Galanin, substance P and calcitonin gene-related peptide are colocalized in some afferent neurons innervating the porcine ovary.
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Animals↗
Biomedical subjects
M Lakomy
Publications and source records attributed to M Lakomy.
Peptidergic innervation of the porcine oviduct studied by double-labelling immunohistochemistry.
Neurochemical coding of nerve fibres supplying the porcine oviduct was studied by means of double-labelling immunofluorescence. Immunoreactivities to rate-limiting enzymes of catecholamine synthesis, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DbetaH), were found in numerous oviductal nerve fibres. All TH-immunoreactive (TH-IR) nerve processes were simultaneously DbetaH-IR. This subset of nerves was considered to be sympathetic noradrenergic. In addition to noradrenaline, many axons additionally exhibited immunoreactivity to neuropeptide Y (NPY), or leu5-enkephalin (Leu-ENK). Small numbers of somatostatin- (SOM-) or vasoactive intestinal polypeptide-immunoreactive (VIP-IR) fibres, sometimes coexpressing TH/DbetaH-immunoreactivity, supplied the porcine oviduct. Substance P- (SP- ) and/or calcitonin gene-related peptide-immunoreactive (CGRP-IR) nerve fibres were only sporadically found. Although these nerves did not contain TH/DbetaH-immunoreactivity, they often ran in close vicinity to TH/DbetaH-IR axons, forming together thin nerve bundles. All the above mentioned subpopulations of nerve fibres were found throughout the entire length of the oviduct being mainly related to the vascular and non-vascular smooth myocytes. However, some of the putative afferent (i.e. SP- or CGRP-IR) or parasympathetic efferent (i.e. VIP- or NPY-IR but TH/DbetaH-immunonegative) axons were located beneath the epithelium. Such distribution implies these nerve fibres to be involved in the regulation of the oviductal blood flow, non-vascular smooth myocyte tonus, transmission of sensory information and control of the epithelial secretion.
Animals↗
5-Hydroxytryptamine-immunoreactive nerve fibers in the rat and porcine prevertebral sympathetic ganglia: effect of precursor loading and relation to catecholaminergic neurons.
Localization of 5-hydroxytryptamine immunoreactivity was studied in the rat coeliac-superior mesenteric ganglion complex and in the porcine superior and inferior mesenteric ganglia by the indirect immunofluorescence technique. In normal rats, only 5-hydroxytryptamine immunoreactive SIF cells were seen in the coeliac-superior mesenteric ganglion complex. In the rats, pretreated with a 5-hydroxytryptamine precursor, L-tryptophan, and with a monoamine oxidase inhibitor, nialamide, a large number of 5-hydroxytryptamine-immunoreactive nerve fiber terminals were detected. In normal porcine superior and inferior mesenteric ganglia, intense 5-hydroxytryptamine immunoreactivity was found in numerous nerve fibers which were located around tyrosine hydroxylase-immunoreactive principal neurons. The origin and function of these fibers are discussed.
Animals↗
Distribution and immunohistochemical characteristics of neurons in the porcine caudal mesenteric ganglion projecting to the vas deferens and seminal vesicle.
Combined retrograde tracing (using fluorescent tracer Fast Blue) and double-labelling immunofluorescence were used to study the distribution and immunohistochemical characteristics of neurons in the porcine caudal mesenteric ganglion projecting to the vas deferens and seminal vesicle. The distribution and immunohistochemical properties of neurons projecting to both organs were similar. As revealed by retrograde tracing, Fast Blue-positive neurons were located within the left and right ganglia, with a distinct predominance in the ipsilateral one. In the ipsilateral ganglion, the majority of the neurons were located caudally, along the dorso-lateral ganglionic border, suggesting a somatotopic organization of the ganglion. Immunohistochemistry revealed four populations of retrogradely labelled neurons (from the largest to the smaller one): tyrosine hydroxylase-positive/neuropeptide Y-negative (TH+/NPY-), TH+/NPY+, TH-/NPY-, TH-/NPY+. With respect to their surrounding nerve fibres, two subpopulations of the dye-labelled neurons could be distinguished. The small one consisted of solitary neurons receiving a strong calcitonin gene-related peptide- and Leu5-enkephalin-, and a less intense vasoactive intestinal peptide-immunoreactive innervation. The remaining neurons were poorly supplied by singular nerve fibres containing some of the investigated peptides. We conclude that the caudal mesenteric ganglion should be considered as a prominent source of adrenergic and/or NPY-positive innervation for the porcine male reproductive tract.
Amidines↗
The distribution and co-localization of immunoreactivity to nitric oxide synthase, vasoactive intestinal polypeptide and substance P within nerve fibres supplying bovine and porcine female genital organs.
The distribution of nitric oxide synthase-immunoreactive (NOS-IR) axons and their relationship to structures immunoreactive to vasoactive intestinal polypeptide (VIP), substance P (SP) and tyrosine hydroxylase (TH) were studied by means of the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) technique or double-labelling immunofluorescence in the genital organs of cow and pig. Relevant neurons were also investigated in the pig. NOS-containing neural structures were TH-immunonegative in bovine or porcine genital organs, or in the studied ganglia. In the bovine ovary, NOS-IR nerves were neither VIP-IR nor SP-IR, whereas in the pig, most NOS-containing axons were also VIP-IR. The oviduct was supplied by single NOS/VIP- or NOS/SP-containing nerves, whereas in the uterus, NOS-IR axons were moderate in number, often being immunoreactive for VIP or SP. Numerous NOS/VIP-IR and NOS/SP-IR nerves were found in the vagina of both species. In all tissues studied, NOS-IR axons were mainly related to vascular smooth muscle. Most of the neurons of the paracervical ganglia and some neurons in dorsal root ganglia exhibited strong NOS activity. Only single neurons in sympathetic ganglia were NADPH-d-positive. Most nitrergic neurons in the autonomic ganglia were VIP-IR but SP-immunonegative. The sensory neurons were mostly NOS/SP-IR, whereas only single neurons co-expressed NOS and VIP immunoreactivity.
Animals↗
Existence and co-existence of vasoactive substances in nerve fibres supplying the abdomino-pelvic arterial tree of the female pig and cow.
The occurrence and co-localization of several presumed vasoactive neuropeptides, serotonin, and catecholamine-synthesising enzymes--tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (D beta H) and phenylethanolamine-N-methyltransferase (PNMT)--were investigated in perivascular nerves supplying the systemic and distributing arteries of the abdomino-pelvic arterial tree of the female pig and certain arteries supplying female reproductive organs in the cow. As revealed by single immunofluorescence, perivascular axons immunoreactive for TH, D beta H, neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP) and Leu-enkephalin (LENK) occurred in both species examined, whereas galanin-immunoreactive (GAL-IR) nerve fibres were found exclusively in the pig. PNMT-, serotonin-, dynorphin A-, alpha-neoendorphin-, bombesin- or cholecystokinin-IR nerve terminals were not observed. The following classes of perivascular nerve fibres might be distinguished in the present study: 1) noradrenergic (i.e. TH/D beta H-IR), 2) NPY-, 3) GAL- (pig only), 4) LENK-, 5) VIP-, 6) SP-, 7) VIP/NPY-, 8) SP/CGRP-, 9) SP/GAL- (pig only), 10) SP/VIP- (cow only), 11) TH/D beta H/NPY- and 12) TH/D beta H/NPY/LENK-IR. Distinct differences in the distribution of LENK- and SP-IR axons around particular parts of the studied arterial tree in individual species were also observed. The present data indicate that the abdomino-pelvic arterial tree of the pig and cow receive perivascular nerve fibres that exhibit diverse chemical codes, and that different chemical coding of perivascular nerve fibres in individual species may depend on the target organ of the particular artery.
Abdomen↗
Peptidergic innervation of the bovine vagina and uterus.
The distribution of neuropeptide Y, substance P, vasoactive intestinal polypeptide, Leu5-enkephalin, bombesin/gastrin-releasing peptide, calcitonin gene-related peptide, somatostatin, cholecystokinin and catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine-beta-hydroxylase was studied immunohistochemically in nerve fibres supplying the bovine vagina and uterus. The nerves containing tyrosine hydroxylase or dopamine-beta-hydroxylase and neuropeptide Y-immunoreactivity were particularly numerous in both organs. Substance P, vasoactive intestinal polypeptide and Leu5-enkephalin-containing nerves were less numerous whereas somatostatin and calcitonin gene-related peptide-immunoreactive nerves occurred occasionally. Bombesin/gastrin-releasing peptide and cholecystokinin immunoreactivities were not present in nervous fibers of the bovine uterus and vagina. Generally, the immunoreactive nerve terminals, fibers, networks or nerve bundles were present below the serous membrane, between smooth muscle cells of muscular layers, around blood vessels, in the submucosal layer and below the luminal epithelium of the uterus and cervix.
Animals↗
Immunohistochemical localization of galanin in bovine reproductive organs.
The vagina, uterus and oviduct were shown to receive galanin immunoreactive (GAL-IR) nerve fibres, the number of which varied between particular organs. In the ovary, GAL-IR nerves were absent. A small number of these nerves were located in the layers of the oviduct. A moderate number of GAL-IR nerves were situated in the body and uterine horns, whereas the uterine cervix and vagina wall contained a large number of GAL-IR nerve fibres, evenly distributed throughout particular membranes of the organs. GAL-IR nerves were found to contain, simultaneously, either vasoactive intestinal polypeptide (VIP), substance P (SP) or Leu5-enkephalin (ENK). Many of the GAL-IR nerves contained tyrosine hydroxylase (TH). A group of GAL-IR nerves that did not possess immunoreactivity to VIP, SP, ENK or TH was also observed.
Animals↗
NADPH-diaphorase-containing cerebrovascular nerve fibres and their possible origin in the pig.
NADPH-diaphorase histochemical technique was applied to demonstrate the catalytic activity of nitric oxide synthase (NOS) in the nerve fibers supplying some porcine cerebral blood vessels, as well as in ganglia thought to be their possible sites of origin. Five sexually mature Large With Polish race sows were used. The following blood vessels and their branches were studied: the basal cerebral artery, medial cerebral artery, arteries of the circle of Wills as well as arteries located in the arachnoidea. The activity of NADPH-d was visualised in whole-mount preparations from the above listed blood vessels. The presence of NADPH-d was additionally studied in the ganglia of trigeminal nerves, the sensory ganglia of vagus nerves, the pterygopalatine and cranial cervical ganglia. NADPH-d activity was found in nerve fibres supplying all the studied arteries. Larger blood vessels, the basal cerebral artery, medial cerebral artery and arteries of the circle of Wills possessed very dense NADPH-d-positive nerve plexuses while arachnoidal arteries were poorly innervated by only single nerves. The vascular nerve fibers formed bundles varying in thickness, from very thick bundles often interchanging nerve fibers to quite thin fascicles. Thick bundles were absent from the walls of medium sized vessels and small meningeal arteries where only smaller fascicles or single fibres occurred. NADPH-d-positive neurons and nerve fibres were found in all the ganglia investigated. However, pronounced differences in the number of the positive nerve structures were observed between the ganglia. In the pterygopalatine, trigeminal and sensory ganglia of the vagal nerve the vast majority of neurons were NADPH-d-positive. Numerous NADPH-d-positive nerve fibers occurred within the pterygopalatine and trigeminal ganglion while sensory ganglia of the vagal nerve comprised smaller number of fibres. Small numbers of the neurons and moderate numbers of the nerve fibres occurred in the cranial cervical ganglion. The intensity of NADPH-d reaction in the endothelium was constant independent of the size of the vessels studied.
Animals↗
Occurrence and coexistence of some neuropeptides in nerve fibers supplying the bovine ovary and its extrinsic blood vessels.
Single- and double-labelling immunofluorescence was applied to investigate the occurrence and coexistence of vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) and substance P (SP) in nerve fibers supplying the bovine ovary as well as its extrinsic blood vessels. We also studied the coexistence of these neuropeptides with tyrosine hydroxylase (TH) and neuropeptide Y (NPY) to determine features of "chemical coding" of nerve fibers innervating this organ. The bovine ovary was found to be supplied by VIP- and SP-immunoreactive (VIP- and SP-IR) nerve fibers. No intraovarian CGRP-IR nerve terminals were detected. Immunoreactivity to VIP was shown in a moderate number of nerve fibers occurring in all ovarian regions - ovarian hilus, cortex and medulla. VIP-IR nerves were found to innervate blood vessels and ovarian follicles, especially primordial and primary ones. Immunoreactivity to SP was detected only in solitary intraovarian nerve fibers which were also encountered in all ovarian regions, but with special preference to ovarian medulla, where they innervated blood vessels. SP-IR nerve fibres were sometimes observed in close vicinity to primordial and primary follicles. Double-labelling immunofluorescence revealed that some VIP-IR nerve fibres supplying intraovarian blood vessels were simultaneously TH- or NPY-IR. Small numbers of VIP-IR fibres contained immunoreactivity to SP. All SP-IR nerve terminals were TH-or NPY-negative. The ovarian artery as well as arteries of the paraovarian plexus were moderately supplied with VIP-, or by single CGRP- or SP-IR nerve fibres. Veins were innervated by only solitary nerve terminals containing these neuropeptides. The following patterns of coexistence were determined in nerve fibres supplying the investigated blood vessels: VIP, VIP/CGRP, VIP/SP, CGRP/SP, VIP/NPY. No distinct differences in the innervation of the ovary and its extrinsic blood vessels between juvenile and adult cows were found.
Age Factors↗
Immunohistochemical study of the existence and coexistence of catecholamine synthesizing enzymes and some neuropeptides in perivascular nerve fibres of the main thoraco-cranial arteries in the pig.
The presence and pattern of coexistence of catecholamine-synthesizing enzymes and some neuropeptides in nerve fibres supplying thoraco-cranial arteries of the sexually immature gilts were investigated in whole mount preparations. The studied substances included: tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (D beta H) (as markers of catecholaminergic nerve fibres), neuropeptide Y (NPY), Leu5-enkephalin (LENK), vasoactive intestinal polipeptide (VIP), calcitonin gene-related peptide (CGRP), substance P (SP), galanin (GAL), somatostatin (SOM) and serotonin (5-HT). The arteries were found to be richly supplied by TH/D beta H-immunoreactive (TH/D beta H) nerve fibers. Of the neuropeptides studied, NPY (rich innervation), LENK (moderate innervation), VIP (moderate innervation) and CGRP, SP, GAL (only a few nerve fibres) were detected in periarterial nerves. The following patterns of coexistence of the studied substances were found: TH+/D beta H+, TH+/D beta H+/NPY+, TH+/D beta H+/LENK+, TH-/D beta H-/NPY+, TH-/D beta H-/VIP+, TH+/D beta H+/VIP+ (only a few nerve fibres in the cerebral arteries), LENK+/NPY+, LENK-/NPY+, LENK+/NPY-. No SOM and 5-HT-positive structures were observed in the porcine blood vessels.
Animals↗
Immunohistochemical localization of neuropeptides in the porcine thoraco-lumbar paravertebral ganglia.
The existence and distribution pattern of neuropeptide Y, Met5-enkephalin-Arg6-Gly7-Leu8, vasoactive intestinal polypeptide, calcitonin gene-related peptide, substance P and bombesin/gastrin releasing peptide in the neuronal elements of the thoracolumbar paravertebral ganglia (T4-L6) were studied immunohistochemically in sexually immature female pigs. Subpopulations of nerve cell bodies containing immunoreactivity to neuropeptide Y, Met5-enkephalin-Arg6-Gly7-Leu8, vasoactive intestinal polypeptide and calcitonin gene-related peptide were described. However, neurons were non-immunoreactive for substance P and bombesin/gastrin releasing peptide. The solitary small intensely fluorescent cells contain calcitonin gene-related peptide-, substance P- and Met5-enkephalin-Arg6-Gly7-Leu8-, whereas the some cells in clusters contained only substance P and only substance P and Met5-enkephalin-Arg6-Gly7-Leu8. Immunoreactivities to all studied peptides occurred in the nerve fibres within investigated ganglia. Additionally the number of nerve fibres containing particular peptides as well as their distribution pattern were found to vary. The results of this study were compared with those of previous investigations in other species.
Amino Acid Sequence↗
NO-synthase-containing neurons of the pig inferior mesenteric ganglion, part of them innervating the ductus deferens.
The presence of nitric oxide-synthase (NOS) in neurons of the porcine inferior mesenteric ganglion (IMG) has been investigated. A minority (about 1-3%) of the neurons were immunoreactive (IR) for NOS, the vast majority of which stained for neuropeptide Y (NPY) but not for tyrosine hydroxylase (TH). A small subpopulation of prevertebral neurons, 1% of which stained for NOS or NADPH-diaphorase (NADPHd), projected to the ductus deferens, as demonstrated by retrograde tracing. Within the wall of the ductus deferens, NOS- or NOS/NPY-IR nerve fibres were found to innervate the smooth muscle or were closely associated with blood vessels. It is therefore suggested that nitric oxide might be involved in the regulation of local blood flow and muscular tone in the wall of the pig ductus deferens.
Amino Acid Oxidoreductases↗
Immunohistochemical localization of neuropeptides in nerve fibers of the porcine vagina and uterine cervix.
The distribution of vasoactive intestinal polypeptide, substance P, Met5-enkephalin-Arg6-Gly7-Leu8, neuropeptide Y, calcitonin gene-related peptide and bombesin/gastrin releasing peptide was studied immunohistochemically in nerve fibres supplying the vagina and uterine cervix of sexually immature pigs. Nerves containing neuropeptide Y- and Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactivity were particularly numerous in the uterine cervix and vagina. Vasoactive intestinal polypeptide- and and calcitonin gene-related peptide-containing nerves were less numerous, whereas the bombesin/gastrin releasing peptide- and substance P-immunoreactive nerves were scarce in these organs. Generally, the immunoreactive fibres, nerve networks or bundles were distributed under the serous membrane, among smooth muscle of muscular membranes, in the submucosal layer and under the luminar epithelium of the uterine cervix and vagina.
Amino Acid Sequence↗
Immunohistochemical colocalization of Met5-enkephalin-Arg6-Gly7-Leu8 with tyrosine hydroxylase in neurons of the porcine inferior mesenteric and coeliac-superior mesenteric ganglia.
The localization and distribution of the proenkephalin A-derived octapeptide Met5-enkephalin-Arg6-Gly7-Leu8 (MEAGL) was studied in the porcine sympathetic prevertebral ganglia by using the indirect immunofluorescence technique. In the inferior mesenteric ganglion, a small population of principal neurons and SIF cells contained intense MEAGL immunoreactivity. A very dense plexus of MEAGL-immunoreactive nerve fibers was distributed in different parts of the ganglion. A low number of MEAGL- immunoreactive principal ganglion cells and SIF cells, as well as numerous nerve fibers were also present in the coeliac-superior mesenteric ganglion. Double staining experiments indicated that the MEAGL-immunoreactive principal neurons in the inferior mesenteric and coeliac-superior mesenteric ganglia also contained tyrosine hydroxylase immunoreactivity.
Animals↗
Immunocytochemical demonstration of neuropeptide Y and luteinizing hormone-releasing hormone-immunoreactive structures in the organum vasculosum laminae terminalis of juvenile gilts.
Immunocytochemical investigations on the immature gilt organum vasculosum laminae terminalis showed extensive neuropeptide Y- and luteinizing hormone-releasing hormone-immunoreactive innervation of the organ. The luteinizing hormone-releasing hormone-containing varicose fibers ran along the organum vasculosum laminae terminalis in close association with blood vessels. The nerve processes originating from well-stained luteinizing hormone-releasing hormone-immunoreactive perikarya were distributed around the organum vasculosum laminae terminalis. The matrix of the organum vasculosum laminae terminalis was abundantly supplied by neuropeptide Y-immunoreactive varicose fibers. Numerous neuropeptide Y-immunoreactive terminals seemed to penetrate the ependymal lining of the organ. From these observations, it is concluded that there are favorable morphological conditions for secretion of neuropeptide Y into the cerebrospinal fluid of the third ventricle and release of luteinizing hormone-releasing hormone into fenestrated capillaries of the organ.
Animals↗
Distribution of neuropeptides in the porcine stellate ganglion.
The localization and distribution of neuropeptides including neuropeptide Y (NPY), [Met5]enkephalin-Arg6-Gly7-Leu8 (MEAGL), vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), substance P and somatostatin (SOM) were analyzed in the stellate ganglion of the pig by use of the indirect immunofluorescence technique. NPY, MEAGL, SOM, VIP and CGRP immunoreactivities were found to exist in subpopulations of neuronal cell bodies of the stellate ganglion. A population of the small intensely fluorescent (SIF) cells showed MEAGL immunoreactivity. In addition, the presence of NPY-, MEAGL-, CGRP-, SP-, SOM- and VIP-immunoreactive nerve fibers and axonal varicosities were observed in the stellate ganglion. The localization and pattern of distribution of these peptides in the porcine stellate ganglion were compared with studies carried out on stellate ganglia of other mammalian species.
Animals↗
Neuropeptides in the porcine coeliac-superior mesenteric ganglion.
The occurrence and distribution of tyrosine hydroxylase, neuropeptide Y, somatostatin, Met5-enkephalin-Arg6-Gly7-Leu8, vasoactive intestinal polypeptide, substance P, calcitonin gene-related peptide, bombesin gastrin releasing peptide and galanin were immunohistochemically studied in the perikarya and nerve fibres of the porcine coeliac-superior mesenteric ganglion of untreated juvenile pigs. Subpopulations of neurons containing immunoreactivities to tyrosine hydroxylase, neuropeptide Y, Met5-enkephalin-Arg6-Gly7-Leu8, somatostatin, vasoactive intestinal polypeptide and galanin were disclosed in the studied ganglion, whereas principal ganglionic cells were non-immunoreactive for other investigated peptides. Double-immunofluorescence and analysis of consecutive sections revealed a partial colocalization of tyrosine hydroxylase and neuropeptide Y, Met5-enkephalin-Arg6-Gly7-Leu8 and somatostatin, whereas immunoreactivity to vasoactive intestinal polypeptide and/or to neuropeptide Y was found in non-noradrenergic neurons in this ganglion. All of neuropeptides studied were found in nerve fibres in this ganglion. The results of this study were compared with those of previous studies performed on other species.
Amino Acid Sequence↗