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1.
Ideggyogy Sz ; 60(3-4): 89-93, 2007 Mar 30.
Article in English | MEDLINE | ID: mdl-17451046

ABSTRACT

Neurons expressing VIP/PHI precursor mRNA have been localized in the interstitial nucleus of Cajal. Unilateral surgical cut through the medial forebrain bundle failed to influence VIP/PHI mRNA expression in the Cajal nucleus while brainstem hemisection or unilateral transection of the medial longitudinal fascicle reduced it markedly, ipsilateral to the knife cuts. Thus, in contrast to forebrain projecting VIP neurons in the rostral periaqueductal gray, VIP/PHI neurons in the Cajal nucleus project downwards, to the lower brainstem.


Subject(s)
Coiled Bodies , Neurons , Peptide PHI/analysis , Vasoactive Intestinal Peptide/analysis , Animals , Coiled Bodies/chemistry , Female , Male , Neurons/chemistry , RNA, Messenger/analysis , Rats , Rats, Sprague-Dawley
2.
J Comp Neurol ; 453(3): 269-79, 2002 Nov 18.
Article in English | MEDLINE | ID: mdl-12378587

ABSTRACT

A monoclonal antibody directed at the multiphosphorylated epitope of axonal neurofilament-H (NF-H) was used to label axon-like fibers in the rabbit retina. NF-H-immunopositive fibers were found in the outer plexiform layer (OPL), inner plexiform layer (IPL), and optic fiber layer (OFL). The morphological characteristics of the labeled processes identified those in the OPL as horizontal cell axons and axon terminals and fibers in the OFL as axons of ganglion cells. The NF-H-positive profiles in the OPL formed a subset of horizontal cell processes labeled for calbindin. In the IPL, NF-H-immunoreactive profiles lay at all levels but were detected most often in the middle strata, 2-4. Occasionally, we observed NF-H-immuoreactive processes emerging from the IPL and entering either the GCL or the inner nuclear layer (INL). The labeled fibers in the IPL were typically very thin, less than 1 microm in diameter, and could often be followed for over 1 mm as they ran laterally across the retina. Cell bodies were never labeled by the immunoserum. To identify the NF-H-immunopositive fibers in the IPL, standard immunocytochemical double-labeling techniques were applied, using antibodies directed against several neurotransmitters or modulators thought to be expressed by axon-bearing amacrine cells. The NF-H-positive processes in the IPL were found to correspond to those labeled for tyrosine hydroxylase, somatostatin, substance P, and NADPH diaphorase activity. However, the NF-H labels did not colocalize with those against the vasoactive intestinal peptide-associated protein PHM27. Our results indicate that putative axons in the retina possess the multiphosphorylated NF-H protein found within classic axons in the central nervous system. These results thus support the idea that certain subtypes of amacrine and horizontal cells maintain true axons in the mammalian retina.


Subject(s)
Amacrine Cells/chemistry , Axons/chemistry , Neurofilament Proteins/analysis , Rabbits/metabolism , Amacrine Cells/enzymology , Animals , Antibodies, Monoclonal , Antibody Specificity , Axons/enzymology , Calbindins , Choline O-Acetyltransferase/analysis , Choline O-Acetyltransferase/immunology , NADPH Dehydrogenase/analysis , NADPH Dehydrogenase/immunology , Neurofilament Proteins/immunology , Peptide PHI/analysis , Peptide PHI/immunology , Retinal Ganglion Cells/chemistry , Retinal Ganglion Cells/enzymology , S100 Calcium Binding Protein G/analysis , S100 Calcium Binding Protein G/immunology , Somatostatin/analysis , Somatostatin/immunology , Substance P/analysis , Substance P/immunology , Tyrosine 3-Monooxygenase/analysis , Tyrosine 3-Monooxygenase/immunology
3.
Histochem J ; 32(5): 295-301, 2000 May.
Article in English | MEDLINE | ID: mdl-10939517

ABSTRACT

The colocalization of regulatory peptide immunoreactivities in endocrine cells of the chicken proventriculus at hatching has been investigated using the avidin-biotin technique in serial sections and double immunofluorescence in the same section for light microscopy, and double immunogold staining for electron microscopy. In addition to the eight immunoreactivities previously described in this organ, cells immunoreactive for peptide histidine isoleucine (PHI), peptide gene product 9.5 (PGP), and the amidating enzyme, peptidylglycine alpha-amidating monooxygenase (PAM) were observed. All the cells immunoreactive to glucagon were also immunostained by the PHI antiserum. In addition, all the glucagon-like peptide 1, avian pancreatic polypeptide, and some of the neurotensin-like cells costored also glucagon- and PHI-immunoreactive substances. PGP- and PAM-immunoreactivities were also found in the glucagon-positive cells. A small proportion of the somatostatin-containing cells were positive for PHI but not for other regulatory peptides. These results could suggest either the existence of a very complex regulatory system or that the endocrine system of the newborn chickens is not yet fully developed.


Subject(s)
Mixed Function Oxygenases/analysis , Multienzyme Complexes , Peptide PHI/analysis , Proventriculus/chemistry , Thiolester Hydrolases/analysis , Animals , Bombesin/immunology , Chickens , Glucagon/immunology , Glucagon-Like Peptide 1 , Mixed Function Oxygenases/immunology , Pancreatic Polypeptide/immunology , Peptide Fragments/immunology , Peptide PHI/immunology , Protein Precursors/immunology , Proventriculus/blood supply , Proventriculus/innervation , Proventriculus/ultrastructure , Serotonin/immunology , Thiolester Hydrolases/immunology , Ubiquitin Thiolesterase
4.
Microsc Res Tech ; 46(4-5): 257-64, 1999.
Article in English | MEDLINE | ID: mdl-10469462

ABSTRACT

The present article reviews the literature relative to VIP- and PHI-containing nerve fibers in the pineal gland of mammals. The article summarizes data on the presence and distribution of the two peptides in the brain of mammals, their role in neuronal metabolism, and the significance and origin of VIPergic and PHIergic cerebrovascular nerve fibers. Special emphasis is placed on VIP- and PHI-containing nerves in the pineal gland. The morphology of the fibers, the nature of the innervation, and the distribution of immunoreactive nerves within the pineal gland are examined. The review discusses the nature of the classical and "central" innervation of the pineal gland. The possible site of origin of pinealopetal VIPergic and PHIergic fibers is investigated, with special reference to ganglia of the head, and particularly to the pterygopalatine, otic, and trigeminal ganglia. The nature of VIP (and PHI) receptors is examined with reference to the most recent acquisitions in the field. Based on the data, a role for VIP (and PHI) in pineal metabolism is discussed.


Subject(s)
Peptide PHI/analysis , Pineal Gland/innervation , Vasoactive Intestinal Peptide/analysis , Amino Acid Sequence , Animals , Autonomic Nervous System/chemistry , Brain Chemistry , Humans , Molecular Sequence Data , Peptide PHI/physiology , Pineal Gland/physiology , Pituitary Gland/physiology , Receptors, Vasoactive Intestinal Peptide/analysis , Receptors, Vasoactive Intestinal Peptide/physiology , Swine , Vasoactive Intestinal Peptide/physiology
5.
Int J Parasitol ; 28(5): 825-9, 1998 May.
Article in English | MEDLINE | ID: mdl-9650063

ABSTRACT

Vasoactive intestinal peptide-like and peptide histidine isoleucine-like immunoreactivities were detected in the excretory duct of adult male and female Nippostrongylus brasiliensis, thus indicating the source of these two physiologically active peptides previously isolated from the excretory/secretory products of adult N. brasiliensis. In the nervous system immunoreactivity to both these peptides was confined to females and was found in the neurons of the ovijector associated ganglion. This is consistent with co-synthesis of vasoactive intestinal peptide-like and peptide histidine isoleucine-like peptides which has also been shown to occur in all mammalian vasoactive intestinal peptid-ergic neurons studied to date. However, in addition to this, and in common to some previous studies on helminth vasoactive intestinal peptide and peptide histidine isoleucine immunoreactivities, co-synthesis of the peptides was not indicated in a pair of branched neurons which projected posteriorly and peripherally from the ganglion associated with the ovijector of females and which terminated in two pairs of ganglia also exhibiting vasoactive intestinal peptide-like immunoreactivity only. The position of these ganglia indicated that they innervate muscles close to the body wall and may be responsible for the muscular contractions required for expulsion of eggs from female Nippostrongylus brasiliensis. This is also the first study to successfully detect these peptides in the excretory system of gastrointestinal nematodes.


Subject(s)
Neurons/chemistry , Neuropeptides/analysis , Nippostrongylus/chemistry , Peptide PHI/analysis , Vasoactive Intestinal Peptide/analysis , Animals , Female , Immunohistochemistry , Male , Microscopy, Confocal , Rats
6.
Cell Tissue Res ; 289(3): 427-37, 1997 Sep.
Article in English | MEDLINE | ID: mdl-9232821

ABSTRACT

The development of the retinohypothalamic tract (RHT) labeled with cholera toxin and of the vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine (PHI) systems was studied in the hypothalamus of neonatal mink by using immunohistochemistry. Retinal fibers were observed in the suprachiasmatic nucleus (SCN) from birth and were adult-like by day 14. VIP and PHI immunoreactivity was also present from birth. Melatonin binding was studied by autoradiography using [125I]melatonin as a ligand. A specific binding was detected in near-term fetal and neonate brains in the olfactory epithelium, various thalamic nuclei, the pineal gland, and the pars tuberalis of the pituitary gland, but never in the SCN. These results are discussed in the context of the potential role of daylight cycles and/or melatonin in entraining circadian rhythms in neonate mink.


Subject(s)
Melatonin/pharmacology , Peptide PHI/analysis , Retina/cytology , Suprachiasmatic Nucleus/cytology , Vasoactive Intestinal Peptide/analysis , Animals , Animals, Newborn , Autoradiography , Female , Immunohistochemistry , Iodine Radioisotopes , Melatonin/metabolism , Mink , Neurons/chemistry , Neurons/metabolism , Radioligand Assay , Retina/growth & development , Suprachiasmatic Nucleus/chemistry , Suprachiasmatic Nucleus/growth & development , Visual Pathways
7.
Arch Oral Biol ; 42(2): 137-47, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9134126

ABSTRACT

The occurrence and distribution of several neurochemical markers were investigated. Numerous nerve fibres were shown, using antibodies to protein gene product (PGP) 9.5, neurone-specific enolase, calcitonin gene-related peptide (CGRP), substance P. neurokinin A or protein S-100. The presence of vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine amide (PHI), neuropeptide tyrosine, dopamine-beta-hydroxylase (DBH), cholecystokinin/gastrin, glutamate and galanin was more scarce. Nerve fibres containing these above-mentioned markers were found at several locations, i.e. in the epithelium, connective tissue, and around blood vessels. In the taste buds, numerous PGP 9.5, neurone-specific enolase-, CGRP-, substance P-, neurokinin A- and protein S-100-containing structures were found, but few VIP and galanin ones. No immunoreactivity was found with antibodies against somatostatin, bombesin, enkephalin or dynorphin. These findings extend knowledge about the general as well as the neurochemical messenger-based innervation of rat fungiform papillae, forming a firm basis for future functional investigations of normal, experimental and also clinical materials.


Subject(s)
Nerve Tissue Proteins/analysis , Neuropeptides/analysis , Taste Buds/chemistry , Tongue/innervation , Animals , Calcitonin Gene-Related Peptide/analysis , Cholecystokinin/analysis , Dopamine beta-Hydroxylase/analysis , Fluorescent Antibody Technique, Indirect , Galanin/analysis , Gastrins/analysis , Glutamic Acid/analysis , Nerve Fibers/chemistry , Neurokinin A/analysis , Neuropeptide Y/analysis , Peptide PHI/analysis , Phosphopyruvate Hydratase/analysis , Rats , Rats, Sprague-Dawley , S100 Proteins/analysis , Substance P/analysis , Thiolester Hydrolases/analysis , Tongue/chemistry , Ubiquitin Thiolesterase , Vasoactive Intestinal Peptide/analysis
8.
Eur J Neurosci ; 9(12): 2613-23, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9517467

ABSTRACT

The rat suprachiasmatic nucleus (SCN) consists of several classes of neurons which can be identified by their transmitter content. Knowledge of putative interaction between these different cell types is essential in order to understand the possibilities of information processing within the SCN. The aim of the present study was therefore to obtain more information about the mutual innervation between the main cell classes in the rat SCN, viz. those containing the neuropeptides arginine vasopressin (AVP), vasoactive intestinal peptide (VIP), peptide histidine isoleucine (PHI), gastrin-releasing peptide (GRP) and somatostatin respectively. For this purpose, vibratome sections were double-immunolabelled for seven different peptide combinations and subsequently analysed by high-resolution confocal laser scanning fluorescence microscopy. Attention was focused on axosomatic appositions, the occurrence and frequency of which were quantitatively estimated. Our analysis of double-immunolabelled sections demonstrated that some of the VIP- and some of the GRP-immunoreactive nerve cells and endings showed colocalization. Assuming, on the basis of literature data, that VIP and PHI are always colocalized at the cellular level, the five main cell classes in the SCN appeared to be interconnected, at least axosomatically, in the following reciprocal way: AVP <--> VIP/PHI, AVP <--> GRP, AVP <--> somatostatin, somatostatin <--> VIP/PHI, somatostatin <--> GRP, VIP/PHI <--> GRP, VIP/PHI/GRP <--> GRP, VIP/PHI/GRP <--> VIP/ PHI. In addition to this heterologous axosomatic innervation, these cell groups also showed substantial homologous innervation. Supported by electron microscope data from the literature showing the existence of axodendritic synapses for some of these peptide combinations, our findings strongly suggest that the rat SCN comprises a complex synaptic network with strong interactive capabilities, which is probably a requisite for its biological clock function.


Subject(s)
Arginine Vasopressin/analysis , Neurons/chemistry , Neuropeptides/analysis , Suprachiasmatic Nucleus/chemistry , Suprachiasmatic Nucleus/cytology , Animals , Antibody Specificity , Arginine Vasopressin/immunology , Fluorescent Antibody Technique , Gastrin-Releasing Peptide/analysis , Gastrin-Releasing Peptide/immunology , Male , Microscopy, Confocal , Neurons/immunology , Neuropeptides/immunology , Peptide PHI/analysis , Peptide PHI/immunology , Rats , Rats, Wistar , Somatostatin/analysis , Somatostatin/immunology , Vasoactive Intestinal Peptide/analysis , Vasoactive Intestinal Peptide/immunology
9.
Histol Histopathol ; 11(4): 1089-100, 1996 Oct.
Article in English | MEDLINE | ID: mdl-8930650

ABSTRACT

The recent discovery of nitric oxide (NO) as a biological messenger molecule with unique characteristics has opened a new field in pineal research. This free radical gas is synthesized by the enzyme nitric oxide synthase (NOS) from L-arginine. The activation of adrenoreceptors in the membrane of the pinealocytes mediates the increase in NO through a mechanism that involves G proteins. In the pinealocyte, NO stimulates guanylyl cyclase resulting in an increased intracellular content of cGMP. The role of cGMP in pineal metabolism, however, is still enigmatic. Using enzyme histochemistry and immunohistochemistry, the presence of NOS has been confirmed in the pineal gland of some species. In the rat and especially in the sheep, NOS is located in nerve fibres innervating the gland. These nerve fibres also contain the neuropeptides vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI), and are probably of parasympathetic origin. In cell cultures and tissue sections NOS immunoreactivity has been shown to be present in pinealocytes of the rat and bovine but not in the sheep. Finally, NOS is also present in the endothelial cells of the blood vessels of the pineal gland. Accordingly, in the mammalian pineal gland, NO is synthesized in both presynaptic nerve fibers and pinealocytes, as well as in blood vessels. However, the anatomical location of NO synthesis varies considerably among species. NO released in the pineal gland, might influence both the pineal metabolism and the blood flow of the gland.


Subject(s)
Nitric Oxide Synthase/metabolism , Pineal Gland/enzymology , Animals , Cattle , Cyclic GMP/metabolism , Histocytochemistry , Homeostasis , Humans , Immunohistochemistry , Isoenzymes/analysis , Isoenzymes/metabolism , Mammals , Models, Neurological , Nerve Fibers/enzymology , Nerve Fibers/ultrastructure , Peptide PHI/analysis , Pineal Gland/cytology , Pineal Gland/innervation , Rats , Sheep , Vasoactive Intestinal Peptide/analysis
10.
J Comp Neurol ; 372(1): 1-8, 1996 Aug 12.
Article in English | MEDLINE | ID: mdl-8841917

ABSTRACT

Two groups of four rats each received a 15-minute light stimulus during the first part of the night (ZT14) and the second part (ZT19), respectively. After 45-60 minutes, the animals were killed by perfusion fixation. Adjacent Vibratome sections through the suprachiasmatic nucleus (SCN) were double-immunostained for the presence of peptide histidine isoleucine (PHI), gastrin releasing peptide (GRP) or vasoactive intestinal peptide (VIP) with Fos by using fluorophore-conjugated secondary antibodies. A few sections were triple-immunostained for PHI, GRP or VIP with vasopressin (VP) and Fos. Sections were analyzed with a confocal laser scanning microscope. It turned out that the ZT19 light stimulus induced 4.2 times more nuclear profiles in the SCN immunoreactive for Fos than the light stimulus given at ZT14. The SCN of control animals did not show any Fos immunoreactivity. After the ZT14 light stimulus, approximately 33% of the Fos profiles showed colocalization with a perikaryal profile immunoreactive for PHI, GRP or VIP, whereas at ZT19, this percentage had doubled to approximately 65%. After the light stimulus at ZT14, the relatively low Fos induction was numerically and proportionally most prominent in the PHI-immunoreactive perikarya. As compared with ZT14, the increase of Fos after the ZT19 light stimulus was most pronounced in the GRP-immunoreactive perikarya (21x) followed by VIP (15x) and PHI (5x). This outcome suggests that at least three different cell groups characterized by, respectively, PHI alone, GRP, and VIP fully or partly colocalized with PHI, play a prominent role during light-induced phase shifts: the PHI neurons during light-induced phase delays, the GRP and VIP/(PHI) neurons during light-induced phase advances.


Subject(s)
Circadian Rhythm/physiology , Nerve Tissue Proteins/analysis , Neurons/chemistry , Neuropeptides/analysis , Proto-Oncogene Proteins c-fos/analysis , Suprachiasmatic Nucleus/chemistry , Animals , Gastrin-Releasing Peptide , Immunohistochemistry , Male , Peptide PHI/analysis , Peptides/analysis , Photic Stimulation , Rats , Rats, Wistar , Suprachiasmatic Nucleus/cytology , Suprachiasmatic Nucleus/radiation effects , Vasoactive Intestinal Peptide/analysis , Vasopressins/analysis
11.
Peptides ; 16(2): 275-87, 1995.
Article in English | MEDLINE | ID: mdl-7540293

ABSTRACT

The peptidergic innervation of the human superficial temporal artery was investigated by means of immunohistochemical, ultrastructural, and in vitro pharmacological techniques. A dense network of nerve fibers was found in the adventitia. The majority of the nerve fibers displayed immunoreactivity for tyrosine hydroxylase and neuropeptide Y (NPY). A moderate supply of perivascular nerve fibers displayed either acetylcholinesterase activity or immunoreactivity for vasoactive intestinal peptide (VIP), peptide histidine methionine-27 (PHM), and calcitonin gene-related peptide (CGRP). Only a few nerve fibers displayed substance P (SP), neurokinin A (NKA), and neuropeptide K (NPK) immunoreactivity. In double immunostained preparations, SP immunoreactivity was co-localized with NPK and CGRP in the same nerve fibers. Ultrastructural studies revealed the presence of numerous axon variocosities at the adventitial--medial border. NPY, VIP, and CGRP immunoreactivities occurred in the same type of large granular vesicles, but in morphological distinct nerve profiles. NPY had, in general, no direct vasoconstrictor effect. However, at a low concentration of NPY contractile response induced by NA (10(-7)-10(-6)M) was 9-15 times enhanced. The NPY-induced potentiation of the NA-induced contraction was not dependent on the presence of an intact endothelium. No significant difference was found between acetylcholine, VIP, and PHM in either potency or degree of relaxation. SP, NKA, and CGRP also acted as vasodilatory agents, with CGRP being more potent than the tachykinins. The response to SP, but not CGRP, was dependent on an intact endothelium. Pretreatment of the vessels with a low concentration of NPY did not change the responses to ACh, VIP, SP, or CGRP.


Subject(s)
Nerve Fibers/ultrastructure , Tachykinins , Temporal Arteries/innervation , Acetylcholinesterase/analysis , Animals , Antibodies , Calcitonin Gene-Related Peptide/analysis , Fluorescent Antibody Technique , Humans , Microscopy, Electron , Microscopy, Immunoelectron , Muscle Contraction , Muscle, Smooth, Vascular/innervation , Muscle, Smooth, Vascular/physiology , Muscle, Smooth, Vascular/ultrastructure , Neurokinin A/analysis , Neuropeptide Y/analysis , Neuropeptides/analysis , Peptide PHI/analysis , Rabbits/immunology , Rats/immunology , Substance P/analysis , Temporal Arteries/physiology , Temporal Arteries/ultrastructure , Tyrosine 3-Monooxygenase/analysis , Vasoactive Intestinal Peptide/analysis
12.
Clin Exp Dermatol ; 19(6): 463-72, 1994 Nov.
Article in English | MEDLINE | ID: mdl-7534221

ABSTRACT

In this study radioimmunoassay was used to determine neuropeptide levels in extracts from 17 differing anatomical regions of human skin. Marked regional variations of neuropeptide content for human skin were found and these variations are likely to reflect true physiological functions for the neuropeptides studied. In general the tachykinins, substance P (SP), neurokinin A (NKA) and calcitonin gene-related peptide (CGRP) were found in highest concentrations in regions of skin with the greatest tactile sensation. By contrast, highest concentrations of vasoactive intestinal peptide (VIP) and peptide histidine methionine (PHM) were found in axillary skin, where they probably play a part in axillary eccrine sweat production. Neurotensin was not found in any of the skin areas sampled, suggesting that it is relatively unimportant in human physiological skin control. Reverse-phase high-performance liquid chromatography (rpHPLC) was used to verify the results of radioimmunoassay. Both SP and NKA occurred in several regions in both their reduced and oxidized forms, as well as displaying molecular heterogeneity. CGRP occurred as one molecular species, this being alpha-CGRP, suggesting that this is the predominant molecular form in human skin. Likewise, both VIP and PHM displayed molecular homogeneity in the regions investigated by rpHPLC.


Subject(s)
Neuropeptides/analysis , Skin/chemistry , Adult , Aged , Calcitonin Gene-Related Peptide/analysis , Chromatography, High Pressure Liquid , Female , Humans , Male , Middle Aged , Neurokinin A/analysis , Peptide PHI/analysis , Radioimmunoassay , Substance P/analysis , Vasoactive Intestinal Peptide/analysis
13.
J Chem Neuroanat ; 7(3): 171-84, 1994 Aug.
Article in English | MEDLINE | ID: mdl-7531454

ABSTRACT

By means of indirect immunofluorescence the neuropeptides somatostatin, galanin and peptide histidine isoleucine were localized in cell bodies, nerve fibres and terminal-like elements in the ganglion and spinal nucleus of the human trigeminal nerve in perinatal and adult ages. No immunoreactivity to vasoactive intestinal polypeptide was observed. In the gasserian ganglion somatostatin-, galanin- and peptide histidine isoleucine-containing neurons and nerve fibres occurred frequently in pre- and full-term newborns, but were scarce to absent in adults. Somatostatin- and galanin-positive pericellular basket-like structures around non-immunoreactive perikarya were observed in newborn specimens. Immunoreactivity to somatostatin, galanin and peptide histidine isoleucine labelled nerve fibers and punctate and felt-like nerve terminals in the pars interpolaris and subnucleus caudalis of the spinal trigeminal nucleus, with immunostaining and distribution patterns characteristic for each peptide. In addition, somatostatin-containing neuronal cell bodies frequently were detected. At variance with those containing somatostatin, the number of galanin- and peptide histidine isoleucine-like immunoreactive elements were dramatically reduced in the adult tissue compared to the newborn one. Double immunostaining revealed that each of the three peptides partially colocalizes with substance P, the degree of coexistence being very low for somatostatin/substance P and high for galanin/substance P and peptide histidine isoleucine/substance P both in the gasserian ganglion and in the spinal nucleus. The results obtained suggest that somatostatin, galanin and peptide histidine isoleucine may play functional roles in primary sensory neurons and at the first synaptic level of the human trigeminal sensory system.


Subject(s)
Infant, Newborn/metabolism , Neurons/chemistry , Neuropeptides/analysis , Substance P/analysis , Trigeminal Ganglion/chemistry , Trigeminal Nucleus, Spinal/chemistry , Aged , Female , Galanin , Humans , Immunohistochemistry , Male , Middle Aged , Peptide PHI/analysis , Peptides/analysis , Somatostatin/analysis
14.
Hum Reprod ; 9(6): 1053-7, 1994 Jun.
Article in English | MEDLINE | ID: mdl-7962375

ABSTRACT

Vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM) originate from the same precursor molecule, prepro VIP. In the present study we examined the concentrations of VIP and PHM in human follicular fluid and their effects on cultured human granulosa/lutein cells. Follicular fluid and cells were obtained from patients undergoing in-vitro fertilization for tubal infertility. The concentrations of VIP and PHM in pre-ovulatory human follicular fluid were measured radioimmunochemically. Granulosa/lutein cells isolated from follicular fluid were cultured under serum-free conditions with VIP and PHM in varying concentrations (0.1, 10, 1000 nmol/l). [3H]Thymidine incorporation in the cells and oestradiol as well as progesterone concentrations in the culture medium were measured. The mean (+/- SEM) concentrations of VIP and PHM were 6.8 +/- 0.1 and 7.7 +/- 0.8 pmol/l, respectively. VIP at a concentration of 10 nmol/l caused a significant increase in [3H]thymidine incorporation, and at 1000 nmol/l a significant increase in oestradiol secretion was observed. VIP had no effect on progesterone secretion. PHM at the concentrations tested did not influence any of the activities. We conclude that VIP and PHM are present in human preovulatory follicular fluid and that VIP stimulates DNA synthesis and oestradiol secretion in cultured human granulosa/lutein cells. This indicates that VIP and perhaps PHM participate in the local nervous regulation of human ovarian function.


Subject(s)
DNA/biosynthesis , Follicular Fluid/chemistry , Peptide PHI/analysis , Steroids/physiology , Vasoactive Intestinal Peptide/analysis , Cell Division/drug effects , Cells, Cultured , Female , Granulosa Cells/drug effects , Granulosa Cells/metabolism , Humans , Luteal Cells/drug effects , Luteal Cells/metabolism , Peptide PHI/pharmacology , Vasoactive Intestinal Peptide/pharmacology
15.
Gen Comp Endocrinol ; 93(3): 411-23, 1994 Mar.
Article in English | MEDLINE | ID: mdl-7910805

ABSTRACT

The distributions of tyrosine hydroxylase (TH), protein gene product (PGP) 9.5, calcitonin gene-related peptide (CGRP), and peptide histidine isoleucine (PHI) have been examined immunohistochemically in the adrenal gland of the snake Waglerophis merremii. The morphology of chromaffin cells and the presence of ganglionic neurons in the gland revealed by means of the glutaraldehyde-silver technique and electron microscopy are also described. Two distinct types of TH-immunoreactive (-IR) cells are present in the dorsal noradrenergic ribbon: small chromaffin cells and a larger type identified as ganglionic neurons. Small, mostly round or fusiform cells often displayed long processes. Ganglionic cells, arranged in patches, had long processes entering the cortex of the gland. Chromaffin adrenergic cells, forming small groups of 4-7 cells, were scattered within the interrenal tissue and had a wide variety of shapes with processes that appeared to contact other chromaffin cells. Bundles of PGP 9.5-IR fibers occurred in the subcapsular zone of the adrenal gland with fibers entering the cortex and dorsal noradrenergic ribbon of the gland. Thick and thin TH-IR fibers were seen. Thick TH-IR fibers were nonvaricose and appeared to originate mainly in ganglionic neurons. Thin TH-IR fibers with small varicosities were numerous in the interrenal tissue and were frequently seen between clusters of adrenergic cells in close apposition to cortical cells and vessels. CGRP-IR fibers were present throughout the entire adrenal gland, whereas PHI-IR fibers had a preferential distribution in the interrenal tissue. Both CGRP- and PHI-IR fibers were closely associated with vessels and cortical cells.


Subject(s)
Adrenal Glands/chemistry , Chromaffin System/cytology , Nerve Fibers/ultrastructure , Snakes/anatomy & histology , Adrenal Glands/ultrastructure , Animals , Calcitonin Gene-Related Peptide/analysis , Chromaffin System/chemistry , Female , Fluorescent Antibody Technique , Glutaral , Immunohistochemistry , Male , Microscopy, Electron , Nerve Fibers/chemistry , Peptide PHI/analysis , Silver , Snakes/metabolism , Thiolester Hydrolases/analysis , Tyrosine 3-Monooxygenase/analysis , Ubiquitin Thiolesterase
16.
Exp Eye Res ; 58(1): 117-26, 1994 Jan.
Article in English | MEDLINE | ID: mdl-8157097

ABSTRACT

The origin and neuropeptide content of nerve fibres in the rat conjunctiva were studied by retrograde tracing and denervations in combination with immunocytochemistry. Immunocytochemistry revealed nerve fibres containing neuropeptide Y (NPY), vasoactive intestinal peptide (VIP), peptide histidine isoleucine (PHI), helospectin, substance P (SP), and calcitonin gene-related peptide (CGRP) around blood vessels, smooth muscle bundles and glands. Removal of the sphenopalatine ganglion eliminated the majority of the VIP-, PHI- and helospectin-containing fibres. Sympathectomy eliminated the majority of the NPY-containing fibres in the conjunctiva. Removal of both the sphenopalatine ganglion and the superior cervical ganglion led to further reduction of the NPY fibres. Injection of the retrograde tracer True Blue into the conjunctiva resulted in the appearance of labelled cell bodies in the trigeminal ganglion, the sphenopalatine ganglion, the superior cervical ganglion, and the ciliary ganglion. Judging from the number of labelled nerve cell bodies, the superior cervical ganglion contributes most to the innervation. These results indicate that the majority of NPY-containing nerve fibres in the conjunctiva derives from the superior cervical ganglion (sympathetic nerve supply). Most of the VIP- and a minor population of the NPY-containing fibres in the conjunctiva originate in the sphenopalatine ganglion (parasympathetic nerve supply). A sparse amount of VIP- and NPY-containing fibres derives from the ciliary ganglion. Nerve fibres containing SP and/or CGRP emanate from the trigeminal ganglia (sensory nerve supply). Together the findings indicate that several ganglia project to the conjunctiva and that many neuropeptides may be involved in the control of conjunctival activity.


Subject(s)
Conjunctiva/innervation , Neuropeptides/analysis , Animals , Calcitonin Gene-Related Peptide/analysis , Denervation , Ganglia, Parasympathetic/chemistry , Immunohistochemistry , Inclusion Bodies/chemistry , Male , Neural Pathways , Neurons/chemistry , Neuropeptide Y/analysis , Peptide PHI/analysis , Rats , Rats, Wistar , Superior Cervical Ganglion/chemistry , Trigeminal Ganglion/chemistry , Vasoactive Intestinal Peptide/analysis
17.
Eur J Neurosci ; 5(11): 1510-9, 1993 Nov 01.
Article in English | MEDLINE | ID: mdl-7506974

ABSTRACT

Using immunohistochemistry and in situ hybridization, we studied changes in expression of some neuropeptides in large and medium-sized neurons in lumbar 4 and 5 rat dorsal root ganglia projecting to the gracile nucleus, in response to peripheral axotomy. Fourteen days after unilateral sciatic nerve transection, many large neurons and some medium-sized neurons in ipsilateral dorsal root ganglia were strongly neuropeptide Y-positive. Galanin-, vasoactive intestinal polypeptide (VIP)- and peptide histidine-isoleucine (PHI)-like immunoreactivities coexisted with neuropeptide Y-like immunoreactivity in some of these neurons. After axotomy numerous large and medium-sized cells contained neuropeptide Y mRNA in the ipsilateral ganglia, whereas no hybridization was seen in the contralateral or control ganglia. Cross-sectioned, large neuropeptide Y-positive fibres were observed in a somatotopically appropriate zone within the ipsilateral gracile fasciculus. A dense network of neuropeptide Y-immunoreactive, large nerve fibres and terminals was seen in the ipsilateral gracile nucleus. A small number of galanin- and VIP/PHI-like immunoreactive nerve fibres and terminals were also observed in adjacent sections. Neuropeptide Y-like immunoreactivity colocalized with galanin- or VIP/PHI-like immunoreactivity in some nerve fibres. None of these neuropeptide immunoreactivities could be detected in nerve fibres and terminals in the control or contralateral gracile nucleus. These findings suggest that neuropeptides, in addition to their role in small dorsal root ganglion neurons, may have a function in large and medium-sized dorsal root ganglion neurons projecting to laminae III and IV in the dorsal horn as well as to the gracile nuclei, as a part of their response to peripheral axotomy.


Subject(s)
Afferent Pathways/metabolism , Ganglia, Spinal/metabolism , Neurons/metabolism , Neuropeptide Y/biosynthesis , Sciatic Nerve/physiology , Animals , Fluorescent Antibody Technique , Galanin , Gene Expression , Immunohistochemistry , In Situ Hybridization , Male , Neuropeptide Y/analysis , Neuropeptide Y/metabolism , Neuropeptides/analysis , Neuropeptides/metabolism , Oligonucleotide Probes , Peptide PHI/analysis , Peptide PHI/metabolism , Peptides/analysis , Peptides/metabolism , Rats , Rats, Sprague-Dawley , Vasoactive Intestinal Peptide/analysis , Vasoactive Intestinal Peptide/metabolism
18.
Acta Histochem ; 94(2): 197-201, 1993 May.
Article in English | MEDLINE | ID: mdl-7688926

ABSTRACT

The innervation of the human male distal urethra and intrinsic external urethral sphincter was investigated by immunohistochemical techniques. The innervation density was low. In the urethra neuropeptide, and vasoactive intestinal polypeptide positive nerves were found in the muscular layer, calcitonin gene-related peptide, galanin and PHI nerves were localized in the epithelium. In the intrinsic external urethral sphincter NPY and galanin nerves were quite numerous between the striated muscle bundles, calcitonin gene-related peptide, vasoactive intestinal polypeptide and PHI were only seen occasionally as thin fibres.


Subject(s)
Urethra/innervation , Calcitonin Gene-Related Peptide/analysis , Galanin , Humans , Immunohistochemistry , Male , Neuropeptide Y/analysis , Neuropeptides/analysis , Peptide PHI/analysis , Peptides/analysis , Urethra/chemistry , Vasoactive Intestinal Peptide/analysis
19.
Cancer ; 71(9): 2841-6, 1993 May 01.
Article in English | MEDLINE | ID: mdl-7682155

ABSTRACT

BACKGROUND: It has been reported that vasoactive intestinal peptide (VIP)-producing tumors accompanied by watery diarrhea, hypokalemia, and achlorhydria (WDHA) syndrome often produce multiple hormones biochemically and immunohistochemically. METHODS: The authors examined the distribution of several peptides--VIP, peptide histidine methionine (PHM), neuropeptide Y (NPY), methionine-enkephalin (M-EK), somatostatin (SS), substance-P (Sub-P), corticotropin-releasing hormone, and tyrosine hydroxylase--with immunohistochemical studies and an in situ hybridization method in three ganglioneuroblastomas with WDHA syndrome. All patients had an extremely elevated plasma level of VIP. RESULTS: Peptides examined immunohistochemically in the tumor were all detectable in ganglionic cells and some neurites. The coexistence of those peptides was observed with serial section staining. The presence of messenger RNA of VIP/PHM-27, NPY, and SS was detectable in the cytoplasm of the tumor cells with the in situ hybridization. CONCLUSIONS: It was shown that multiple genes of peptides are expressed simultaneously and translated to proteins in those tumors.


Subject(s)
Ganglioneuroma/genetics , Gene Expression Regulation, Neoplastic , Retroperitoneal Neoplasms/genetics , Vipoma/genetics , Child, Preschool , Corticotropin-Releasing Hormone/analysis , Enkephalin, Methionine/analysis , Female , Gene Expression Regulation, Neoplastic/genetics , Humans , In Situ Hybridization , Infant , Male , Neuropeptide Y/analysis , Peptide PHI/analysis , Somatostatin/analysis , Substance P/analysis , Tyrosine 3-Monooxygenase/analysis , Vasoactive Intestinal Peptide/analysis
20.
Res Vet Sci ; 54(3): 335-9, 1993 May.
Article in English | MEDLINE | ID: mdl-7687789

ABSTRACT

The tone of respiratory smooth muscle is largely determined by the input from autonomic nerves. The distribution of adrenergic and selected nonadrenergic, non-cholinergic (NANC) nerves in the normal equine trachealis muscle was investigated using immunohistochemistry. The smooth muscle of the trachealis was found to contain numerous nerves immunoreactive for an enzymatic marker of adrenergic nerves, as well as many nerves immunoreactive for a putative NANC neurotransmitter, peptide histidine isoleucine, a potent bronchodilator. The tissue surrounding the respiratory smooth muscle contained numerous nerves immunoreactive for the neuropeptides substance P and calcitonin gene-related peptide, which can cause marked vasodilation and bronchoconstriction. The complex innervation of the equine trachea should be kept in mind when interpreting the results of physiological experiments.


Subject(s)
Autonomic Nervous System/anatomy & histology , Horses/anatomy & histology , Muscle, Smooth/innervation , Nerve Fibers/ultrastructure , Neuropeptides/analysis , Peptide PHI/analysis , Trachea/innervation , Animals , Autonomic Nervous System/cytology , Biomarkers/analysis , Calcitonin Gene-Related Peptide/analysis , Dopamine beta-Hydroxylase/analysis , Immunohistochemistry , Substance P/analysis
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