Monoamine- and peptide-containing elements in the nemertine digestive tract

Detailed comparative-histological data on localization and morphological peculiarities of catecholamine-, serotonin-, neurotensin-and FMRFamide-containing elements in the nemertine digestive system are presented for the first time. Using fluorescent histochemistry and immunohistochemistry, pharynx, esophagus, and midgut were studied in five species from three genera of the White Sea nemertines. Described in various nemertine species are intra-and subepithelial cells of the open and closed type, containing biologically active substances. Their processes are distributed in basi-and subepithelial nerve plexuses and can make contact with lumen of the digestive tract. Species-specificity in localization of the cells containing certain biologically active substances is noticed both along the length of the tract and with respect to its epithelial layer. Peculiarities and common regularities in distribution are discussed, as well as a possible function of monoamine-and peptide-containing elements in the digestive tract of the studied nemertines and other invertebrates.     

Localization and distribution of catecholaminergic structures in the nervous system of Phocanema decipiens (nematoda)

The nervous system of Phocanema decipiens was examined with both the formaldeyhyde-induced and the glyoxylic acid fluorescence histochemical techniques. Green catecholaminergic structures were observed in 4 cephalic papillary nerves, 2 fibres with varicosities in the nerve ring as well as the ventral nerve cord and a pair of lateral nerves.The papillary nerves, extending from the nerve ring to the lips region, have cell bodies which are located anterior or adjacent to the nerve ring. Cell bodies of the lateral nerves are found within the lateral cord tissue posterior to the nerve ring. Each of these neurons has 3 processes—one joins with the nerve ring, the other merges with the ventral nerve cord and the third ends abruptly within the lateral cord.     

Monoamine- and peptide-containing elements in the nemertine digestive tract

Detailed comparative-histological data on localization and morphological peculiarities of catecholamine-, serotonin-, neurotensin- and FMRFamide-containing elements in the nemertine digestive system are presented for the first time. Using fluorescent histochemistry and immunohistochemistry, pharynx, esophagus, and midgut were studied in five species from three genera of the White Sea nemertines. Described in various nemertine species are intra- and subepithelial cells of the open and closed type, containing biologically active substances. Their processes are distributed in basi- and subepithelial nerve plexuses and can make contact with lumen of the digestive tract. Species-specificity in localization of the cells containing certain biologically active substances is noticed both along the length of the tract and with respect to its epithelial layer. Peculiarities and common regularities in distribution are discussed, as well as a possible function of monoamine- and peptide-containing elements in the digestive tract of the studied nemertines and other invertebrates.     

Presence and distribution of immunoreactive and bioactive FMRFamide-like peptides in the nervous system of the horseshoe crab, Limulus polyphemus

FMRFamide immunoreactivity was detected in all regions of the Limulus nervous system, including the brain (6.5 +/- 0.6 pg FMRFamide/mg), cardiac ganglion (2.06 +/- 0.67 pg FMRFamide/mg), and ventral nerve cord (5.8 +/- 0.7 pg FMRFamide/mg). The distribution of immunoreactive FMRFamide (irFMRFamide) was mapped by immunofluorescence and the distribution corresponded to regional RIA data. A good proportion of the CNS and cardiac ganglion neuropile contained irFMRFamide, and fluorescent cell bodies were observed in several areas. High performance liquid chromatography (HPLC) was employed to separate and characterize the FMRFamide-like peptides from extracts of Limulus brains. HPLC fractions were analyzed using coincidental radioimmunoassay and bioassay (the radula protractor muscle of Busycon contrarium). There appear to be at least three FMRFamide-like peptides in the Limulus brain, including one similar to clam FMRFamide. FMRFamide acts on Limulus heart in a biphasic manner at relatively high concentrations (10(-5)M), but has no effect on the activity of the isolated ventral nerve cord. These data suggest that in Limulus FMRFamide-like peptides are acting as neurotransmitters, or neuromodulators.     

Characterization and distribution of FMRFamide immunoreactivity in the rat central nervous system

FMRFamide immunoreactive material (irFMRFamide) was studied in rat brain and gastrointestinal tract. Highest irFMRFamide concentrations were found in tissues of the gastrointestinal tract and, in the brain, highest concentrations were found in the hippocampus, midbrain, brainstem and hypothalamus. High pressure liquid chromatographic characterization of irFMRFamide demonstrated that the immunoreactive material in brain, pancreas and duodenum was different from molluscan FMRFamide but it was also distinct from any known neuropeptide.     

Ultrastructural and immunocytochemical observations of the nervous systems of three macrodasyidan gastrotrichs

The nervous systems of three macrodasyidan gastrotrichs, Dactylopodola baltica, Macrodasys caudatus and Dolichodasys elongatus, were investigated using immunocytochemistry and electron microscopy. Labelling of neural structures against serotonin revealed the presence of two pairs of cerebral cells, a dorsal cerebral connective, and paired ventral nerve cords in D. baltica. In M. caudatus and D. elongatus serotonin immunoreactivity was present in a single pair of dorsal cerebral cells and the ventral nerve cords; the dorsal connective of D. elongatus was also immunoreactive to serotonin and acetylated α‐tubulin. The presence of paired, serotonin‐like immunoreactive cells in D. baltica and other species may represent the plesiomorphic condition in Macrodasyida. The fine structure of the photoreceptors in D. baltica was also investigated to explore the potential ground pattern for eyes in the Macrodasyida. The pigmented photoreceptors of D. baltica contain a unicellular pigment cup, sheath cell and sensory receptor. The pigment cup contains numerous osmiophilic granules that presumably function to shield the eyes from downwelling light in the red part of the spectrum. Projecting into the pigment cup and sheath cell are numerous microvilli from a bipolar sensory cell. A single sensory cell may represent the plesiomorphic condition in Macrodasyida, with multiplication of sensory cells representative of more derived taxa.     

柞蚕幼虫期神经系统一氧化氮合酶的NADPH-d组织化学

运用NADPH-d组织化学整体染色方法研究柞蚕Antheraea pernyi Gu rin-Meneville幼虫神经系统中一氧化氮合酶阳性细胞的分布、数量及形态特征。结果表明,柞蚕各龄期幼虫中枢神经的脑及各神经节中都有一氧化氮合酶阳性反应,阳性神经元根据其形态大小和染色特性可分为A,B,C3种类型:A型细胞沿神经节中线分布,阳性反应较强,胞体长径约30~70μm,各神经节中的数量恒定。B型细胞多分布于神经节周边部分,阳性反应较弱,胞体长径约8~20μm,各神经节中的数量变化较大,随着龄期增加有减少的趋势。C型细胞分布于咽下神经节和第8腹神经节,在3种细胞中阳性反应最强,胞体长径约16~50μm。     

Biogeographic patterns and the evolution of eureptantic nemerteans

The origin and evolution of the eureptantic nemerteans is discussed from a biogeographic point of view. It is most likely that East Indian Ocean was part of the ancestral distribution of the Eureptantia. The area cladogram estimated by Brooks parsimony analysis (BPA) is to a high degree congruent with a vicariance explanation of the evolution of the Eureptantia and suggests an ancestral distribution concordant with the Tethys Sea. A general area cladogram based on a combined BPA analysis of eureptantic nemerteans and acanthuroid fishes is reconstructed and suggested as a hypothesis of the relationships between east Indian Ocean, west Indian Ocean, west Pacific Ocean, east Adantic Ocean, west Atlantic Ocean, and the Mediterranean. This tree is compared with cladograms from the same areas based on other taxa.     

Phylogeny and cladistic classification of terrestrial nemerteans: the genera Pantinonemertes Moore & Gibson and Geonemertes Semper

Evolutionary relationships among seven species from two genera ( Pantinonemertes and Geonemertes ) of terrestrial nemerteans (phylum Nemertea) were estimated by a Wagner analysis of 27 morphological, ecological and life history characters. Pantinonemertes was found to be a paraphyletic taxon, and it is suggested that this genus should be combined with Geonemertes to form a monophyletic group. A previous hypothesis for how terrestrial nemerteans evolved is discussed and compared to the results of the cladistic analysis. This suggests that terrestrial and semi-terrestrial species evolved from a marine ancestor; the upper littoral and marine environments have been recolonized by descendants from a terrestrial ancestor.     

Expression of a novel neuropeptide, NVGTLARDFQLPIPNamide, in the larval and adult brain of Drosophila melanogaster

Advances in mass spectrometry and the availability of genomic databases made it possible to determine the peptidome or peptide content of a specific tissue. Peptidomics by nanoflow capillary liquid chromatography tandem mass spectrometry of an extract of 50 larval Drosophila brains, yielded 28 neuropeptides. Eight were entirely novel and encoded by five not yet annotated genes; only two genes had a homologue in the Anopheles gambiae genome. Seven of the eight peptides did not show relevant sequence homology to any known peptide. Therefore, no evidence towards the physiological role of these 'orphan' peptides was available. We identified one of the eight peptides, IPNamide, in an extract of the Drosophila adult brain as well. Next, specific antisera were raised to reveal the distribution pattern of IPNamide and other peptides from the same precursor, in larval and adult brains by means of whole-mount immunocytochemistry and confocal microscopy. IPNamide immunoreactivity is abundantly present in both stages and a striking similarity was found between the distribution patterns of IPNamide and TPAEDFMRFamide, a member of the FMRFamide peptide family. Based on this distribution pattern, IPNamide might be involved in phototransduction, in processing sensory stimuli, as well as in controlling the activity of the oesophagus.     

Distribution of NADPH-diaphorase activity in the central nervous system of the young and adult land snail Megalobulimus abbreviatus

Nitric oxide (NO) is a gas produced through the action of nitric oxide synthase that acts as a neurotransmitter in the central nervous system (CNS) of adult gastropod mollusks. There are no known reports of the presence of NOS-containing neurons and glial cells in young and adult Megalobulimus abbreviatus. Therefore, NADPH-d histochemistry was employed to map the nitrergic distribution in the CNS of young and adult snails in an attempt to identify any transient enzymatic activity in the developing CNS. Reaction was observed in neurons and fibers in all CNS ganglia of both age groups, but in the pedal and cerebral ganglia, positive neurons were more intense than in other ganglia, forming clusters symmetrically located in both paired ganglia. However, neuronal NADPH-d activity in the mesocerebrum and pleural ganglia decreased from young to adult animals. In both age groups, positive glial cells were located beneath the ganglionic capsule, forming a network and surrounding the neuronal somata. The trophospongium of large and giant neurons was only visualized in young animals. Our results indicate the presence of a nitrergic signaling system in young and adult M. abbreviatus, and the probable involvement of glial cells in NO production.     

Serotonergic and FMRFamidergic nervous systems in gymnolaemate bryozoan larvae

A growing body of data from nervous systems of marine invertebrate larvae provides an ideal background for comparisons among higher taxa. The currently available data from Bryozoa, however, do not allow for a consistent hypothesis of an ancestral state for this taxon, which would be necessary for phylogenetic inferences. The larval nervous systems of the four gymnolaemate species Flustrellidra hispida, Bugula fulva, Alcyonidium gelatinosum, and Bowerbankia gracilis are examined by means of antibody staining against the neurotransmitters serotonin and FMRFamide, as well as against acetylated α-tubulin. Despite considerable variation, a comparison reveals a common pattern of the distribution of serotonin. The neurotransmitter is found in at least two cells in the apical organ as well as in paired axial and lateral nerves emerging from a central nerve nodule. A ring nerve is present below the corona and at least two serotonergic cells are found between the corona cells. Serotonergic coronal cells might represent unique bryozoan features, whereas the remaining elements show resemblance to the situation found in most spiralian taxa. The data do not provide support for a closer relationship of Bryozoa to Phoronida or Brachiopoda.     

The nervous system of Tricladida. I. Neuroanatomy ofProcerodes littoralis (Maricola,Procerodidae): An immunocytochemical study

The organization of the nervous system ofProcerodes littoralis (Tricladida, Maricola, Procerodidae) was studied by immunocytochemistry, using antibodies to authentic flatworm neuropeptide F (NPF) (Moniezia expansa). Compared to earlier investigations of the neuroanatomy of tricladid flatworms, the pattern of NPF immunoreactivity inProcerodes littoralis reveals differences in the following respects: 1. Shape and structure of the brain. 2. Number and composition of longitudinal nerve cords. 3. Shape of branches of, and transverse connections between, main ventral nerve cords. 4. Composition of the pharyngeal nervous system. The rich innervation by NPF immunoreactive (IR) fibres and cells of the subepithelial muscle layer, the pharynx musculature and the musculature of the male copulatory apparatus indicates a neurotransmitter or neuromodulatory influence on muscular activity.     

Morphology and systematics in mesopsammic nemerteans of the genus Ototyphlonemertes (Nemertea,Hoplonemertea, Ototyphlonemertidae)

Interstitial nemerteans of the genus Ototyphlonemertes are difficult to organize into traditional morphospecies. They occur in a multitude of slightly different local varieties that form a seemingly continuous morphological cline. In this paper, we summarize most published morphological data on the group, plus 73 new records of geographic varieties from the Mediterranean Sea in the East to the Sea of Japan in the West. We summarize morphological variation, partition traits into character and character states, propose a standardized protocol for examination of live specimens and discuss the phylogenetic structure of the group. The phylogenetic discussion leads to a hypothesis that partitions all varieties (including the established species) into six groups. These are the smallest morphologically homogenous sets of varieties (corresponding to traditional morphospecies) we can diagnose on phylogenetically reliable traits. Variation within the groups appears to be unreliable phylogenetic markers that may distinguish ecological forms rather than relatedness. However, we distinguish four of the groups by combinations of two traits, one apomorphy for a more inclusive group and one plesiomorphy, and the remaining two by one trait each that may be either a unique plesiomorphy or an apomorphy depending on the rooting, and most of them may thus join paraphyletic groups of cryptic monophyletic units. We call this kind of group phylomorph and name them the Duplex-, Pallida-, Cirrula-, Fila-, Lactea- and Macintoshi-morph (referring to the first established species within each group, i.e. Ototyphlonemertes duplex, O. pallida, O. cirrula, O. fila, O. lactea and O. macintoshi respectively). The phylogenetic scheme provides a simple tool to allocate geographical varieties to a group of possible `species' and a phylogenetic null-hypothesis for further tests with genetic data.     

Distribution of peptide-containing neurons and endocrine cells in the rabbit gastrointestinal tract,with particular reference to the mucosa

Summary The distribution patterns of peptide-containing neurons and endocrine cells were mapped in sections of oesophagus, stomach, small intestine and large intestine of the rabbit, by use of standard immunohistochemical techniques. Whole mounts of separated layers of ileum were similarly examined. Antibodies raised against vasoactive intestinal peptide (VIP), substance P (SP), somatostatin (SOM), neuropeptide Y (NPY), enkephalins (ENK) and gastrin-releasing peptide (GRP) were used, and for each of these antisera distinct populations of immunoreactive (IR) nerve fibres were observed. Endocrine cells were labelled by the SP, SOM or NPY antisera in some regions.VIP-IR nerve fibres were common in each layer throughout the gastrointestinal tract. With the exception of the oesophagus, GRP-IR nerve fibres also occurred in each layer of the gastrointestinal tract; they formed a particularly rich network in the mucosa of the stomach and small intestine. Fewer nerve fibres containing NPY-IR or SOM-IR were seen in all areas. SOM-IR nerve fibres were very scarce in the circular and longitudinal muscle layers of each area and were absent from the gastric mucosa. The SP-IR innervation of the external musculature and ganglionated plexuses in most regions was rather extensive, whereas the mucosa was only very sparsely innervated. ENK-IR nerve fibres were extremely rare or absent from the mucosa of all areas, although immunoreactive nerve fibres were found in other layers.These studies illustrate the differences in distribution patterns of peptide-containing nerve fibres and endocrine cells along the gastrointestinal tract of the rabbit and also show that there are some marked differences in these patterns, in comparison with other mammalian species.     

An FMRFamide antiserum differentiates between populations of antigens in the ventral nervous system of the locust,Schistocerca gregaria

Summary The distribution of FMRFamide immunoreactive neurones in the ventral nerve cord of the locust, Schistocerca gregaria, is described. These neurones are found only in the suboesophagael and thoracic ganglia, although immunoreactive processes are found in the neuropils of the abdominal ganglia. Many of these neurones also react with an antiserum raised against bovine pancreatic polypeptide (BPP), but this antiserum also reveals another population of cells in the abdominal ganglia. The staining obtained with the BPP antiserum is blocked by preabsorption of the antiserum with FMRFamide; the converse is not true: FMRFamide-like immunoreactivity is not suppressed by preincubation with BPP. These results suggest that there are at least two endogenous peptide antigens in the locust nerve cord: one is found in cells of the suboesophageal and thoracic ganglia, and the other is found in cells of the abdominal ganglia.     

Acetylcholinesterase activity in the developing and regenerating nervous system of the acoel Symsagittifera roscoffensis

Bery, A. and Martínez, P. 2010. Acetylcholinesterase activity in the developing and regenerating nervous system of the acoel Symsagittifera roscoffensis. —Acta Zoologica (Stockholm) 92 : 383–392. The use of the cholinergic system is widespread in the animal kingdom. It controls different processes, including reproduction and neural transmission. However, its evolutionary history is not yet well understood. For instance, the role played by the cholinergic system in the nervous system of basal bilaterian taxa, where the first signs of architectural complexity appear, is still unknown. Here, we describe the structure of the cholinergic system during the development and regeneration of the acoel flatworm Symsagittifera roscoffensis, using acetylcholinesterase (AchE) activity as a marker. In this species, AchE activity is observed at all developmental stages, including in the early embryos. The juvenile and adult patterns reveal the presence of a complex nervous system that includes three pairs of longitudinal neurite bundles, which are connected to an anterior centralized mass of neurons and neural processes formed by two pairs of connectives and four commissures. The power of the technique also allows the detection of newly born neurons as they are incorporated into the growing nervous system (during regeneration).     

Effects of betel chewing on the central and autonomic nervous systems

Betel chewing has been claimed to produce a sense of well-being, euphoria, heightened alertness, sweating, salivation, a hot sensation in the body and increased capacity to work. Betel chewing also leads to habituation, addiction and withdrawal. However, the mechanisms underlying these effects remain poorly understood. Arecoline, the major alkaloid of Areca nut, has been extensively studied, and several effects of betel chewing are thought to be related to the actions of this parasympathomimetic constituent. However, betel chewing may produce complex reactions and interactions. In the presence of lime, arecoline and guvacoline in Areca nut are hydrolyzed into arecaidine and guvacine, respectively, which are strong inhibitors of GABA uptake. Piper betle flower or leaf contains aromatic phenolic compounds which have been found to stimulate the release of catecholamines in vitro. Thus, betel chewing may affect parasympathetic, GABAnergic and sympathetic functions. Betel chewing produces an increase in heart rate, blood pressure, sweating and body temperature. In addition, EEG shows widespread cortical desynchronization indicating a state of arousal. In autonomic function tests, both the sympathetic skin response and RR interval variation are affected. Betel chewing also increases plasma concentrations of norepinephrine and epinephrine. These results suggest that betel chewing mainly affects the central and autonomic nervous systems. Future studies should investigate both the acute and chronic effects of betel chewing. Such studies may further elucidate the psychoactive mechanisms responsible for the undiminished popularity of betel chewing since antiquity.