these data suggest that APGWamide may play a role in regulating male reproductive function in Aplysia, as it does in other gastropods.

Amino Acid Sequence↗

Characterization of an identified cerebrobuccal neuron containing the neuropeptide APGWamide (Ala-Pro-Gly-Trp-NH2) in the snail Lymnaea stagnalis.

A bilaterally symmetrical pair of cerebrobuccal neurons in Lymnaea stagnalis shows immunoreactivity for the molluscan neuropeptide APGWamide. The neuron somata are whitish in colour and located on the ventral surface of each cerebral ganglion between the roots of the labial nerves. A single axon travels via the ipsilateral cerebrobuccal connective into the buccal ganglia, where it gives rise to fine neuritic branching. Based upon these characteristics, the neuron has been named the cerebrobuccal white cell (CBWC). In isolated CNS preparations, in the absence of feeding motor output, the CBWC is silent and receives few, low amplitude, synaptic inputs. During generation of fictive feeding, the CBWC bursts in phase with cycles of feeding motor output. Tonic or phasic stimulation of CBWC leads to initiation of rhythmic feeding motor output. However, evoked bursts of activity in CBWC, which mimic its normal burst pattern, cannot entrain the buccal rhythm, suggesting that CBWC is not itself a major component of the feeding central pattern generator (CPG). Strong stimulation of CBWC during ongoing feeding motor output leads to a reduction in frequency and/or intensity of the buccal rhythm. Bath application of synthetic APGWamide (10(-7)M-10(-4)M) to the isolated CNS can activate feeding motor output in quiescent preparations after a delay, but disrupts ongoing buccal rhythms. This study represents the first description of a peptidergic cerebrobuccal neuron in the well described gastropod feeding system and also provides new information about the role of a novel molluscan neuropeptide.

Action Potentials↗

Detection of APGWamide-like immunoreactivity in the sea scallop, Placopecten magellanicus.

In pulmonate gastropods, the peptide, Ala-Pro-Gly-Trp-NH2 (APGWamide), appears to be located nearly exclusively in the neural circuitry controlling the male reproductive organs. This neuropeptide and related neuropeptides are also present and apparently bioactive in bivalve molluscs, although their physiological role in these latter animals is unknown. The present report uses immunohistochemistry to examine the distribution of APGWamide and/or related peptides within the tissues of the deep sea scallop, Placopecten magellanicus. Much of the APGWamide-like immunoreactivity (APGWa-LIR) was detected in the central nervous system (CNS) of both juvenile and adult scallops, where it was concentrated in the cerebral, pedal and parietovisceral ganglia, particularly in the cortex of ganglionic cells and their axons which extend into the central neuropilar region. APGWa-LIR was also detected in the nerves ramifying from these ganglia. In addition, strong APGWa-LIR was localized in what appeared to be axonal terminals within peripheral tissues including the striated adductor muscle, foot, gills, labial palps, lips, tentacles and gonads of the juvenile scallops. The presence of APGWa-LIR was also confirmed in the gonads of adults of both sexes. Cursory examinations of the CNS of the mussel, Mytilus edulis, and the oyster, Crassostrea virginica, revealed the presence of APGWa-LIR in cell bodies and processes in these bivalve species as well. It is concluded that APGWamide and/or related peptides are probably important neurotransmitters and/or neuromodulators of several central and peripheral functions in P. magellanicus and other bivalves. Future work must focus on the possible roles for APGWamide in the physiological processes of these and other bivalve species.

Aging↗

Serotonin depletion after prolonged chlorpromazine treatment in a simpler model system.

1. Prolonged exposure of the pond snail Lymnaea stagnalis to micromolar concentrations of chlorpromazine (CPZ) results in marked changes in the serotonin (5-HT) content of the central nervous system. 2. High-performance liquid chromatography with electrochemical detection indicates that levels of 5-HT, but not those of dihydroxyphenyl-alanine (DOPA), dopamine or norepinephrine, were significantly decreased (e.g., to less than 40% of normal after 30 days of exposure to 1 microM CPZ in the bathing water). 3. Glyoxylate-induced fluorescence was depressed to undetectable levels in central, serotonergic neurons. 4. Performance of 5-HT-dependent motor behaviors was impaired. 5. The present results, in accord with earlier studies on the effects of chronic exposure to haloperidol, suggest that previously overlooked mechanisms of monoamine downregulation may contribute to long-term effects of antipsychotic drugs.

Animals↗

Cholinergic and peptidergic regulation of siphon/mantle function in the zebra mussel, Dreissena polymorpha.

Neurotransmitter regulation of the siphon and adjacent mantle region of bivalves has not previously been examined. In the biofouling bivalve, Dreissena polymorpha, acetylcholine and FMRFamide both elicited contractions of siphon/mantle preparations. Hexamethonium bromide inhibited acetylcholine-elicited contractions but had no effect on FMRFamide-elicited contractions. FMRFamide-like immunoreactivity and chromatographic evidence for acetylcholine were found in central ganglia and the siphon/mantle region. Extracts of siphons, gonads, and gills, separated on Sephadex G-25, also contained macromolecules larger than acetylcholine and FMRFamide that caused siphon/mantle contraction. These results demonstrate regulation of contraction by several potential neurotransmitter agents in a new bivalve preparation, the siphon/mantle.

Acetylcholine↗

Functional role of peptidergic anterior lobe neurons in male sexual behavior of the snail Lymnaea stagnalis.

A morphologically defined group of peptidergic neurons in the CNS of the hermaphroditic snail, Lymnaea stagnalis, is concerned with the control of a very specific element of male sexual behavior. These neurons are located in the anterior lobe of the right cerebral ganglion (rAL). By using chronically implanted electrodes, we show that the rAL neurons are selectively active during eversion of the penis-carrying structure, the preputium. The preputium is normally contained inside the body cavity and is everted during copulation in the male role. Electrical stimulation of the rAL neurons through the implanted electrodes, induced eversion of the preputium in vivo. Injection of APGWamide (Ala-Pro-Gly-Try-NH2), a small neuropeptide that is present in all rAL neurons, induced eversion of the preputium. Application of APGWamide to in vitro preparations of the preputium caused relaxation of this organ. In contrast, injection of the neuropeptide conopressin, which is co-localized with APGWamide in 60% of the rAL neurons, did not induce any behavior associated with male sexual activities. These results show that the neurons of the rAL can induce an eversion of the preputium as occurs during male copulation by release of APGWamide during a period of electrical activity.

Animals↗