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Comparative Distribution of Neurons Containing FLFQPQRFamide-like (morphine-modulating) Peptide and Related Neuropeptides in the Rat Brain.

FLFQPQRF-NH2 (F8Famide; morphine-modulating peptide), isolated from bovine brain, is an FMRFamide-like peptide with opioid analgesia modulating effects. In the rat brain, F8Famide is immunohistochemically localized in neurons of the medial hypothalamus and medulla oblongata. Neuropeptide Y (NPY) is structurally related to F8Famide and the mammalian FMRFamide-like immunoreactivity (LI) was once thought to be due to an NPY-like peptide. We compared the anatomical distribution of F8Famide-LI with the localization of enkephalin- and NPY-LI-containing structures in the rat brain to find out if NPY or enkephalins coexist with F8Famide-LI. Cryostat sections of colchicine-treated Wistar rat brains were incubated with specific antisera against F8Famide, NPY, YGGFMRGL (Met-enkephalin-Arg-Gly-Leu), or YGGFMRF (Met-enkephalin-Arg-Phe) raised in rabbits. The immunoreactivity was visualized by the peroxidase - antiperoxidase or immunofluorescence method. The light microscopic mirror method was applied to study the colocalization of F8Famide and NPY. The F8Famide-immunoreactivity was concentrated in smaller areas of medial hypothalamus and nucleus of the solitary tract than that of enkephalins and NPY. In all brain areas, the distributions of F8Famide-, enkephalin- and NPY-immunoreactive neurons were distinct. F8Famide-, NPY- and enkephalin-LI-containing nerve terminals were seen in the nucleus of the solitary tract and in the lateral parabrachial nucleus. These results show that the neuronal systems containing F8Famide-, enkephalin- or NPY-LI are anatomically separate in all brain regions. However, there are terminal areas in which more than one type of these immunoreactivities are detected. These results have anatomical correlation with pharmacological reports, suggesting modulatory functions for these peptides on regulation of blood pressure, feeding behaviour and endocrine functions.

Journal Article↗

Innervation of vascular and cardiac muscle of Aplysia by multimodal motoneuron L7.

1. The cardiovascular system of Aplysia has proven to be a useful preparation for study of the neural control of circulation. To better understand the neural integration of function in this system, we have attempted to gain a more complete picture of its morphology and innervation patterns, with particular emphasis on the abdominal aorta and heart. 2. The vasoconstrictor muscle fibers of the abdominal aorta were found by dye injection to be extensively branched, with many processes that are less than 1 micron in diameter. Because of the wide spacing between individual muscle fibers, these fine processes, which have relatively few contractile filaments, may be required to mediate the electrical coupling that is observed between muscle cells. 3. L7, an identified cell in the abdominal ganglion, had been shown by others to be an excitatory motoneuron for the gill, the siphon, and the sheath-contracting muscles of the pleuroabdominal connectives, and also to excite the gill motoneurons in the branchial ganglion (5, 43, 56). We have found that this multimodal motoneuron also directly excites the auricle of the heart and the vasoconstrictor muscle of the abdominal aorta. 4. The excitatory effect of L7 on the abdominal aorta interacts synergistically with that produced by the other known excitatory inputs to that structure, the LBVc vasoconstrictor motoneurons. 5. The abdominal aorta is also richly innervated by axons immunoreactive for serotonin and for the neuropeptide FMRFamide. Serotonin inhibits the contractions of the aorta elicited by firing either L7 or the LBVC cells. In contrast, FMRFamide selectively inhibits the contractions elicited by the LBVC cells. 6. Our results suggest that a significant amount of the functional integration of the cardiovascular and respiratory systems is achieved by the use of a motoneuron common to both systems, and that there is likely to be extensive peripheral inhibitory modulation of the vasoconstrictor inputs to the abdominal aorta.

Animals↗

Modulation of an inhibitory interneuron in the neural circuitry for the tail withdrawal reflex of Aplysia.

1. Serotonin (5-HT), small cardioactive peptide B (SCPB) and FMRFamide have well-established facilitatory and inhibitory effects on sensory neurons and their connections with motor neurons mediating withdrawal reflexes in Aplysia. Little is known, however, about their effects on interneurons contributing to those reflexes. As a first step, we examined the effects of these three transmitters on the identified inhibitory interneuron RP14 in isolated pleural-pedal ganglia. 2. Bath application of 5-HT hyperpolarized RP14, inhibited its spontaneous activity and decreased its excitability. In addition, 5-HT decreased the amplitude of inhibitory postsynaptic potentials produced by RP14 in tail sensory and motor neurons. 3. In contrast, bath application of SCPB increased spontaneous activity in RP14. Subsequent application of 5-HT to the bath, which still contained SCPB, inhibited RP14. Therefore, the effects of SCPB were essentially opposite to those of 5-HT on this inhibitory interneuron. 4. FMRFamide had little effect on RP14. It did not produce an obvious change in its resting membrane potential and produced only a transient increase in its spontaneous activity. 5. These results suggest that various neuromodulators have differential effects on elements of the neuronal circuit underlying the tail-withdrawal reflex of Aplysia. Differential modulation may determine the overall behavioral manifestations associated with sensitization.

Action Potentials↗

Identification, characterisation and in vitro reconstruction of an interneuronal network of the snail Helisoma trivolvis.

1. We describe three interneurones and their follower cells in the central ganglionic ring of Helisoma trivolvis. 2. The largest neurone on the dorsal surface of the left pedal ganglion is shown to be an interneurone that contains dopamine and makes monosynaptic connections with a large number of follower cells in the visceral and left parietal ganglia. This neurone is designated as left pedal dorsal 1 (LPeD1). 3. Another giant neurone is located on the dorsal surface of the right pedal ganglion. Although the position and morphology of this cell, designated right pedal dorsal 1 (RPeD1), are similar to those of LPeD1, it contains serotonin rather than dopamine. This neurone was found to synapse only on LPeD1, no other follower cells have so far been discovered. The connections between LPeD1 and RPeD1 are mutually inhibitory. 4. A small FMRFamide-immunoreactive neurone, identified here as visceral dorsal 4 (VD4), is located on the dorsal surface of the visceral ganglion. This neurone has a large number of follower cells throughout the central ganglionic ring. Among these follower cells are LPeD1 and RPeD1. The transmitter utilized by VD4 at these synapses is probably FMRFamide. In addition, VD4 receives excitatory inputs from LPeD1 that appear to be chemical and monosynaptic. 5. To test further the monosynaptic and specific nature of the connections within the network, the three interneurones were isolated and cultured in vitro. In these circumstances, the three neurones extended neurites and formed synapses which, with one exception (occasional electrical coupling between LPeD1 and RPeD1), were of similar type to those observed in vivo. 6. The identification and characterization of these three interneurones and their follower cells should greatly facilitate future studies of the Helisoma trivolvis nervous system. Furthermore, the possibility that this three-cell network can be reconstructed in vitro should aid our understanding of the mechanisms underlying synapse formation and neuronal plasticity.

Animals↗

Modulatory effects of some amino acids and neuropeptides on luminescence in the brittlestar amphipholis squamata

Amphipholis squamata is a polychromatic luminescent ophiuroid. The effects of amino acids ( &ggr; -aminobutyric acid, GABA, taurine, glycine and glutamate), N-methyl-d-aspartate (NMDA) and the invertebrate neuropeptides Antho-RFamide, FMRFamide and SALMFamides S1 and S2 were tested on acetylcholine-induced luminescence from isolated arms of clear and black specimens of Amphipholis squamata. The results showed that GABA, glycine and Antho-RFamide inhibited ACh-induced luminescence of clear specimens and had no significant effect on black specimens. Glutamic acid had no significant effect on ACh-induced luminescence, but triggered luminescence in the absence of ACh in both types of specimen. Taurine, NMDA and FMRFamide showed no significant effects on either clear or black specimens. S1 potentiated ACh-induced luminescence of clear and black specimens, while S2 had no clear modulatory effect on luminescence. These results suggest that, in addition to the previously described cholinergic system in Amphipholis squamata, there is also a modulatory component to luminescence control. Moreover, we observed a difference in modulation of luminescence between clear and black specimens.

Journal Article↗

Immunocytochemical studies on the islet and the gut of the arctic lamprey, Lampetra japonica.

The endocrine cells and nerves in the islet and the gut of the arctic lamprey Lampetra japonica were examined immunocytochemically by using antisera against brain-gut peptides and amine. The cellular composition of the islets as reported by previous researchers in European species of the lamprey was confirmed in the present study. The islet consisted exclusively of insulin immunoreactive cells in the larvae (ammocoetes), whereas in the adult somatostatin immunoreactive cells were added to the insulin immunoreactive cells; the gut epithelium in the adult was now devoid of somatostatin cells. In the gut of the lamprey, the endocrine cells--which were flask-shaped with a cytoplasmic process extended to the lumen--were classified into three types in the larvae, but were represented by a single type in the adult. In the larval lamprey, the first type was immunoreactive for somatostatin, the second one for gastrin/cholecystokinin (CCK) and the third cell type was immunoreactive for glucagon, pancreatic polypeptide and FMRFamide, simultaneously. In the gut of the adult lamprey, the single type of endocrine cell reacted simultaneously to C-terminal specific anti-glucagon serum, N-terminal specific anti-glucagon serum, anti-bovine PP serum, anti-neuropeptide Y serum and anti-FMRFamide serum. These cells occurred most frequently in the upper intestine, their distribution decreasing from the middle to the lower intestine. Two types of peptide containing nerves were identified in the islet and the gut of the larval and adult lamprey. The first type of neurons (perikarya and fibers) was immunoreactive for serotonin and calcitonin gene-related peptide (CGRP), and was located in the mucous and muscular layer of the intestine and in the islet. The second type of neurons contained both serotonin- and gastrin releasing peptide (GRP)-like immunoreactivities and was scattered exclusively in the muscular layer of the gut. In larval and adult lampreys, a few serotonin/CGRP immunoreactive nerve cell bodies and beaded fibers were found in the connective tissue around the islet cell cords. These nerve fibers were sometimes closely apposed to the blood capillaries and to the islet cells. These findings indicate that a neuroendocrine correlation comparable with that in mammals may have been established in the islet of this most primitive vertebrate.

Aging↗

An endogenous SCP-related peptide modulates ciliary beating in the gills of a venerid clam, Mercenaria mercenaria.

The activities of both the lateral and frontal cilia of Mercenaria mercenaria were unaffected, either by the two endogenous SCP-related peptides AMSFYFPRMamide and YFAFPRQamide, or by FMRFamide (all at 10(-6) M). Dopamine (DA) inhibited the lateral cilia; the mean EC50 was 2 x 10(-6) M. The peptide YFAFPRQamide--but neither AMSFYFPRMamide nor FMRFamide--antagonized the inhibition induced by DA; this effect was dependent on both time and dose. At a DA concentration of 5 x 10(-7) M, the effect of YFAFPRQamide appeared within 20 min and became maximal within 40-60 min; the mean EC50 at these times was 4.7 x 10(-11) M. If the concentration of DA was increased to 10(-6) M, the maximal effect of the peptide was delayed to 50 min, and the mean EC50 increased to 1.1 x 10(-7) M. Particle transport by the frontal cilia was inhibited by 5-hydroxytryptamine (5HT); the mean EC50 was 5.7 x 10(-7) M. Again, only YFAFPRQamide had an antagonistic effect on the 5HT-induced inhibition. At a 5HT concentration of 10(-6) M, the effects of YFAFPRQamide did not appear until 45 min; the mean EC50 was 10(-6) M. When radioimmunoassayed with an SCP antiserum, the elution profile of a gill extract overlapped those of the SCP-related peptides that had previously been identified in extracts of whole animals. These data suggest that all three SCP analogs occur in the gill. Immunohistochemistry of the gill, carried out with a monoclonal antibody raised to SCPB, stained many varicose neuronal fibers. Most of these were associated with the gill musculature, but a sparse innervation of the filaments underlying the cilia was also observed. Some fluorescent nerve cell bodies were also seen in the gill tissue. Our results are consistent with the hypothesis that YFAFPRQamide modulates branchial activities--muscular as well as ciliary--that are associated with feeding.

Animals↗

Interganglionic axonal transport of neuropeptides in Aplysia.

The transport of neuropeptides between central ganglia was studied in Aplysia. Peptide transport was determined by incubating ganglia with 35S-methionine and measuring the appearance of labeled peptides in connected ganglia. Selected interganglionic connectives were left intact and passed through a diffusion barrier separating the ganglia. Labeled peptides transported between ganglia included FMRFamide, myomodulin, and pedal peptide. Each of these peptides has been shown to be physiologically active in Aplysia. In addition to these previously characterized neuropeptides, a number of other as yet uncharacterized labeled peptides were also transported. All the peptides were transported by fast axonal transport as judged by the distance transported and/or the sensitivity to colchicine. Overall, FMRFamide and several unidentified peptides were the predominant transported peptides. However, the nature and amount of the peptides transported differed for each ganglia. These results support the proposition that the labeled peptides have transmitterlike actions and suggest that there are a number of neuropeptides that are likely to have central actions that have not yet been characterized in Aplysia.

Abdomen↗

Acetylcholine suppresses calcium current in neurons of Aplysia californica.

1. The left upper quadrant neurons L2-L6 in the abdominal ganglion of Aplysia californica were voltage clamped in order to examine effects of acetylcholine on voltage-dependent Ca and Ca-dependent K currents. 2. "Puffed" application of 10-100 microM acetylcholine reduced both the early inward and late outward phases of the current elicited by depolarizing voltage steps. An identical effect of the peptide FMRFamide was previously found to result from a suppression of the Ca and Ca-dependent K currents. 3. This effect of acetylcholine was obscured by the simultaneous activation of a previously described K current resembling the "S" current. Extracellular tetraethylammonium (TEA) and 4-aminopyridine could not be used to eliminate this current, because both compounds also appeared to block the acetylcholine receptor mediating the putative suppression of Ca and Ca-dependent K currents. 4. The acetylcholine-induced "S"-like and other K currents could, however, be reduced or eliminated by injection of TEA+ or Cs+ into the cell, replacement of extracellular Ca2+ with Ba2+, and by shifting the K+ equilibrium potential so as to null K currents at the potential used to record Ca current, revealing in each case a partial (10-40%) suppression of the Ca (or Ba) current by acetylcholine. 5. The reduction of the outward phase of depolarization-activated current was confirmed to represent suppression of the Ca-dependent K current by acetylcholine. This effect was indirect, secondary to the suppression of Ca current, since acetylcholine had no effect on Ca-dependent K current elicited by direct injection of Ca2+ into the cell. 6. Activation of the "S"-like K current and suppression of the Ca current by FMRFamide are likely to be important in its proposed role as an agent of presynaptic inhibition in Aplysia. Since acetylcholine has identical effects, it too may have such a function.

4-Aminopyridine↗

Recovery of tail-elicited siphon-withdrawal reflex following unilateral axonal injury is associated with ipsi- and contralateral changes in gene expression in Aplysia californica.

Behavioral, cellular and molecular changes were examined following axonal injury in the marine mollusc Aplysia californica. Unilateral nerve injury was performed by crushing the pleural-pedal connective and the peripheral pedal nerves innervating one side of the posterior body wall and the tail. The injury procedure severs the axons of the pleural sensory neurons resulting in the blockade of the tail-elicited siphon-withdrawal reflex. Partial reflex recovery is observed within 3 d and reaches 50% of the pretest value by six weeks postinjury. Retrograde staining of injured nerves combined with electrophysiological recordings from siphon motor neurons show that axons can regenerate through the crushed site and reconnect with the tail by three weeks postinjury. Moreover, the behavioral and electrophysiological measurements suggest that the contralateral sensory neurons may contribute to the early recovery of the siphon-withdrawal reflex. The levels of mRNAs coding for actin and calreticulin are elevated while the mRNAs coding for intermediate filament protein, sensorin A, FMRFamide are reduced in the ipsilateral pleural ganglia as detected by Northern blots. In the contralateral pleural ganglia, the levels of mRNAs coding for actin, sensorin A and FMRFamide are elevated. These molecular changes in both the ipsi- and contralateral sides are consistent with the hypothesis that both sides are participating in the behavioral recovery following unilateral axonal injury.

Actins↗

Neuromusculature of the ovijector of ascaris suum (Ascaroidea, nematoda): an ultrastructural and immunocytochemical study.

This study used electron microscopy and confocal scanning laser microscopy interfaced with cytochemistry to study neuromuscular interrelationships in the ovijector of Ascaris suum. An extensive nerve plexus with both FaRPergic and non-FaRPergic components extends over the outer surface of the ovijector. The non-FaRPergic component is derived from nerve branches of the ventral nerve cord, whereas the FaRPergic component emanates from two large FMRFamide-immunoreactive neurons. In the vagina vera, most myofibrils are circular in orientation and a number of them divide and run for short distances in longitudinal and diagonal directions, their myofilaments are also orientated in a variety of directions. Parallel nerve fibres run in tracts along the length of the vagina vera with branches that penetrate the muscle layers. The vagina uteri possesses a thicker hypodermis than that of the vagina vera. It appears rich in secretory and phagocytic vesicles and the luminal side is invested with an electron-dense substance. The musculature of the vagina uteri is less well developed than that of the vagina vera, being restricted to circular myofibrils, with an apparent diagonal arrangement of myofilaments. Also, the innervation is less extensive in the vagina uteri with many fibres returning to the vagina vera to rejoin the nerve net and others continuing into the uteri.

Animals↗

Cell type-specific regulatory sequences control expression of the Drosophila FMRF-NH2 neuropeptide gene.

The FMRFamide (dFMRFa) neuropeptide gene is expressed in about 17 diverse cell types in the Drosophila central nervous system. This expression pattern is generated by transcriptional control elements that are distributed over 8 kilobases of dFMRFa DNA. Previous studies identified one enhancer within the dFMRFa 5' region that is both necessary and sufficient to drive reporter transgene expression in one of the 17 dFMRFa cell types, the OL2 neurons. We now report the presence of two additional, non-overlapping enhancers within the gene: One drives expression by the six Tv neuroendocrine cells, and another in the four X and X2 interneurons. We also show that the Tv neuron-specific enhancer itself has complex organization, with several positively and negatively acting cis elements. Together, these results describe the organization of what is likely to be a prototypic neuronal gene promoter: an assemblage of multiple, independent, cell type-specific enhancers, each consisting of multiple quantitative elements.

Animals↗

Ultrastructural aspects of peptidergic modulation in the peripheral nervous system of Helix pomatia.

The ultrastructural characteristics of peptidergic peripheral contacts in the snail, Helix pomatia, were investigated, with special attention to the innervation of the heart, buccal mass, and salivary gland by Mytilus inhibitory peptide-immunoreactive neurons. Following the application of correlative light- and electron-microscopic pre-embedding immunocytochemistry, the peripheral tissues reveal a rich innervation by Mytilus inhibitory peptide-immunoreactive elements. These neurons establish three types of neuromuscular contacts in the heart and buccal mass: (1) close (16-20 nm) unspecialized membrane contacts; (2) contacts with a relative wide (40-100 nm) intersynaptic cleft; and (3) labeled varicosties located freely in the extracellular space, far (0. 5-several microm) from the muscle cells. In the salivary gland, the immunoractive profiles contact both the muscular and glandular elements with close (type 1) and wider (type 2) membrane attachments. The great majority of Mytilus inhibitory peptide-immunoreactive profiles contain an ultrastructurally uniform population of large (120-150 nm) electron dense granules. The ultrastructural features of the innervation by Mytilus inhibitory peptide-immunoreactive elements are compared with those established by immunogold labelled FMRFamide-containing profiles in the heart and salivary gland. These latter display similarities in forming the different kinds of intercellular contacts, and differences in the morphological variability of the content of granules in the immunolabeled profiles. The results suggest diverse, non-synaptic modulatory roles of neuropeptides in the peripheral nervous system of Helix pomatia, including localized membrane effects and neurohormonal-like remote global controls, that may also be of significance in orchestrating the effects of neuropeptides released at the same time on different targets.

Animals↗

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↗

Structure of the nervous system of Myzostoma cirriferum (Annelida) as revealed by immunohistochemistry and cLSM analyses.

The nervous systems of juvenile and adult Myzostoma cirriferum Leuckart, 1836, were stained with antisera against 5-HT (5-hydroxytryptamine, serotonin), FMRFamide, and acetylated alpha-tubulin in combination with the indirect fluorescence technique and analyzed by confocal laser scanning microscopy. The central nervous system consists of two small cerebral ganglia, connected by a dorsal commissure, a ventral nerve mass, and a pair of long circumesophageal connectives joining the former to the latter. The two neuropil cords within the ventral nerve mass curve outward and are joined to one another anteriorly and posteriorly. They are connected by 12 commissures, forming a ladder-like system. A single median nerve runs along the midventral axis. In addition to the circumesophageal connectives, 11 peripheral nerves arise from each main cord. The first innervates the anterior body region. The others form five groups of two nerves each, the first and thicker nerve of which is the parapodial nerve, innervating the parapodium and two corresponding cirri. Except for those in the most posterior group, the second nerves innervate the lateral organs and the body periphery. Serotonergic perikarya are arranged in six more or less distinct clusters, the first lying in front of and the other five between the main nerve cords. The distribution pattern of the FMRFamidergic perikarya is less clear and the somata lie between and outside the cords. One pair of dorsolateral longitudinal nerves was visualized by tubulin staining. Peripheral nerves and the commissures, in particular, demonstrate a segmental organization of the nervous system of M. cirriferum. Furthermore, their arrangement indicates that the body consists of six segments, the first of which is identifiable only by the first pair of peripheral nerves, the first two commissures, and the anteriormost ventral ganglion. The nervous system M. cirriferum thus exhibits several structures also found in the basic plan of the polychaete nervous system.

Animals↗

Neuronal development in larval chiton Ischnochiton hakodadensis (Mollusca: Polyplacophora).

Chitons are the most primitive molluscs and, thus, a matter of considerable interest for understanding both basic principles of molluscan neurogenesis and phylogeny. The development of the nervous system in trochophores of the chiton Ischnochiton hakodadensis from hatching to metamorphosis is described in detail by using confocal laser scanning microscopy and antibodies raised against serotonin, FMRFamide, and acetylated alpha tubulin. The earliest nervous elements detected were peripheral neurons located in the frontal hemisphere of posthatching trochophores and projecting into the apical organ. Among them, two pairs of unique large lateral cells appear to pioneer the pathways of developing adult nervous system. Chitons possess an apical organ that contains the largest number of neurons among all molluscan larvae investigated so far. Besides, many pretrochal neurons are situated outside the apical organ. The prototroch is not innervated by larval neurons. The first neurons of the developing adult central nervous system (CNS) appear later in the cerebral ganglion and pedal cords. None of the neurons of the larval nervous system are retained in the adult CNS. They cease to express their transmitter content and disintegrate after settlement. Although the adult CNS of chitons resembles that of polychaetes, their general scenario of neuronal development resembles that of advanced molluscs and differs from annelids. Thus, our data demonstrate the conservative pattern of molluscan neurogenesis and suggest independent origin of molluscan and annelid trochophores.

Animals↗

Molecular characterization and cell-specific expression of a Manduca sexta FLRFamide gene.

FMRFamide-related peptides (FaRPs) are a large group of neuropeptides containing a common RFamide C-terminus; they have been identified in vertebrates and invertebrates. We have isolated the cDNA that encodes three FaRPs in the tobacco hornworm, Manduca sexta, including the amidated decapeptide F10. The larger FaRPs are the partially processed precursors of F10, a neuropeptide belonging to the myosuppressin family of peptides. The presence of all three FaRPs in different tissues suggests differential utilization of typical dibasic processing sites and atypical processing sites C-terminal to leucine residues. F10 mRNA was detected in the brain, nerve cord, and midgut, and the mRNA levels in the nervous system are dynamically regulated during development. In situ hybridization analysis localized the F10 mRNA to a variety of cell types within the central nervous system (CNS), a peripheral neurosecretory cell (L1), and midgut endocrine cells, which suggests diverse functions. Distribution of the F10-containing neurons within the central nervous system is segment-specific, and the developmental profile suggests that the F10 gene products may have stage-specific functions. Molecular characterization of the F10 gene has provided insights into its regulation and cell-specific distribution that will enhance our understanding of how these FaRPs modulate different physiological systems and ultimately behavior.

Amino Acid Sequence↗

Neuronal development in larval polychaete Phyllodoce maculata (Phyllodocidae).

The existing view on neuronal development in polychaetes, as undergoing neurogenesis beginning in the rudiments of central ganglia and then extending peripherally, has been contrasted with the latest findings in molluscs, their sister trochozoan group, which show a peripheral to central mode of neurogenesis. The current study addresses this issue by examining early neuronal development in the polychaete Phyllodoce maculata using immunolabeling against acetylated alpha-tubulin, serotonin, and the FMRFamide. The first nervous cell was detected 20 hours before hatching, at the early trochophore stage. A solitary serotonergic neuron was located at the posterior-dorsal extreme of the larva and issued anterior projecting fibers, which outline the future ventral nerve cords and prototroch nerve. Two more serotonergic dorsal peripheral cells and three peripheral FMRFamidergic cells appeared soon thereafter. The fibers of these early cells formed a scaffolding, which prefigured the future adult nervous system (cerebral ganglion, ventral cords, prototroch and esophageal nerve rings) in prehatched trochophores. Shortly before hatching, the larval nervous system developed, including the apical organ, meridianal nerves in the episphere, and posttrochal nerves that innervate the feeding apparatus. After hatching, the rudiments of the adult nervous system started to develop along the paths already established by the earliest peripheral neurons. Thus, the general strategy of neurogenesis in a representative polychaete trochophore appears to resemble that of molluscs. The first neuronal cells to appear are peripheral in origin, located near the posterior margins of the embryo. Their similar anatomical appearance suggests that they share a similar functional role in trochophore development and behavior.

Animals↗