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PubMed2003

Organization and neural connections of the anterior optic tubercle in the brain of the locust, Schistocerca gregaria.

The anterior optic tubercle is a small neuropil in the insect brain and a major target of visual interneurons from the optic lobe. The functional role of the tubercle is poorly understood, but recent evidence from locusts points to a possible involvement in polarization vision. The present study examines the organization of the anterior optic tubercle in the locust Schistocerca gregaria and its connections with other brain areas. The tubercle of the locust consists of an upper and a lower subunit. Both units are connected in parallel with the medulla and lobula of the optic lobe, with the contralateral tubercle, and with the lateral accessory lobe in the median protocerebrum. Wide-field transmedullary neurons provide input from the medulla. Neurons with processes in the dorsal rim of the medulla, a relay station in the polarization vision pathway, project exclusively to the lower unit of the tubercle. Visual input from the lobula to the upper and lower unit originates from topographically distinct strata. The most prominent output target of the tubercle is the lateral accessory lobe in the median protocerebrum. Neurons from the upper unit project widely in the lateral accessory lobe, whereas neurons from the lower unit have focused projections confined to the median olive and to the lateral triangle. The two subunits of the anterior optic tubercle are, therefore, processing stages in two parallel visual pathways from the optic lobe to the median protocerebrum. Pathways via the lower unit of the tubercle appear to be involved in polarization vision.

Animals↗
PubMed2004

Centrifugal visual system of Crocodylus niloticus: a hodological, histochemical, and immunocytochemical study.

The retinopetal neurons of Crocodylus niloticus were visualized by retrograde transport of rhodamine beta-isothiocyanate or Fast Blue administered by intraocular injection. Approximately 6,000 in number, these neurons are distributed in seven regions extending from the mesencephalic tegmentum to the rostral rhombencephalon, approximately 70% being located contralaterally to the injected eye. None of the centrifugal neurons projects to both retinae. The retinopetal neurons are located in rostrocaudal sequence in seven regions: the formatio reticularis lateralis mesencephali, the substantia nigra, the griseum centralis tectalis, the nucleus subcoeruleus dorsalis, the nucleus isthmi parvocellularis, the locus coeruleus, and the commissura nervi trochlearis. The greatest number of cells (approximately 93%) is found in the nucleus subcoeruleus dorsalis. The majority are multipolar or bipolar in shape and resemble the ectopic centrifugal visual neurons of birds, although a small number of monopolar neurons resembling those of the avian isthmo-optic nucleus may also be observed. A few retinopetal neurons in the griseum centralis tectalis were tyrosine hydroxylase (TH) immunoreactive. Moreover, in the nuclei subcoeruleus dorsalis and isthmi parvocellularis, both ipsilaterally and contralaterally, approximately one retinopetal neuron in three (35%) was immunoreactive to nitric oxide synthase (NOS), and a slightly higher proportion (38%) of retinopetal neurons were immunoreactive for choline acetyltransferase (ChAT). Some of them contained colocalized ChAT and NOS/reduced nicotinamide adenine dinucleotide phosphate-diaphorase. Fibers immunoreactive to TH, serotonin (5-HT), neuropeptide Y (NPY), or Phe-Met-Arg-Phe-amide (FMRF-amide) were frequently observed to make intimate contact with rhodamine-labeled retinopetal neurons. These findings are discussed in relation to previous results obtained in other reptilian species and in birds.

Alligators and Crocodiles↗
PubMed2006

Mitral cells in the olfactory bulb of adult zebrafish (Danio rerio): morphology and distribution.

The mitral cell is the primary output neuron and central relay in the olfactory bulb of vertebrates. The morphology of these cells has been studied extensively in mammalian systems and to a lesser degree in teleosts. This study uses retrograde tract tracing and other techniques to characterize the morphology and distribution of mitral cells in the olfactory bulb of adult zebrafish, Danio rerio. These output neurons, located primarily in the glomerular layer and superficial internal cell layer, had variable-shaped somata that ranged in size from 4-18 microm in diameter and 31-96 microm2 in cross-sectional area. The mitral cells exhibited two main types of morphologies with regard to their dendrites: the unidendritic morphology was a single primary dendrite with one or more tufts, but multidendritic cells with several dendritic projections also were seen. The axons of these cells projected to either the medial or the lateral olfactory tract and, in general, the location of the cell on the medial or lateral side of the bulb was indicative of the tract to which it would project. Further, this study shows that the majority of zebrafish mitral cells likely innervate a single glomerulus rather than multiple glomeruli. This information is contrary to the multiple innervation pattern suggested for all teleost mitral cells. Our findings suggest that mitral cells in zebrafish may be more similar to mammalian mitral cells than previously believed, despite variation in size and structure. This information provides a revised anatomical framework for olfactory processing studies in this key model system.

Animals↗
PubMed1989

Immunohistochemical distribution and partial characterization of FLFQPQRFamidelike peptides in the central nervous system of rats.

Two peptides that are structurally related to the molluscan tetrapeptide Phe-Met-Arg-Phe-NH2 (FMRF-NH2) were recently isolated from bovine brain extract (Yang et al.: Proc. Natl. Acad. Sci. USA 82:7757-7761, '85). These peptides have an attenuating effect on morphine-induced analgesia when injected intracerebroventricularly in rats. Antisera against the two peptides--an octapeptide, Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH2 (F8F-NH2), and an octadecapeptide, Ala-Gly-Glu-Gly-Leu-Ser-Ser-Pro-Phe-Trp-Ser-Leu-Ala-Ala-Pro-Gln-Arg-Phe- NH2 (A18F-NH2)--were raised in rabbits and characterized with standard radioimmunoassay and immunohistochemical blocking controls. This study was aimed at localizing neurons in the rat brain that contain immunoreactivity for these peptides. Cryostat sections of normal and colchicine-treated Sprague-Dawley rat brains were incubated with the specific antisera and the immunoreactivity was visualized by the PAP or the FITC method. Immunoreactive neurons were detected in the hypothalamus and nucleus of the solitary tract. Relatively dense networks of fibers and terminals were observed in the lateral parabrachial nucleus and in the nucleus of the solitary tract. Fibers and terminals were also seen in the lateral septum, amygdala, hypothalamus, neurohypophysis, thalamus, periaqueductal gray, and several medullary nuclei. In these areas the immunoreactivity was abolished when the antisera were incubated with the corresponding synthetic peptides F8F-NH2 and A18F-NH2. Preincubation of the antisera with neuropeptide Y (NPY) did not affect the staining. The results suggest that there are neurons in the rat brain that contain F8F-NH2- and A18F-NH2-like peptides. The neuronal system described here may have a role in the regulation of autonomic functions, such as hypothalamic control and respiratory functions. The neuronal connections of the cells containing F8F-NH2- and A18F-NH2-like peptides remain to be studied.

Amino Acid Sequence↗
PubMed1990

Characterization and partial purification of different factors with contraction-potentiating activities from neurohaemal organs of the locust.

Phe-Met-Arg-Phe (FMRF-NH2) and structurally related peptides enhance neuromuscular transmission and contraction of the M. extensor tibiae preparations of the locusts Locusta migratoria and Schistocerca gregaria (Walther et al.: Neurosci. Lett. 45:99-104, '84). Similar effects could also be obtained with extracts of locust ganglia (Walther and Schiebe: Neurosci. Lett. 77:209-214, '87). By using two HPLC systems, we have partially purified extracts of the unpaired median nerves (including their neurohaemal organs) of different locust ganglia. The biological activity of the extracts served as an estimate for the degree of purification. Six different bioactive fractions were identified migrating at and close to retention times of known -RFamide peptides with similar bioactivity. No fraction coeluted with authentic FMRF-NH2 or FLRF-NH2. We demonstrate that extensor tibiae muscle contractions were potentiated by HPLC fractions from raw material with -RF-NH2 immunoreactivity, but also by HPLC fractions from raw material without such immunoreactivity.

Animals↗
PubMed1990

Co-localization of FLRF- and vasopressin-like immunoreactivity in a single pair of sexually dimorphic neurones in the nervous system of the locust.

The distribution of Phe-Leu-Arg-Phe (FLRF)-like immunoreactivity is described in the brain and in the ganglia of the ventral nerve cord of the locust Schistocerca gregaria. A single homologous pair of immunoreactive cell bodies occurs ventrally and medially in the suboesophageal ganglion. Each cell sends a process dorsally which bifurcates into anteriorly and posteriorly running neurites. The single anterior neurite passes along the circumoesophageal connectives to the brain where it ascends in a posterior running tract, giving off branches to innervate the tritocerebral neuropile and ending in an extensive network of highly varicose immunoreactive processes in the protocerebral neuropile. No processes are seen in the optic lobes or associated with the structured neuropiles of the muschroom bodies. The single posterior neurite from each cell passes into the suboesophageal-prothoracic connectives. It runs in the lateral dorsal tract of each ganglion in the ventral nerve cord as a highly varicose process and in each ganglion gives rise to an ipsilateral network of varicose processes in the dorsal and lateral neuropiles. In the seventh and terminal abdominal ganglia the innervation pattern exhibits sexual dimorphism. Vasopressin-like immunoreactivity is co-localized in the same pair of suboesophageal neurones and their processes. A similar pair of ventral median neurones stains with both antibodies in the suboesophageal ganglion of another species of locust, Locusta migratoria. Although the basic distribution pattern of immunoreactive processes is similar in both species there are also marked species differences in the pattern.

Amino Acid Sequence↗
PubMed1990

Organization of retinopetal axons in the optic nerve of the cichlid fish, Herotilapia multispinosa.

We have studied the position and numbers of retinopetal axons in the rainbow cichlid fish, Herotilapia multispinosa, to determine the response of related parts of the brain of fish to the continual addition of new neurons in the retina. The retinopetal axons were traced by using the retrograde tracers HRP and cobaltous lysine and an immunocytochemical probe, antibodies to FMRF amide, the molluscan cardioexcitatory peptide. One population of cells with retinopetal axons was found in the telencephalon (in the nucleus olfactoretinalis) and the other was scattered in the diencephalon. Some of the cells in the nucleus olfactoetinalis with retinopetal axons were FMRF amide positive; antibodies were used to trace the axons of these cells into the retina. All the retinopetal axons, from the nucleus olfactoretinalis and the diencephalon, were confined to the portion of the optic nerve that contains axons from the central retinal ganglion cells, that is, the oldest ganglion cells. This result suggests that the retinopetal axons grow into the optic nerve and retina early in the life of the fish, and no new ones are added later in life despite the extensive addition of cells in the retina. Counts of the cells in the nucleus olfactoretinalis that project to the retina in 3-month-old and adult fish support this interpretation. We conclude that retinopetal axons grow into the retina early in the life of the fish and respond to the formation of new retina by extending their arbors toward the new retina.

Animals↗
PubMed1994

Primary olfactory fibres project to the ventral telencephalon and preoptic region in trout (Salmo trutta): a developmental immunocytochemical study.

We studied the development of the primary olfactory system of a teleost, the brown trout, with the aims of clarifying whether the caudal projection pertains to the olfactory or to the terminal nerve system, of identifying the brain regions receiving this projection, and of investigating its possible functional significance. As olfactory markers (OMs) we used two polyclonal antibodies (to substance P and to alpha-melanocyte-stimulating hormone) that were found to label the olfactory projection strongly after preadsortion of the antibody with the corresponding antigen (OMs), and as a terminal nerve marker we used an antiserum to FMRF-amide peptide. OM labelling was observed in both perikarya and axons of olfactory neurons. In adults, olfactory neurons projected not only to olfactory glomeruli in the olfactory bulb but also, as has been reported previously, to more caudal targets in the forebrain through the medial olfactory tract. Our results show that these targets include the ventral and commissural nuclei of the area ventralis telencephali, the periventricular preoptic region, and the organum vasculosum laminae terminalis. Glomeruli were not observed before hatching, and the extrabulbar olfactory projections appear late in development. Extensive periventricular preoptic olfactory plexuses and olfactory innervation of the organum vasculosum laminae terminalis did not appear until adulthood. The cells of the ganglion nervus terminalis, which form ganglionic groups along the olfactory nerves, were not stained with these olfactory markers at any developmental stage studied, nor was the medial olfactory tract FMRP-amide peptide immunoreactive. Our results thus confirm the existence of primary olfactory projections to extrabulbar targets in trout. The target regions identified in this study are implicated in sexual behaviour: We discuss the related possibility that, in teleosts, these extrabulbar olfactory projections (rather than projections of the terminal nerve, as is widely held) are the primary mediators of neuroendocrine response to pheromones.

Animals↗
PubMed1995

Localization of the heptapeptide GFSKLYFamide in the sea cucumber Holothuria glaberrima (Echinodermata): a light and electron microscopic study.

Two peptides, Gly-Phe-Ser-Lys-Leu-Tyr-Phe-NH2 (GFSKLYFamide) and Ser-Gly-Tyr-Ser-Val-Leu-Tyr-Phe-NH2 (SGYSVLYFamide), recently isolated from the sea cucumber Holothuria glaberrima [Díaz-Miranda et al. (1992) Biol. Bull. 182:241-247] represent the first neuropeptides isolated from holothurians. Using an antibody against GFSKLYFa, we describe here the localization and distribution pattern of GFSKLYFa-like immunoreactivity in H. glaberrima, where immunoreactive fibers form a prominent and extensive peptidergic nervous system component. Neuron-like cells and nerve fibers expressing GFSKLYFa-like immunoreactivity are found in the ectoneural and hyponeural divisions of the radial nerve cords as well as in the digestive, haemal, respiratory, and reproductive systems; in the tentacles; and in tube feet. Neuroendocrine-like cells are found in the mucosal layer of the intestine. Ultrastructure immunocytochemical analysis revealed that, in nerve cells and fibers in the serosal layer of the intestine, the immunoreactivity is concentrated in vesicles. The immunoreactive nerve fibers are found mainly within a dense nerve plexus overlying and in close contact with smooth muscle cells of the intestine. The exclusive expression of GFSKLYFa-like immunoreactivity in neuronal or neuroendocrine tissue together with the close apposition of some fibers to muscle cells suggests that GFSKLYFa acts as a neuromuscular transmitter or neuromodulator in H. glaberrima. The wide occurrence of GFSKLYFa-like immunoreactivity throughout the nervous system of the sea cucumber suggests that GFSKLYFa plays an important role in the control of multiple action systems, including digestion, respiration, circulation, reproduction, and locomotion.

Animals↗
PubMed1994

Small cardioactive peptide B increases the responsiveness of the neural system underlying prey capture reactions in the pteropod mollusc, Clione limacina.

Effects of small cardioactive peptide B (SCPB) on cerebral neurons which underlie prey capture in the carnivorous pteropod mollusc, Clione limacina, were investigated. SCPB in concentrations of 10 microM and higher produced direct activation of cerebral ganglion neurons underlying extrusion of buccal cones used in prey capture. SCPB in lower concentrations, between 1 and 5 microM, did not have a noticeable effect on the membrane potentials of these neurons; however, it significantly increased their responsiveness to sensory inputs from the tactile stimulation of the head, and their ability to generate afterdischarge activity. SCPB immunoreactivity was observed in cell bodies in buccal, cerebral, pedal, and intestinal ganglia, as well as in the anterior esophagus and in buccal cones where fibers stained intensely. These electrophysiological and immunohistochemical data suggest that SCPB may have a physiological role in feeding arousal in Clione.

Action Potentials↗
PubMed2006

Neutral loss of water from the b ions with histidine at the C-terminus and formation of the c ions involving lysine side chains.

Neutral loss of water from the amide bond induced by the His side chain has been reported. The proposed fragmentation pathway is a retro-Ritter reaction catalyzed by the imidazole nitrogen. In our MS/MS study of the neuropeptide GAHKNYLRFamide, we observed that the neutral loss of water from the b(3) ion is abundant. The b(3) ion has a His residue at the C-terminus. As reported previously, in the b ions with His at the C-terminus, the imidazole residue is connected to the carbonyl carbon to form a five-membered ring. Therefore, it is unlikely that the neutral loss of water from the b(3) ion is catalyzed by the imidazole nitrogen. Through MS2 and MS3 studies of a synthetic peptide standard AGHKLL and its chemically labeled and isotope-encoded forms, we discovered that the water loss from the b(3) ion involves the carbonyl group of His, the hydrogen connected to the alpha-carbon of Gly, and the amide hydrogen of His. We also discovered the formation of an unusual c(x) ion in peptides with a Lys or Arg residue at the (x + 1) position of the peptide.

Acetylation↗
PubMed2005

Peptidomic analysis of the larval Drosophila melanogaster central nervous system by two-dimensional capillary liquid chromatography quadrupole time-of-flight mass spectrometry.

Peptides are the largest class of signalling molecules found in animals. Nevertheless, in most proteomic studies peptides are overlooked since they literally fall through the mazes of the net. In analogy with proteomics technology, where all proteins expressed in a cell or tissue are analyzed, the peptidomic approach aims at the simultaneous visualization and identification of the whole peptidome of a cell or tissue, i.e. all expressed peptides with their post-translational modifications. In this paper we describe the analysis of the larval fruit fly central nervous system using two-dimensional capillary liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (LC/Q-TOF-MS/MS. Using the central nervous systems of only 50 larval Drosophila as starting material, we identified 38 peptides in a single analysis, 20 of which were not detected in a previous study that reported on the one-dimensional capillary LC/MS/MS analysis of the same tissue. Among the 38 sequenced peptides, some originate from precursors, such as the tachykinin and the IFamide precursor that were entirely missed in the first study. This clearly demonstrates that the two-dimensional capillary LC approach enhances the coverage of the peptidomic analysis.

Amino Acid Sequence↗
PubMed1987

FMRF-amide-like immunoreactive efferent fibers and FMRF-amide suppression of pacemaker neurons in eyes of Bulla.

The eyes of certain marine gastropods including Aplysia and Bulla, contain circadian pacemakers, which produce a circadian rhythm of autogenous compound action potential (CAP) activity. The CAPs are generated by the synchronous spike discharge of a distinctive population of retinal pacemaker neurons whose axons convey the CAP activity to the CNS. When CAP activity is recorded from a preparation with eyes attached to the CNS, the CAP activity is modulated by efferent activity. In this study we have identified FMRF-amide-like immunoreactive efferent axons in the optic nerves of Bulla. These axons arborize in the basal retinal neuropil adjacent to the pacemaker neurons and are in a position to make synaptic contacts with their dendrites. Similar immunoreactive fibers are not observed in Aplysia eyes. Exogenous FMRF-amide at micromolar concentrations suppresses ongoing CAP activity in isolated eyes but does not suppress the ERG or phase shift the circadian rhythm of CAP activity. Intracellular recordings from the retinal pacemaker neurons reveal that FMRF-amide hyperpolarizes the membrane potential, suppresses spike discharge, and decreases the input resistance, suggesting that a K conductance is increased. Electrical stimulation of the region of the cerebral ganglion that contains FMRF-amide immunoreactive neurons suppresses ongoing CAP activity. All these results are consistent with the idea that the FMRF-amide immunoreactive central neurons and their axons provide a pathway for efferent modulation of the CAP rhythm generated by the retinal pacemaker neurons.

Animals↗
PubMed1998

Isolation and characterization of a novel bioactive peptide, Carassius RFamide (C-RFa), from the brain of the Japanese crucian carp.

A novel bioactive peptide with the C-terminal RFamide was isolated from the brain of the Japanese crucian carp, Carassius auratus langsdorfii, by using the intestine of the fish as the bioassay system. The primary structure of the peptide was determined to be SPEIDPFWYVGRGVRPIGRFamide and it was designated Carassius RFamide (C-RFa). The sequence of C-RFa was found to be significantly homologous to the molluscan neuropeptide termed Achatina cardioexcitatory peptide 1 (ACEP-1). Both peptides have RPXGRFamide structure in their C-terminal moieties. C-RFa was found to have an excitatory effect on visceral muscle tissues of fish, newt, quail, and rat. In the stomach of the fish, Zacco temminckii, IGRFamide (C-RFa 17-20) was shown to be the minimal structure required for the excitatory effect and PIGRFamide (C-RFa 16-20) to be almost equipotent with the parent peptide C-RFa.

Amino Acid Sequence↗
PubMed2000

Metamorphosis of tangential visual system neurons in Drosophila.

To learn about construction of the adult nervous system, we studied the differentiation of imaginal neurons in the Drosophila visual system. OL2-A and OL3 are tangential neurons that display dFMRFa neuropeptide gene expression in adults but not in larvae. The two large OL2-A neurons are generated near the end of the embryonic period and already show morphological differentiation at the start of metamorphosis. The numerous small OL3 neurons are generated postembryonically and first detected later in metamorphosis. The onset of dFMRFa transcription coincides with that of neuropeptide accumulation in OL2-A neurons, but it precedes peptide accumulation in the OL3 neurons by days. Altering each of the five conserved sequences within the minimal 256-bp OL dFMRFa enhancer affected in vivo OL transcriptional activity in two cases: alteration of a TAAT element greatly diminished and alteration of a 9-bp tandem repeat completely abolished OL2-A/OL3 reporter activity. A 46-bp concatamer containing the TAAT element, tested separately, was not active in OL neurons. We propose a model of neuronal differentiation at metamorphosis that features developmental differences between classes of imaginal neurons.

Amino Acid Sequence↗
PubMed1993

The distribution of immunoreactive FMRF-amide, neurotensin, and galanin in the brain and pituitary gland of three species of Xiphophorus from birth to sexual maturity.

This report seeks to extend the existing information on the relationship of regulatory neuropeptides to neuroendocrine and pituitary function through a longitudinal study of the distribution of FMRF-amide, galanin (GAL), and neurotensin (NT) in the brain and pituitary gland of three species of Xiphophorus from birth to sexual maturity. In the pituitary gland, immunoreactive (ir)-NT and -GAL were localized in the three regions of the adenohypophysis; ir-FMRF-amide was found in the neurohypophsis and in cells of the rostral pars distalis, but the immune reaction to this antisera in the pituitary was of very low intensity. Ir-GAL was found to colocalize with growth hormone, prolactin, and somatolactin in pituitary cells. In the brain, ir-GAL was found in the posterior nucleus lateralis tuberis (NLT), nucleus preopticus (NPO), and in the nucleus preopticus periventricularis (NPP). Ir-NT was localized in the anterior NLT. Ir-FMRF-amide was localized in the nucleus olfactoretinalis (NOR) where it was colocalized with gonadotropin releasing hormone, as well as in tracts that appeared to extend from the NOR, through the NPO and NPP, to the NLT. The NLT and the NOR have been previously implicated in the pituitary regulation of reproductive function. The above-cited localizations suggest that these peptides are involved in the neuroendocrine regulation of growth and reproduction in this genus.

Animals↗
PubMed1994

Endothelin-like immunoreactivity in midgut endocrine cells of the desert locust, Locusta migratoria.

Endothelin-1-like immunoreactivity has been found in endocrine cells of the midgut of the desert locust Locusta migratoria. Several antisera have been directed against the whole molecule and its C-terminal sequence. Endothelin-1-immunoreactive cells are present in the main region of the midgut (ventriculus) and in the midgut caeca but not in the ampullae through which the malpighian tubules drain. Endothelin-1-like immunoreactivity colocalizes with FMRFa immunoreactivity in the cells of the main region of the midgut but not in those in the midgut caeca. Endothelin-1-immunoreactive cells are present not only in adults but also throughout the five instars of posthatching development.

Animals↗
PubMed1982

Immunocytochemical demonstration of peptidergic cells in the pond snail Lymnaea stagnalis with an antiserum to the molluscan cardioactive tetrapeptide FMRF-amide.

With an antiserum to the molluscan cardioactive tetrapeptide FMRF-amide immunoreactive perikarya and nerve fibers were identified in the central and peripheral nervous system of the pond snail Lymnaea stagnalis. Their localization is described. The same antiserum yielded reactive product in particular cells of the epithelium of the alimentary tract. The use of two different fixatives, glutaraldehyde, and a mixture of glutaraldehyde, picric acid, and acetic acid (GPA) showed that certain nerve cells can be identified only in material fixed with either the one or the other of these two fixatives, a result which indicates that in Lymnaea more than one FMRF-amide-like substances may occur. "Positive" axon endings were found in the periphery of various nerves, i.e., in places where neurohormones are released into the blood. Other fibers were found to end, probably synaptically, on other neurons, on epithelial cells in the stomach, and between muscle cells in various parts of the body, e.g., in the heart. In these cases the FMRF-amide-like substance may function as a neurotransmitter or a neuromodulator.

Animals↗