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PubMed1996

The peptidergic nervous system of the triclad turbellarian, Bdelloura candida (Maricola, Bdellouridae): an immunocytochemical study using an antiserum raised to an endogenous neuropeptide, GYIRFamide.

The organisation of the nervous system of Bdelloura candida (Tricladida, Maricola, and Bdellouridae) was studied by immunocytochemistry, by using an antiserum raised to the authentic B. candida FMRFamide-related peptide (FaRP), GYIRFamide. Immunostaining was intense and abundant throughout both the central and peripheral nervous systems, being localised to the brain, the longitudinal nerve cords and their transverse and lateral connections, the pharyngeal plexus, the extensive sub-epidermal and sub-muscular plexuses, and elements of the reproductive apparatus. Compared to an earlier anatomical investigation of this species, and also to the neuroanatomy of other triclad turbellarians, the pattern of GYIRFamide-immunoreactivity reveals differences in the following aspects: the shape and structure of the brain, the distribution of longitudinal nerve cords and their relationships with the peripheral nervous system, the structure and distribution of the lateral nerves and the transverse connectives between the longitudinal nerve cords, organisation of the pharyngeal nervous system, and innervation of the eyespots and epidermal sensory structures. Although this study focuses on a descriptive account of the neuroanatomy of Bdelloura candida, by using anti-GYIRFamide as a neuronal marker, the possible functions of the native peptide are also discussed. The quality and reproducibility of the immunostaining obtained during this work highlights the effectiveness of the GYIRFamide antiserum in the neuroanatomical study of flatworms, and also the suitability of B. candida as a model species in studies of the turbellarian nervous system.

Animals↗
PubMed2003

Neurosecretory identity conferred by the apterous gene: lateral horn leucokinin neurons in Drosophila.

The LIM-HD protein Apterous has been shown to regulate expression of the FMRFamide neuropeptide in Drosophila neurons (Benveniste et al. [1998] Development 125:4757-4765). To test whether Apterous has a broader role in controlling neurosecretory identity, we analyzed the expression of several neuropeptides in apterous (ap) mutants. We show that Apterous is necessary for expression of the Leucokinin neuropeptide in a pair of brain neurons located in the lateral horn region of the protocerebrum (LHLK neurons). ap null mutants are depleted of Leucokinin in these cells, whereas hypomorphic mutants show reduced Leucokinin expression. Other Leucokinin-containing neurons are not affected by mutations in ap gene. Co-expression of apterous and Leucokinin is observed exclusively in the LHLK neurons, from larval stages to adulthood. Rescue assays performed in null ap mutants, by expressing Apterous protein under apGAL4 and elavGAL4 drivers, demonstrate the recovery of Leucokinin in the LHLK neurons. These results reinforce the emerging role of the LIM-HD proteins in determining neuronal identity. They also clarify the neuroendocrine phenotype of apterous mutants.

Animals↗
PubMed2003

Development of the larval nervous system of the gastropod Ilyanassa obsoleta.

Gastropods have been well studied in terms of early cell cleavage patterns and the neural basis of adult behaviors; however, much less is known about neural development in this taxon. Here we reveal a relatively sophisticated larval nervous system in a well-studied gastropod, Ilyanassa obsoleta. The present study employed immunocytochemical and histofluorescent techniques combined with confocal microscopy to examine the development of cells containing monoamines (serotonin and catecholamine), neuropeptides (FMRFamide and leu-enkephalin related peptides), and a substance(s) reactive to antibodies raised against dopamine beta-hydroxylase. Neurons were first observed in the apical organ and posterior regions during the embryonic trochophore stage. During later embryonic development neurons appeared in peripheral regions such as the foot, velum, and mantle and in the developing ganglia destined to become the adult central nervous system. In subsequent free-swimming veliger stages the larval nervous system became increasingly elaborate and by late larval stages there existed approximately 26-28 apical cells, 80-100 neurons in the central ganglia, and 200-300 peripherally located neurons. During metamorphosis some populations of neurons in the apical organ and in the periphery disappeared, while others were incorporated into the juvenile nervous system. Comparisons of neural elements in other molluscan larvae reveal several similarities such as comparable arrangements of cells in the apical organ and patterns of peripheral cells. This investigation reveals the most extensive larval nervous system described in any mollusc to date and information from this study will be useful for future experimental studies determining the role of larval neurons and investigations of the cellular and molecular mechanisms governing neural development in this taxon.

Animals↗
PubMed2001

Taurine-, aspartate- and glutamate-like immunoreactivity identifies chemically distinct subdivisions of Kenyon cells in the cockroach mushroom body.

The lobes of the mushroom bodies of the cockroach Periplaneta americana consist of longitudinal modules called laminae. These comprise repeating arrangements of Kenyon cell axons, which like their dendrites and perikarya have an affinity to one of three antisera: to taurine, aspartate, or glutamate. Taurine-immunopositive laminae alternate with immunonegative ones. Aspartate-immunopositive Kenyon cell axons are distributed across the lobes. However, smaller leaf-like ensembles of axons that reveal particularly high affinities to anti-aspartate are embedded within taurine-positive laminae and occur in the immunonegative laminae between them. Together, these arrangements reveal a complex architecture of repeating subunits whose different levels of immunoreactivity correspond to broader immunoreactive layers identified by sera against the neuromodulator FMRFamide. Throughout development and in the adult, the most posterior lamina is glutamate immunopositive. Its axons arise from the most recently born Kenyon cells that in the adult retain their juvenile character, sending a dense system of collaterals to the front of the lobes. Glutamate-positive processes intersect aspartate- and taurine-immunopositive laminae and are disposed such that they might play important roles in synaptogenesis or synapse modification. Glutamate immunoreactivity is not seen in older, mature axons, indicating that Kenyon cells show plasticity of neurotransmitter phenotype during development. Aspartate may be a universal transmitter substance throughout the lobes. High levels of taurine immunoreactivity occur in broad laminae containing the high concentrations of synaptic vesicles.

Aging↗
PubMed2004

Antho-RFamide-containing neurons in the primitive nervous system of the anthozoan Renilla koellikeri.

The neuropeptide Antho-RFamide is extremely abundant in Renilla koellikeri (sea pansy), a representative of the cnidarians (octocorallians) considered to be closest to the stem ancestors of metazoans with nervous systems. Therefore, a knowledge of the distribution of Antho-RFamide-containing neurons in this species would contribute to our understanding of the early evolution of nervous systems. Using antisera raised against RFamide and FMRFamide, we detected immunostaining in numerous neurons throughout the nervous system of the sea pansy. The antisera revealed ectodermal nerve-nets on the upper and lower sides of the colony and on the oral side of tentacles, in the oral disk, and in the pharynx of feeding polyps. Neurons were immunostained also in the mesogleal nerve-net of feeding polyps and in the through-conducting mesogleal nerve-net of the colonial mass. Varying densities of stained neurons were observed in the different compartments of the endoderm: muscular walls of the feeding and water circulation polyps, mesenteric filaments and their derived follicles containing either ovocytes or spermatophores, in the endodermal channels connecting the different compartments of the colony, and in circular muscle of the peduncle. The distribution of immunostained neurons suggests that they play important roles in feeding, reproduction, neuromuscular transmission, and in neuro-neuronal transmission coordinating the different parts of the colony.

Animals↗
PubMed2004

Peptidomics of CNS-associated neurohemal systems of adult Drosophila melanogaster: a mass spectrometric survey of peptides from individual flies.

Neuropeptides are important messenger molecules that influence nearly all physiological processes. In insects, they can be released as neuromodulators within the central nervous system (CNS) or as neurohormones into the hemolymph. We analyzed the peptidome of neurohormonal release sites and associated secretory peptidergic neurons of adult Drosophila melanogaster. MALDI-TOF mass spectrometric analyzes were performed on single organs or cell cluster from individual flies. This first peptidomic characterization in adult fruit flies revealed 32 different neuropeptides. Peptides not directly predictable from previously cloned or annotated precursor genes were sequenced by tandem mass spectrometry. These peptides turned out to be either intermediate products of neuropeptide processing or shorter versions of known peptides. We found that the peptidome of the CNS-associated neurohemal organs is tagma-specific in Drosophila. Abdominal neurohemal organs and their supplying peptidergic neurons contain the capa gene products periviscerokinins and pyrokinin-1, thoracic neurohemal organs contain FMRFamides, and the neurohemal release sites of the brain contain pyrokinin-1(2-15), pyrokinin-2, corazonin, myosuppressin, and sNPF as their major putative release products. Our results show that peptidomic approaches are well suited to study differential neuropeptide expression or posttranslational modifications in morphologically defined parts of the nervous system and in a developmental and physiological context in animals as small as Drosophila melanogaster.

Animals↗
PubMed1988

Immunocytochemical and morphological evidence for a retinopetal projection in anuran amphibians.

The centrifugal projection to the retina in anuran amphibians (Rana catesbeiana and Xenopus laevis) has been investigated by immunocytochemistry, HRP transport, and optic nerve lesionings. FMRFamide- and N-terminal substance P-immunoreactive (Fa-ir and SP(3-7)-ir) fibers were abundant in the normal retina and optic nerve but almost absent distal to an optic nerve section or crush after 7-14 days survival. Fa-ir and SP(3-7)-ir fibers were traced to the optic nerve from the lamina terminalis (or the septopreoptic junctional area), where there are many Fa-ir and SP(3-7)-ir perikarya. After application of HRP to the optic nerve and survival for 9-10 days, retrogradely labeled neurons were observed in the lamina terminalis. Conversely, following HRP injection into the septal and preoptic area, labeled fibers were observed in the optic nerve. These results suggest that Fa-ir and SP(3-7)-ir efferent fibers project from the lamina terminalis to the retina. But in anurans, unlike teleosts, these fibers are not gonadotropin-releasing-hormone (GnRH)-ir. The morphological relations of this retinopetal pathway with the GnRH-ir nervus terminalis are discussed.

Animals↗
PubMed1991

Peptide-immunocytochemistry of neurosecretory cells in the brain and retrocerebral complex of the sphinx moth Manduca sexta.

Antisera against a variety of vertebrate and invertebrate neuropeptides were used to map cerebral neurosecretory cells in the sphinx moth Manduca sexta. Intense immunoreactive staining of distinct populations of neurosecretory cells was obtained with antisera against locust adipokinetic hormone, bovine pancreatic polypeptide, FMRFamide, molluscan small cardioactive peptide (SCPB), leucine-enkephalin, gastrin/cholecystokinin, and crustacean beta-pigment dispersing hormone (beta PDH). Other antisera revealed moderate to weak staining. Each type of neurosecretory cell is immunoreactive with at least one of the antisera tested, and most of these neurons can be identified anatomically. The staining patterns provide additional information on the organization of cerebral neurosecretory cells in M. sexta. Based upon anatomical and immunocytochemical characteristics, 11 types of neurosecretory cells have been recognized in the brain, one type in the suboesophageal ganglion, and one in the corpus cardiacum. Extensive colocalization experiments show that many neurosecretory cells are immunoreactive with several different antisera. This raises the possibility that these cells may release mixtures of neuropeptides into the hemolymph, as has been demonstrated in certain other systems. The immunocytochemical data should be helpful in efforts to identify additional peptide neurohormones released from the brain of this and other insects.

Animals↗
PubMed1991

Ontogeny of the F8Famide-like (morphine-modulating) peptides in the central nervous system of rats.

FLFQPQRF-NH2 (F8Famide; FMRFamide-like peptide; morphine-modulating peptide) is a peptide isolated from bovine brain. It has some opiate analgesia modulating effects. In an adult rat central nervous system, F8Famide-like immunoreactivity is found in high concentrations in the posterior pituitary, hypothalamus, pons, medulla, and dorsal spinal cord. By using immunohistochemistry and radioimmunoassay, we studied the development of the F8Famide-immunoreactive system in the rat central nervous system during the ontogeny. F8Famide-immoreactive fibers and terminals first appeared in the median eminence on the twentieth embryonal day. Postnatally the F8Famide-immunoreactive system developed rapidly both in the brain and spinal cord, the immunoreactive structures having an adultlike distribution by the age of 1 week. A transient increase of the F8Famide-immunoreactive material was observed during the third and fourth postnatal weeks. By the age of 4 weeks, the F8Famide-containing neuronal system was adultlike both in distribution and quantity. The results suggest that the F8Famide-like peptides may act as neurotransmitters or neuromodulators from the time of their appearance in the brain and spinal cord. The early appearance and the distribution of the F8Famide-like immunoreactivity suggest that these peptides may participate, in addition to nociceptive mechanisms, in the regulation of blood pressure, feeding behaviour, and endocrine functions.

Amino Acid Sequence↗
PubMed1991

Galanin immunoreactivity in the blowfly nervous system: localization and chromatographic analysis.

In this study chromatographic, immunochemical, and immunocytochemical methods provide evidence of a galanin-like peptide(s) in an invertebrate, the blowfly Phormia terraenovae. The major portion of the galanin-like immunoreactivity (GAL-LI) in fly heads was extractable in acetic acid but not in boiling water, which suggests that the peptide(s) may be highly basic in nature. GAL-LI was present both in the head and body portion of the blowfly in roughly the same amounts. Initial gel filtration data, using a G-50 Sephadex column and a weak phosphate-buffer (pH 6.5) as eluent, suggested that a fly GAL-LI peptide(s) from fly heads, eluting as an apparent single peak, was smaller than porcine GAL(1-29) and GAL(1-15). However, concomitant analysis using a G-25 Sephadex column and acetic acid (0.2 M) as eluent, spread the immunoreactive material over a great portion of the chromatogram, although the main portion of the material eluted in the same size range as porcine GAL(1-29). Taken together, the gel filtration data thus suggest that fly GAL-LI peptide(s) may be highly basic but presumably similar in size to vertebrate GAL(1-29). However, the hydrophobic properties of the fly GAL-LI peptide(s) differ from that of porcine GAL as demonstrated by the presence of several immunoreactive components eluting both early as well as late in the chromatogram when using reverse-phase high performance liquid chromatography (HPLC); early peaks may represent highly basic and/or possibly smaller GAL-immunoreactive peptide(s), whereas later peaks may represent less basic and possibly elongated forms. Immunocytochemistry indicated that GAL-LI was present in the nervous system of the blowfly. About 160 GAL-immunoreactive neurons were found in the brain and subesophageal ganglion, 26 in the fused thoracic ganglion and 30 in the fused abdominal ganglion. In the brain, GAL-immunoreactive fibers supply specific subdivisions of the central body, optic lobe, superior protocerebrum, and tritocerebrum as well as neuropil in the subesophageal ganglia. In the thoracico-abdominal ganglia, GAL-immunoreactive neuron processes are found inside synaptic neuropil as well as in the neural sheath of the ganglia and several of the dorsal nerve roots. Many of the GAL-immunoreactive neurons react also with an antiserum against porcine galanin message associated peptide, a peptide present in the preprogalanin protein. Immunocytochemical double-labeling indicated that some GAL-immunoreactive neurons also reacted with antisera against the molluscan peptides FMRFamide and SCPB, whereas no evidence could be found for colabeling with antisera against tyrosine hydroxylase, substance P and physalaemin.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Pigment-dispersing hormone-like peptide in the nervous system of the flies Phormia and Drosophila: immunocytochemistry and partial characterization.

beta-pigment-dispersing hormone (beta-PDH) isolated from the fiddler crab (Rao et al., '85) is a member of an octadecapeptide family of neuropeptides common to arthropods. Whereas earlier studies of these peptides in insects were limited to orthopterans, this investigation focuses on dipteran flies. Extracts of heads from the blowfly Phormia terraenovae were assessed in a fiddler crab bioassay for PDH activity. Immunocytochemistry, dose-response curves, gel filtration chromatography and reversed-phase HPLC, combined with bioassay and enzyme-linked immunosorbent assay (ELISA), indicate the presence of PDH-like peptide in the blowfly. Immunocytochemical mapping of PDH-like immunoreactive (PDHLI) neurons was performed for the entire nervous systems of Phormia and the fruitfly Drosophila with a beta-PDH antiserum. In the cephalic ganglion (brain, optic lobe and subesophageal ganglion) PDHLI cell bodies could be detected (34 in Phormia and 16 in Drosophila). In both species, each hemisphere contains 8 PDHLI cell bodies in the optic lobes. These innervate the optic lobe neuropils bilaterally. In Phormia, another set of 8 cell bodies are located in each of the lateral neurosecretory cell groups in the superior protocerebrum. These neurons send axons to the corpora cardiaca-hypocerebral ganglion complex and to portions of the foregut. In contrast, only the optic lobe neurons display immunoreactivity in Drosophila. Except for the optic lobes, PDHLI processes are distributed only in nonglomerular neurophils of the brain of both species. In the fused thoracico-abdominal ganglia of Phormia, 28 PDHLI cell bodies were found (only six were found in Drosophila). In both species, six abdominal PDHLI neurons are efferents with axons innervating the hindgut. We also found that some of the PDHLI neurons in the Phormia brain and abdominal ganglion contain colocalized FMRFamide-like immunoreactivity. Since the flies studied here do not display hormonally controlled, fast pigment migrations, the PDH-like peptide may have a role as neurotransmitter or neuromodulator in the central nervous system, especially in the visual system, and a regulatory role in the stomatogastric system and the hind-gut.

Animals↗
PubMed1993

Immunocytochemical localization of neuropeptide FF (FMRF amide-like peptide) in the hypothalamo-neurohypophyseal system of Wistar and Brattleboro rats by light and electron microscopy.

Neuropeptide FF (F8Famide, FMRFamide-like, or morphine modulating peptide) immunoreactivity was localized by light and electron microscopy in the hypothalamo-neurohypophyseal system of Wistar and Brattleboro rats. In Wistar rats neuropeptide FF was present in part of the magnocellular neurones of the paraventricular and supraoptic nuclei in which it was coexpressed with vasopressin. Neuropeptide FF containing fibres were present in the paraventricular and the supraoptic nuclei, and in the central part of the neural lobe. At the electron microscopic level, neuropeptide FF containing nerve terminals in the neural lobe formed synaptoid contacts exclusively with pituicytes. No neuropeptide FF containing neurovascular contacts or contacts with other neuronal structures were observed. In contrast with Wistar rats, neuropeptide FF was almost completely absent in cell bodies of the paraventricular and supraoptic nuclei, and in fibres of the neural lobe in Brattleboro rats. Only a few solitary cells could be observed in these structures. The present results demonstrate that neuropeptide FF coexists with vasopressin within the hypothalamo-neurohypophyseal system. As we did not observe neuropeptide FF containing neurovascular contacts, neuropeptide FF containing nerve terminals probably have a local function within the neural lobe. Neuropeptide FF may be involved in the modulation of oxytocin and vasopressin release, with the pituicyte as an intermediate cell.

Animals↗
PubMed1995

Immunocytochemical mapping of serotonin and neuropeptides in the accessory medulla of the locust, Schistocerca gregaria.

Accumulating evidence suggests that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla of the insect brain are involved in circadian pacemaking functions. We have used immunocytochemical techniques to investigate the neurochemical organization of the accessory medulla in the locust Schistocerca gregaria. Local neurons with arborizations largely restricted to the accessory medulla are immunoreactive with antisera against serotonin, Manduca sexta allatotropin, and Diploptera punctata allatostatin 7. Projection neurons with arborizations in the accessory medulla and fibers to the lamina and/or several areas in the midbrain including the posterior optic tubercles, the inferior and the superior protocerebrum show Phe-Met-Arg-Phe (FMRF)amide-, gastrin/cholecystokinin-, crustacean cardioactive peptide-, and substance P immunoreactivities. A unique neuron with tangential ramifications in the medulla and lamina and varicose terminals in the accessory medulla contains a peptide related to locustatachykinin I/II. Double-label experiments show colocalization of pigment-dispersing hormone-immunoreactivity with substances related to gastrin/cholecystokinin, FMRFamide, substance P, or crustacean cardioactive peptide in certain projection neurons of the accessory medulla. The results suggest that neuropeptides and biogenic amines play major neuroactive roles in the accessory medulla of the locust. The abundance and extensive colocalization of neuropeptides in the locust accessory medulla is discussed with respect to the possible involvement of this brain area in circadian pacemaking functions.

Animals↗
PubMed1984

Gastrointestinal hormones in birds: morphological, chemical, and developmental aspects.

Historically, the enterochromaffin cell was the first endocrine cell type detected in avian gut; subsequently, a number of types of such cells were distinguished on the basis of the ultrastructural features of the secretory granules. More recently, immunocytochemical procedures have revealed somatostatin-, pancreatic polypeptide (PP)-, polypeptide YY-, glucagon-, secretin-, vasoactive intestinal peptide (VIP)-, gastrin-, cholecystokinin-, neurotensin-, bombesin-, substance P-, enkephalin-, motilin-, and FMRFamide-like immunoreactivity in avian gastrointestinal endocrine cells. Most endocrine cells are located in the antrum; there are a number in the proventriculus and small intestine but few in the gizzard, cecum, and rectum. Several avian gastroenteropancreatic hormones, including glucagon, VIP, secretin, bombesin, neurotensin, and PP, have been isolated and sequenced. They resemble the equivalent mammalian peptides in terms of molecular size but differ in amino acid composition and sequence; some (e.g., VIP) differ only in minor respects, others (e.g., secretin) more radically. Gastrointestinal endocrine cells appear late in development; available data indicate that few types are recognized by either immunocytochemistry or electron microscopy before 16 days of incubation. Experimental evidence has shown that at least the majority of gut endocrine cells are of endodermal origin and are not derived from the neural crest or neuroectoderm as earlier proposed. In early embryos, the progenitors of gastrointestinal endocrine cells are more widespread than are the differentiated cells in chicks at hatching. This, along with other observations, raises the question of factors that might influence the differentiation of gut endocrine cells.

APUD Cells↗
PubMed1985

Cross-phyletic bioactivity of arthropod neurohormones and molluscan ganglion extracts: evidence of an extended peptide family.

Two structurally related arthropod neuropeptides, red pigment concentrating hormone (RPCH) and adipokinetic hormone (AKH), are potent excitors of the heart of the clam Mercenaria mercenaria. The response is bimodal: whereas the threshold for affected hearts is 1-3 X 10(-9) M, about 40% of the preparations are virtually unresponsive. Aqueous extracts of Mercenaria ganglia contain a substance which concentrates the red pigment in the erythrophores of intact destalked Uca pugilator and even of its isolated legs. This substance is retained on Sephadex G-15 and co-elutes with synthetic shrimp RPCH. The active fractions also concentrate the erythrophores and the leucophores of destalked shrimp (Penaeus). Neither dopamine nor the molluscan neuropeptide FMRFamide had any chromatophorotropic effect in these assays. The activity of the ganglion extracts was abolished by digestion with chymotrypsin. In conclusion, molluscan ganglion extracts contain a peptide factor, possibly an analog of RPCH, that concentrates the pigments of crustacean chromatophores by a direct action on the cells.

Animals↗
PubMed2005

Immunocytochemistry of the nervous system and the musculature of the chordoid larva of Symbion pandora (Cycliophora).

To date, the phylum Cycliophora comprises only one described extant species of acoelomate marine invertebrates, Symbion pandora. Adult specimens live commensally on the mouthparts of the Norwegian lobster, Nephrops norvegicus. Its complicated life cycle includes an asexually produced Pandora larva and a sexually produced chordoid larva. Despite detailed TEM investigations and its inclusion in recent molecular phylogenetic analyses, cycliophoran relationships still remain enigmatic. In order to increase the morphological database, I investigated the anatomy of the nervous system and the musculature of the chordoid larva by applying fluorescence-coupled antibodies against the neurotransmitters serotonin and FMRFamide, as well as FITC-coupled phalloidin to label filamentous F-actin, in combination with confocal laser scanning microscopy. The FMRFamidergic nervous system shows a bilobed anterior ganglion and one pair of ventral nerve cords, while serotonin is distributed in a scattered pattern in the anterior ganglion. In addition, there are two pairs of ventral serotonergic nerves, of which the inner pair fuses with the outer nerve cords in the posterior third of the larva. The musculature comprises an outer layer of six units of circular body wall muscles, several helicoid muscle fibers, a set of paired longitudinal muscles that span the entire anterior-posterior axis of the larva, and a few oblique muscle strands. Furthermore, an anterior muscle complex and one pair of posterior muscles are present. The chordoid organ consists of a number of distinct subunits that are each formed by a dense layer of circular muscle fibers. The overall arrangement of the oblique and longitudinal muscles as well as the body wall musculature in the chordoid larva of Symbion pandora exhibits similarities with the condition found in certain rotifers. This is congruent with some recent phylogenies based on 18S rRNA sequences but additional morphological, developmental, and molecular data are needed to clarify the phylogenetic relationships of Cycliophora.

Animals↗
PubMed1984

Cholinergic receptors in the Aplysia gill.

Acetylcholine has been suggested as a neurotransmitter released in the Aplysia gill by peripheral afferents of central neurons and by peripheral neurons within the gill. The perfused gill, isolated from the abdominal ganglion, was examined. At concentrations greater than 1 microM, acetylcholine elicited a slowly developing tonic contraction of the afferent vein that reversed upon washout. This effect was observed on both quiescent and active preparations. At concentrations less than 1 microM, acetylcholine perfusion resulted in a reduction of gill tone. The excitatory effect of acetylcholine was reduced 80 and 60% by the cholinergic antagonists atropine and hexamethonium, respectively. The acetylcholine-evoked contraction was potentiated 2.5-fold when curare was coinfused. Carbachol did not mimic the excitatory effects of acetylcholine. At all concentrations examined (1-100 microM), carbachol infusion reduced baseline tension, the amplitude of spontaneous contractions and contractions evoked by FMRFamide and dopamine. Contractions evoked by perfusion of p-chlorophenylthiocyclic AMP were greatly reduced when carbachol was added to the perfusate. Further addition of curare reversibly blocked carbachol inhibition of the cyclic AMP-evoked contractions. These findings suggest that excitatory and inhibitory cholinergic receptors are involved in the regulation of gill contractile behavior by acetylcholine.

Acetylcholine↗
PubMed1989

Catecholamine neurons in Aplysia: improved light-microscopic resolution and ultrastructural study using paraformaldehyde and glutaraldehyde (FaGlu) cytochemistry.

We have modified the formaldehyde-glutaraldehyde (FaGlu) histofluorescence method of Furness, Costa, and Blessing (1977a) and Furness, Costa, and Wilson (1977b) to examine wholemounts and sections of both juvenile and adult ganglia as well as peripheral tissues of Aplysia californica. FaGlu fluorescence is the result of a reaction between formaldehyde and tissue catecholamines to produce water-insoluble (fixed) fluorophores. In serially sectioned cerebral ganglia, 70-80 positive neurons were observed (many in clusters of 10-20 cells), many more than were found using the glyoxylic acid technique. Catecholamine-containing varicosities were densely packed in localized portions of the neuropil of all central ganglia. Exclusive localization in the neuropil of presumed dopamine release sites is similar to that previously found for the neuropeptide SCP but differs from the widespread ramification of varicose neurites containing 5-HT, FMRFamide, and ELH. The FaGlu technique also enabled us to study the ultrastructure of catecholamine-containing neurons. In contrast to the larger vesicles found in serotonergic and histaminergic neurons, these dopaminergic neurons contain 70 nm dense-cored vesicles.

Animals↗