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  • 1
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localisation and distribution of 5-hydroxytryptamine (5-HT, or serotonin) and a number of vertebrate neuropeptides in the nervous system of excysted (0–24 h) cysticercoid larvae of Hymenolepis diminuta were determined by an indirect immunofluorescence technique in wholemount preparations. In the central nervous system, cell bodies and nerve fibres immunoreactive to 5-HT are present in the main commissure, lateral and rostellar ganglia, and the longitudinal nerve cords and their connectives. In the peripheral nervous system, immunoreactive nerve fibres occur in a poorly developed nerve plexus within each sucker. Among the vertebrate peptides tested, antisera to pancreatic polypeptide (PP), polypeptide YY (PYY), peptide histidine isoleucine (PHI) and gastrin-releasing peptide (GRP) gave positive results. Immunoreactivity to PP and PYY paralleled that of 5-HT, with greater numbers of cell bodies present in the different locations within the scolex nervous system, and the sucker plexus being more prominent. The number of PP-reactive cells in the lateral ganglia and main lateral, longitudinal nerve cords increased over the 24-h period in culture. Results with antisera of different specificities to PP and PYY suggest that the immunoreactivity may be due to a peptide with closer structural affinity to PYY than to PP. Immunoreactivity to PHI is restricted to the main lateral nerve cords in the body of 0-h worms, extending into the median nerve cords by 12 h and 24 h. Immunoreactivity to GRP became evident after 12 h in culture and was confined to the longitudinal nerve cords, in particular the median nerve cords. The results are discussed in relation to the proposed transmitter and regulatory roles of 5-hydroxytryptamine and the neuropeptides.
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  • 2
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localisation and distribution of the cholinergic and serotoninergic components of the nervous system in the plerocercoid, adult and free proglottis stages of the tetraphyllidean tapewormTrilocularia acanthiaevulgaris were determined by enzyme histochemical and immunocytochemical techniques. The central nerve ring (CNR) in the scolex contains two lateral ganglia and gives rise to five pairs of longitudinal nerve cords (LNC's; three lateral, two median). The nerve cords run posteriorly throughout the bodies of the plerocercoid and adult worms and the free proglottis. Nerves from the CNR and accessory lateral LNC's pass to the bothridia, where they give rise to extensive nerve plexuses. As the individual proglottides develop along the strobila, a small nerve ring forms at the anterior end of each proglottis; within the nerve ring, distinct bilateral ganglia develop prior to the release of the proglottis. All ten LNC's are present in the free proglottis. The genital atrium and cirrus sac are innervated by cholinergic and serotoninergic elements. The cholinergic nervous system predominates in the CNS within the scolex, whereas there is a larger population of 5-HT-immunoreactive nerve cells associated with the LNC's and segmental ganglia along the strobila and within the free proglottis.
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  • 3
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization and distribution of seven neuropeptides in the nervous system of the plerocercoid, adult and free proglottis stages of the tetraphyllidean tapewormTrilocularia acanthiaevulgaris have been determined by an indirect immunofluorescence technique. Six of the peptides are vertebrate-derived, namely, pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), substance P (SP) and somatostatin (SRIF); the seventh is the invertebrate neuropeptide, FMRFamide. This is the first demonstration of VIP and SP immunoreactivity in a castode parasite, and for SRIF this is its first description in any parasitic platyhelminth. Cell bodies and nerve fibres immunoreactive to PP, PYY, VIP, SP and FMRFamide are present throughout the CNS; the distributions of PHI and SRIF were more restricted. In the PNS, nerve fibres immunoreactive to PP occur in the bothridia, whilst in the free proglottis nerve fibres immunoreactive to PYY and VIP innervate the gonads; VIP-immunoreactive nerve elements also supply the reproductive ducts. Extra-neuronal sitings of peptide immunoreactivities were evident for PHI, in association with the excretory system, and for SRIF, in presumed tegumental cell bodies in the free proglottis. The results are discussed in relation to the possible roles of the peptides in the neurophysiology and developmental biology of the worm.
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  • 4
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localistion and distribution of the cholinergic, serotoninergic and peptidergic components of the nervous system of the frog-lung flukeHaplometra cylindracea have been determined by the application of standard enzyme cytochemical and immunocytochemical techniques to cryostat sections and whole-mount preparations. Cholinesterase activity (ChE), as indicative of acetylcholine, has been demonstrated cytochemically in the CNS and PNS; however, the anterior ganglia were notably unreactive. The occurrence of serotonin was examined by an indirect immunofluorescence technique, and immunoreactivity (IR) was demonstrable in small, paired anterior ganglia and in fine nerve fibres associated with the somatic muscle, cirrus and gonopore. The peptidergic protion of the nervous system was investigated using antisera to 17 mammalian regulatory peptides and the invertebrate peptide FMRFamide, and was visualised by both indirect immunofluorescence and confocal scanning laser microscopy. Positive immunostaining occurred with antisera raised against pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), substance P (SP), peptide histidine isoleucine (PHI) and FMRFamide. Immunoreactivity to PP, PYY and FMRFamide was widespread throughout the nervous system and was evident in large, paired anterior ganglia, the dorsal commissure, main nerve tracts and the extensive array of small fibres that constitute the PNS. In contrast, the distribution of nerves immunoreactive to SP and PHI was less apparent, with PHI-IR occurring exclusively within the fibrous neuropile of the ganglia and in fibres of the ventral nerve cord. Results are discussed with respect to the distribution of the various neurochemical elements and their roles as putative neurotransmitters and/or regulatory molecules.
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  • 5
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localisation and distribution of neuropeptide F (NPF)-immunoreactivity (IR) in the monogenean fish-gill parasite,Diclidophora merlangi, have been investigated by whole-mount immunocytochemistry interfaced with confocal scanning laser microscopy and, at the ultrastructural level, by indirect immunogold labeling. Using antisera directed to intact synthetic NPF (Moniezia expansa, residues 1–39) or to the C-terminal decapeptide (residues 30–39) of synthetic NPF (M. expansa), immunostaining was found throughout the central (CNS) and peripheral nervous systems (PNS), including the innervation of the reproductive system. Immunoreactivity was found to be more intense using the antiserum to the C-terminal decapeptide fragment of NPF. At the subcellular level, gold labeling of NPF-IR was found exclusively over the contents of dense-cored vesicles that occupied nerve axons of both the CNS and the PNS. The distribution pattern of immunostaining for NPF mirrored exactly that previously documented for the vertebrate pancreatic polypeptide (PP) family of peptides and for FMRFamide. This finding and the results of preabsorption experiments strongly suggest that NPF is the predominant native neuropeptide inD. merlangi and that it accounts for most of the immunostaining previously obtained with PP and FMRFamide antisera.
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  • 6
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localisation and distribution of the cytoskeletal protein tubulin in the adult liver flukeFasciola hepatica have been determined by an indirect immunofluorescence technique using a monoclonal antibody raised against β-tubulin. Tubulin was demonstrated in the tegumental syncytium and in the tegumental cell bodies and their cytoplasmic connections with the surface syncytium. Immunostaining was also evident in the nerve fibres innervating sensory receptors in the tegument, in the nerve plexus innervating the sub-tegumental musculature and in the cytoplasmic extensions of the nurse cells within the vitelline follicle. Immunoblotting of whole fluke fractions produced a single band corresponding to a molecule of approximately 54 kDa in size. This figure corresponds with previous data obtained on tubulin from other helminth and eukaryotic sources.
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  • 7
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Both whole-mount preparations and cryostat sections of the monogenean, Diclidophora merlangi, were screened for the presence of 5-hydroxytryptamine-like immunoreactivity, using an indirect immunofluorescence technique. Immunostaining was evident in cell bodies and fibres comprising the longitudinal cords of the central nervous system and in the peripheral plexuses that innervate the musculature of the adhesive organs, feeding apparatus and accessory ducts of the reproductive system. The subsurface nerve-net was reactive, and a distinctive “collar” of staining marked a plexus of nerves and somata that supply the subtegumental muscles of the extensile forebody.
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  • 8
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localisation and distribution of 5-hydroxytryptamine (5-HT, or serotonin) and neuropeptides in the nervous system of the protoscolex of the hydatid organismEchinococcus granulosus were determined by an indirect immunofluorescence technique. Nerve-cell bodies immunoreactive for 5-HT occurred in the lateral ganglia and in association with the lateral longitudinal nerve cords. 5-HT immunostaining was also evident in the central nerve ring, in the rostellar nerves and in the nerve plexus innervating the suckers. Of the antisera used to screen the protoscolex for neuropeptide immunoreactivity (IR), immunostaining was obtained with those raised against pancreatic polypeptide (PP), peptide YY (PYY), substance P (SP), peptide histidine isoleucine (PHI) and vasoactive intestinal peptide (VIP). The most extensive pattern of IR occurred with antisera to PP and PYY. Immunoreactive nerve elements were evident in the lateral ganglia, central nerve ring, rostellar nerves, rostellar ganglia, sucker plexus and longitudinal nerve cords. The distribution of SP-, PHI- and VIP-IRs was more restricted: SP-IR occurred in the lateral ganglia and sucker nerves, whilst PHI- and VIP-immunoreactive nerve elements were associated with the lateral longitudinal nerve cords. Protoscoleces cultured in vitro for 29 days were also examined and neuroanatomical changes noted. A greater development of the longitudinal nerve cords and their cross-connectives in the body of the worm was evident, and a group of nerve cells were seen to develop at the posterior end of the main lateral nerve cords.
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  • 9
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Whole mounts of the metacercariae ofDiplostomum sp. andCotylurus erraticus from rainbow trout have been treated cytochemically for the demonstration of cholinergic, serotoninergic (5-hydroxytryptamine) and peptidergic elements in the nervous system. Antisera directed against four vertebrate (pancreatic polypeptide, peptide YY, substance P and peptide histidine isoleucine) and two invertebrate peptides (neuropeptide F and FMR Famide) were used in an indirect immunofluorescence procedure in conjunction with confocal scanning laser microscopy (CSLM). Of the seven antisera tested, all except peptide histidine isoleucine showed significant immunoreactivity. Cholinergic and serotoninergic staining was found primarily in the central nervous system (CNS) and in cell bodies associated with the ventral and dorsal nerve cords in both trematodes. Peptidergic immunoreactivity was localised in the CNS and PNS of both genera, revealing an extensive innervation within the holdfast organ and in and around the oral and ventral suckers.
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  • 10
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The nature of xenopsin immunoreactivity in mammalian antral G-cells has been reassessed. Xenopsin immunostaining was most intense in human antral G-cells, present in those of the dog and pig and not detected in guinea pig or rat tissues. Rigorous specificity controls for ionic binding of immunoglobulins to antral G-cell granules indicated that this mechanism was not responsible for xenopsin immunostaining. Preincubation of the xenopsin antiserum with xenopsin, human gastrin 1–13 and gastrin 2–17 completely abolished immunostaining at similar molar concentrations. Gastrin 34 was ineffective at much higher concentrations. These results infer that xenopsin-immunoreactivity in antral G-cells resides in the N-terminal region of gastrin 17. Examination of the primary structures of xenopsin and the N-terminal regions of some mammalian gastrins reveals a hitherto unrecognized homology.
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