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Biomedical subjects

D W Halton

Publications and source records attributed to D W Halton.

At least 91 records · Page 5Linked to original sources
PubMed1994

Platyhelminth FMRFamide-related peptides (FaRPs) contract Schistosoma mansoni (Trematoda: Digenea) muscle fibres in vitro.

Molluscan FMRFamide and two recently discovered platyhelminth FMRFamide-related peptides (FaRPs), GNFFRFamide from the cestode Moniezia expansa and RYIRFamide from the terrestrial turbellarian Artioposthia triangulata, cause dose-dependent contractions of individual muscle fibres from Schistosoma mansoni in vitro. The most potent FaRP tested was the turbellarian peptide RYIRFamide, which produced a concentration-dependent effect between 10(-9) and 10(-7) M. FMRFamide and GNFFRFamide were less potent, inducing contractions between 10(-8)-10(-6) M and 10(-7)-10(-5) M respectively. The contractile effect of each of these peptides was blocked by the presence of 1 microM FMR-D-Famide. FMRF free acid did not elicit contraction of the muscle fibres. The FaRP-induced contractions did not occur if the Ca2+ was omitted and 0.5 microM EGTA was added to the extracellular medium. The FaRP-induced contractions were not blocked by the Ca2+ channel blockers nicardipine, verapamil or diltiazem, although high K+-induced contractions of these fibres were blocked by nicardipine. These data indicate the presence of FaRP receptors on schistosome muscle fibres and demonstrate their ability to mediate muscle contraction. The action of these endogenous flatworm peptides on schistosome muscle is the first demonstration of a direct excitatory effect of any putative neurotransmitter on the muscle of a flatworm, and establishes a role for FaRPs in neuromuscular transmission in trematodes. In addition, it provides the first evidence that the peptidergic nervous system is a rational target for chemotherapeutic attack in parasitic platyhelmiths.

Amino Acid Sequence↗
PubMed1994

A comparative study of the utilization of ether- and ester-linked phospholipid-containing liposomes by J774.E1 macrophage cell-line infected with Leishmania mexicana mexicana amastigotes.

The macrophage cell-line J774.E1 and Leishmania m. mexicana infection was used to investigate the uptake of liposomes, which differed in their bulk phospholipid: ester- or ether-analogue of phosphatydilcholine (PC). The receptor-mediated uptake of both species of liposomes, containing native or acetylated LDL as ligands was also evaluated. Uninfected and infected J774.E1 cell-line accumulated more ester- and ether-liposomes alone than mixed type (50:50, ester/ether). The utilization was significantly enhanced when both types of liposomes contained native LDL. The highest uptake was recorded for liposomes bearing acetylated LDL by infected J774.E1 cells. Accumulation of ester- and ether-liposomes with the same ligand was not markedly affected by different chemical nature of PC. Finally, ether-liposomes alone possessed certain activity against Leishmania m. mexicana amastigotes. The results presented here demonstrated the usefulness of ether-liposomes with specific ligands in site-specific delivery of antileishmanial compounds in vitro.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1994

5-Hydroxytryptamine (serotonin)-immunoreactivity in the nervous system of Mesocestoides corti tetrathyridia (Cestoda: Cyclophyllidea).

An indirect immunocytochemical technique has been interfaced with confocal scanning laser microscopy to investigate the occurrence and distribution of serotoninergic (5-HT) nerve elements in Mesocestoides corti tetrathyridia. Cell bodies and nerve fibers immunoreactive to 5-HT were found concentrated in the innervation around the 4 suckers and associated commissures and in the 5 pairs of longitudinal nerve cords and their cross-connectives. Immunoreactivity was evident also in the extensive, peripheral network of fine fibers of the subtegumental region and in the plexus of varicose fibers that innervate the muscle in each of the suckers. In dividing stages of the tetrathyridium, the immunoreactive lateral nerve cords of adjoining progeny were in continuity around the base of the division cleft.

Animals↗
PubMed1993

The primary structure of pancreatic polypeptide from a primitive insectivorous mammal, the European hedgehog (Erinaceous europaeus).

Pancreatic polypeptide (PP) has been isolated from extracts of the pancreas of the European hedgehog (Erinaceous europaeus) which is a representative of the order Insectivora, deemed to be the most primitive group of placental mammals. Pancreatic tissues were extracted in acidified ethanol and the peptide was purified chromatographically using a PP C-terminal hexapeptide amide specific radioimmunoassay to monitor purification. Two major PP-immunoreactive peptides were baseline-resolved following the final analytical reverse phase HPLC fractionation. Each was separately subjected to plasma desorption mass spectroscopy (PDMS) and gas-phase sequencing. The molecular masses of each peptide were similar: (I) 4237.6 +/- 4 Da and (II) 4238.2 +/- 4 Da. The full primary structures of each peptide were deduced and these were identical: VPLEPVYPGDNATPEQMAHYAAELRRYINMLTRPRY. The peptides were deemed to be amidated due to their full molar cross-reactivity with the amide-requiring PP antiserum employed in radioimmunoassay. The molecular mass (4233.8 Da) calculated from the sequence was in close agreement with PDMS estimates and the reason for the different retention times of each peptide is unknown at present. Hedgehog PP exhibits only 2 unique amino acid substitutions, at positions 1 (Val) and 19 (His), when compared with other mammalian analogues.

Amino Acid Sequence↗
PubMed1993

Isolation and primary structure of a novel avian pancreatic polypeptide from five species of Eurasian crow.

Chicken pancreatic polypeptide is the prototype of the neuropeptide Y (NPY)/PP superfamily of regulatory peptides. This polypeptide was appended the descriptive term avian, despite the presence of some 8600 extant species of bird. Additional primary structures from other avian species, including turkey, goose and ostrich, would suggest that the primary structure of this polypeptide has been highly-conserved during avian evolution. Avian pancreatic polypeptides structurally-characterised to date have distinctive primary structural features unique to this vertebrate group including an N-terminal glycyl residue and a histidyl residue at position 34. The crow family, Corvidae, is representative of the order Passeriformes, generally regarded as the most evolutionarily recent and diverse avian taxon. Pancreatic polypeptide has been isolated from pancreatic tissues from five representative Eurasian species (the magpie, Pica pica; the jay, Garrulus glandarius; the hooded crow, Corvus corone; the rook, Corvus frugilegus; the jackdaw, Corvus monedula) and subjected to structural analyses. Mass spectroscopy estimated the molecular mass of each peptide as 4166 +/- 2 Da. The entire primary structures of 36 amino acid residue peptides were established in single gas-phase sequencing runs. The primary structures of pancreatic polypeptides from all species investigated were identical: APAQPAYPGDDAPVEDLLRFYNDLQQYLNVVTRPRY. The peptides were deemed to be amidated due to their full molar cross-reactivity with the amide-requiring PP antiserum employed. The molecular mass (4165.6 Da), calculated from the sequences, was in close agreement with mass spectroscopy estimates. The presence of an N-terminal alanyl residue and a prolyl residue at position 34 differentiates crow PP from counterparts in other avian species.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

Discussion paper: towards a systematic classification for regulatory peptides.

Advances in microsequencing technology have led to the elucidation of primary structures of peptidic messengers from many eukaryotic and prokaryotic life forms. Existing peptide nomenclature is based upon such factors as bioactivity, source of isolation (tissue or species), chemical attributes, acronyms derived from a combination of these factors, or by other arbitrary means. In order to overcome many of the problems arising from current nomenclature, a standardised classification scheme for peptidic messengers is proposed which utilises an alphanumeric code string analogous to EC enzyme classification to convey information on origin, chemistry and relatedness to other similar molecules. It is anticipated that the scheme outlined will provide the basis for the rational classification of new peptides.

Animals↗
PubMed1993

An immunocytochemical study of putative neurotransmitters in the metacercariae of two strigeoid trematodes from rainbow trout (Oncorhynchus mykiss).

Whole mounts of the metacercariae of Diplostomum sp. and Cotylurus 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 FMRFamide) 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.

Animals↗
PubMed1993

Electron immunogold labeling of regulatory peptide immunoreactivity in the nervous system of Moniezia expansa (Cestoda: Cyclophyllidea).

An electron immunogold-labeling technique was used in conjunction with a post-embedding procedure to demonstrate for the first time the ultrastructural distribution of the parasitic platyhelminth neuropeptide, neuropeptide F (NPF), in the nervous system of the cestode Moniezia expansa. Two axon types, distinguished by their populations of different-sized electron-dense vesicles, were identified. Immunogold labeling demonstrated an apparent homogeneity of PP, FMRFamide and NPF (M. expansa) antigenic sites throughout the larger dense-cored vesicles within the central nervous system. Triple labeling clearly demonstrated the co-localisation of immunoreactivities (IR) for NPF, PP and FMRFamide within the same dense-cored vesicles. The presence of NPF-IR within the vesicles occupying the perikaryon of the neuronal cell body indicated that the peptides had undergone post-translational C-terminal amidation prior to entering the axon. Antigen pre-absorption experiments using NPF prevented labeling with either PP or FMRFamide antisera, and the failure of these antisera to block NPF-IR supports the view that some, if not all, of the PP/FMRFamide-IR is due to NPF-like peptides.

Animals↗
PubMed1993

Neuropeptide F-immunoreactivity in the tetrathyridium of Mesocestoides corti (Cestoda: Cyclophyllidea).

The distribution pattern and subcellular localisation of neuropeptide F (NPF) immunoreactivity (IR) in the tetrathyridium stage of Mesocestoides corti were investigated by whole-mount immunocytochemistry in conjunction with confocal scanning laser microscopy (CSLM) and by immunoelectron microscopy using immunogold labeling. Using an antiserum directed to the C-terminal decapeptide amide (residues 30-39) of synthetic NPF (Moniezia expansa), CSLM revealed NPF-IR throughout the central and peripheral nervous systems of parental and dividing tetrathyridia. Ultrastructurally, gold labeling of NPF-IR was confined to the contents of the smaller of the two sizes of electron-dense neuronal vesicle identified.

Animals↗
PubMed1993

A cytochemical study of the nervous system of the proteocephalidean cestode, Proteocephalus pollanicola.

The localization and distribution of cholinergic, serotoninergic and peptidergic nerve elements in the proteocephalidean tapeworm, Proteocephalus pollanicola, have been investigated by enzyme histochemistry, and by an indirect immunofluorescence technique interfaced with confocal scanning laser microscopy. Cholinesterase (ChE) activity was localized in the major components of the central nervous system (CNS) and the peripheral nervous system (PNS), including the innervation of the reproductive structures of the worm. Serotoninergic (5-HT) nerves were found in the paired cerebral ganglia, transverse commissure and in the 10 longitudinal nerve cords. Antisera to 17 mammalian regulatory peptides and the invertebrate peptide FMRFamide have been used to explore the peptidergic nervous system of the worm. The most extensive immunostaining occurred with antisera raised to members of the neuropeptide Y superfamily, namely neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP). In all cases, intense immunoreactivity was found in numerous cell bodies and fibres of both the CNS and PNS, including the innervation of the reproductive apparatus. FMRFamide antisera stained the same structures to a comparable degree as those raised to the NPY superfamily. Cholinergic and peptidergic elements were much more prevalent within the CNS, while the serotoninergic nerve fibres tended to dominate in the PNS. The overlap obtained in staining patterns for the peptidergic and cholinergic components suggests that there may be a certain amount of co-localization of peptides with small-molecule transmitter substances in the same neurone. Weak staining for the tachykinin, substance P and for calcitonin gene-related peptide (CGRP) was confined to the major longitudinal nerve cords.

Animals↗
PubMed1993

Rabbit pancreatic polypeptide.

1. In almost all studies involving localization or quantitation of regulatory peptides, an essential prerequisite is the generation of specific antisera in rabbits. Despite this almost universal practice, the primary structures of some established regulatory peptides, such as pancreatic polypeptide (PP), of the rabbit, remain unknown. 2. Here we report the full primary structure of PP isolated from extracts of rabbit pancreas. 3. PP immunoreactivity was purified using an antiserum (PP 221) generated to the highly-conserved C-terminal hexapeptide amide of mammalian PP. A single molecular form of rabbit PP was consistently resolved during sequential chromatographic fractionations. 4. Automated Edman degradation established the full primary structure as: APPEPVYPGDDATPEQMAEYVADLRRYINMLTRPRY. The molecular mass derived from this sequence (4196.7 Da), was in full agreement with that determined by mass spectroscopy (4196 Da). The peptide was deemed to be C-terminally amidated due to its full molar crossreactivity with the amide-requiring PP antiserum employed. 5. When compared with all other known mammalian PP sequences, rabbit PP displays three unique substituted sites, Pro at position 3, Glu at position 19 and Val at position 21.

Amino Acid Sequence↗
PubMed1993

Distribution and immunochemical characteristics of neuropeptide F (NPF) (Moniezia expansa) - immunoreactivity in Proteocephalus pollanicola (Cestoda: Proteocephalidea).

1. Using immunocytochemical techniques and confocal scanning laser microscopy, the proteocephalidean cestode, Proteocephalus pollanicola from Lough Neagh pollan (Coregomus autumnalis) was examined for the presence of the native platyhelminth neuropeptide, neuropeptide F (NPF). 2. An antiserum specific for whole-molecule NPF (1-39) (Moniezia expansa) did not immunostain nerve processes in P. pollanicola. A C-terminally-directed NPF (30-39) (M. expansa) antiserum immunostained nerve fibres and cell bodies of both the central and peripheral nervous systems, including innervation associated with the female reproductive system. 3. The pattern of immunoreactivity was identical to that obtained using antisera to the C-terminal region of mammalian NPY-superfamily peptides and the invertebrate neuropeptide, FMRFamide. 4. Under radioimmunoassay conditions, only the C-terminally-directed NPF antiserum cross-reacted with the P. pollanicola peptide and detected 58.74 ng/g wet weight of NPF-1 R in extracts of the worm. 5. Chromatographic characterisation of P. pollanicola NPF-immunoreactivity indicated an apparent molecular weight of 4400-4700 Da, similar to that of NPF (M. expansa). 6. Further analytical HPLC characterisation identified two molecular forms of P. pallanicoda NPF-immunoreactivity, both of which had different retention times from those of NPF (M. expansa) and NPF (Artioposthia triangulata). 7. These data suggest that P. pollanicola possesses a neuropeptide which is homologous in its C-terminal region to NPF (M. expansa) but differs in its mid- to N-terminal region.

Amino Acid Sequence↗
PubMed1993

Choline acetyltransferase (ChAT) immunoreactivity in a sub-population of mammalian intestinal endocrine cells.

1. The distribution of choline acetyltransferase (ChAT) in rat and guinea-pig intestine has been analysed using an indirect immunofluorescence technique. 2. ChAT immunoreactivity was apparent in nerve fibres and cell bodies of the myenteric and submucous plexus and in fibres throughout the muscle coats and the mucosa. 3. Staining was also evident in a sub-population of mucosal endocrine cells in the small intestine, implying the existence of this enzyme and its product (acetylcholine) in these cells. 4. These data are consistent with previous observations on the distribution of ChAT activity in mammalian intestine.

Animals↗
PubMed1993

Use of specific antisera for the localisation and quantitation of leucokinin immunoreactivity in the nematode, Ascaris suum.

1. The leucokinins (LKs) are a group of eight related peptides isolated from the cockroach, Leucophaea maderae. 2. Antisera raised against LK-V, which were specific for the conserved mid to C-terminal region of the LKs, were used to immunostain the parasitic nematode Ascaris suum. 3. LK-IR was observed in neurons in the anterior nerve ring, retrovesicular ganglion, and ventral and dorsal nerve cords of the parasite. Immunostaining was specific in that it was abolished by preabsorption of the antiserum with different leucokinins. Some of the LK-immunoreactive neurons were identified on the basis of their morphological similarity with identified neurons in the free-living nematode Caenorhabditis elegans. 4. Immunoreactivity towards a number of other peptides, notably neuropeptide F, FMRF-amide, KGQELE and KELTAE, has previously been demonstrated in all LK-immunoreactive neurons, thus confirming the multiple-peptidergic nature of certain nematode nerves. 5. LK-IR was demonstrated and quantified in a number of tissues using RIA. Highest amounts were found in extracts of gut; LK-IR was also demonstrated in extracts of body wall, heads and tails, testes, ovaries and pseudocoelomic fluid. 6. The distribution of tissue LK-IR did not correlate with the amount of neuronal tissue in the samples. 7. Dilution experiments suggested that the LK-IR in the parasite is heterogeneous and that the peptide(s) in some tissues may not be analogous to the insect LKs. 8. The LK-IR in the parasite remains to be characterized; however, the results suggest that peptides related to the leucokinins may have a far wider phylogenetic distribution than has hitherto been thought.

Amino Acid Sequence↗
PubMed1993

Cytochemical observations on the nervous system of adult Corrigia vitta.

Adult Corrigia vitta (Trematoda: Dicrocoelidea) inhabit the pancreatic duct of the fieldmouse, Apodemus sylvaticus, where, in numbers, they may occlude the duct lumen and prevent the flow of pancreatic secretions. Enzyme histochemical and immunocytochemical techniques, in conjunction with confocal scanning laser microscopy, have been used to examine the localization and distribution of cholinergic, serotoninergic (5-HT, serotonin) and peptidergic components of the nervous system of the adult worm. All three classes of neuronal mediator showed a common pattern of staining, occurring throughout the central and peripheral nervous systems. Of the four peptide immunoreactivities (IR) demonstrated (pancreatic polypeptide (PP), peptide YY (PYY), substance P (SP), FMRFamide), PP-IR was the most predominant, occurring not only within the central ganglia and longitudinal nerve cords, but also in subtegumental plexuses and in fibres associated with the egg-forming apparatus. PYY and FMRFamide IRs were evident throughout the central and peripheral nervous systems; FMRFamide immunostaining, in particular, highlighted innervation of the ootype and immunoreactive cell bodies around the Mehlis' gland. Both SP- and 5-HT-IRs were restricted to the cerebral ganglia, ventral nerve cords and associated cell bodies. The distribution patterns of these peptides and 5-HT within the nervous system of C. vitta suggest they are likely to function as neuronal mediators. PP, PYY and FMRFamide may also serve in regulating egg production.

Animals↗
PubMed1993

The cholinergic, serotoninergic and peptidergic components of the nervous system of Moniezia expansa (Cestoda, Cyclophyllidea).

The central (CNS) and peripheral (PNS) nervous systems of the cyclophyllidean tapeworm, Moniezia expansa, were examined for the presence of cholinergic, serotoninergic and peptidergic elements using enzyme cytochemical and immunocytochemical techniques in conjunction with light and confocal scanning laser microscopy. Cholinesterase activity and 5-hydroxytryptamine- and regulatory peptide-immunoreactivities (IRs) were localized to the nerve fibres and cell bodies of all of the major neuronal components in the CNS of the worm, including the cerebral ganglia and connecting commissure, the 10 longitudinal nerve cords and associated transverse ring commissures. Although each of the 3 systems appeared well developed and comprised a significant portion of the nervous system, the serotoninergic constituent was the most highly developed, consisting of a vast array of nerve fibres and cell bodies distributed throughout the strobila of the worm. A close association of cholinesterase reactivity and peptide-IRs was evident throughout the CNS, indicating the possible co-localization of acetylcholine and neuropeptides. Within the PNS, cholinergic activity and serotoninergic- and peptidergic-IRs occurred in the subtegumental network of nerve fibres and somatic musculature. Although all 3 neurochemical elements were present in the acetabula, they were found in different nerve fibres; only cholinergic and peptidergic cell bodies were found. The common genital opening, vagina and ootype regions of the reproductive system displayed a rich innervation of all 3 types of neuronal populations. Within the peptidergic system, immunostaining with antisera raised to the C-terminus of the neuropeptide Y superfamily of peptides and the invertebrate peptides, neuropeptide F (M. expansa) and FMRFamide was the most prevalent. Limited positive-IR for substance P and neurokinin A were also recorded in the CNS of the worm.

Animals↗
PubMed1993

Immunocytochemical demonstration of neuropeptides in the central nervous system of the roundworm, Ascaris suum (Nematoda: Ascaroidea).

The localization and distribution of neuropeptides in the central nervous system of the pig roundworm, Ascaris suum, have been determined by an indirect immunofluorescence technique in conjunction with confocal microscopy. Antisera to 25 vertebrate peptides and two invertebrate peptides were used to screen the worm for immunoreactivity (IR). Immunostaining was obtained with antisera to pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), gastrin, cholecystokinin (CCK), substance P (SP), atrial natriuretic peptide (ANP), salmon gonadotropin-releasing hormone (SGnRH), mammalian gonadotropin-releasing hormone (MGnRH), chromogranin A (CGA) and FMRFamide. The most extensive patterns of IR occurred with antisera to PYY, FMRFamide and gastrin. IR was evident in nerve cells and fibres in the ganglia associated with the anterior nerve ring and in the main nerve cords and their commissures; IR to FMRFamide also occurred in the posterior nerve ring. Immunostaining for the other peptides was confined to the nerve cords, with the number of immunoreactive nerve fibres varying from peptide to peptide.

Animals↗