Waldemar Sienkiewicz - Profile on Academia.edu (original) (raw)

Papers by Waldemar Sienkiewicz

Neuropeptides and catecholamines are biologically active substances which play the roles of neuro... more Neuropeptides and catecholamines are biologically active substances which play the roles of neurotransmitters, neuromodulators and cotransmitters in the central nervous system (CNS). These substances are known to regulate and influence a wide spectrum of functions; such as food and water intake, thermoregulation, growth and maturation, sexual behaviour, reproduction, function of the hypothalamic-hypophysial-organ axes, and many others. This large family of neuropeptides is comprised of around 40 fundamental polypeptides, which does not include their derivatives and related peptides. Localization and development of several of them was intensively studied in the brain of a wide range of species, including the rat and the pig. In this present paper, data regarding distribution of catecholamine synthesising enzymes and some neuropeptides, as well as their gene expression during development in the rodent and porcine brain are summarised and related to development of LHRH-containing structures.

Immunohistochemical Properties of Caudal Mesenteric Ganglion and Anterior Pelvic Ganglion Neurons Projecting to the Porcine Testes in Animals Subjected to Hemicastration, Castration, and Testosterone Supplementation

Bulletin of The Veterinary Institute in Pulawy, 2010

ABSTRACT The present study was carried out on sexually mature boars. All the animals were injecte... more ABSTRACT The present study was carried out on sexually mature boars. All the animals were injected with fast blue into right testis and then divided into four groups (G1- control animals, G2 - hemicastrated, G3 – castrated, and G4 - castrated and injected with testosterone). After 3 weeks, G1 pigs were transcardially perfused. In G2 pigs right testes, whereas in G3 and G4 animals both testes were removed. G4 pigs were injected with testosterone. After 2 weeks, the pigs were transcardially perfused and then their caudal mesenteric ganglia (CaMG) and anterior pelvic ganglia (APG) were collected. The ganglia were cut into 12 µm-thick cryostat sections. Sections were stained using antisera against TH or DßH, VACHT or CHAT, NPY, VIP and GAL, and androgen receptor (AR). Immunohistochemical staining of CaMG-sections revealed that approximately 74% of FB-positive (FB+ ) neurons contained immunoreactivity to TH or DßH, whereas 4% of FB+ cells were VACHT-positive. Among FB+ /DßH+ neurons, 72% contained NPY and 2% stained for GAL. All FB+ /VACHT+ neurons were also VIP+ . 62% of FB+ somata were NPY+ , whereas 6% stained for VIP. In all experimental animals, numbers of FB+ perikarya immunoreactive to TH (approx. 30%) and DßH (approx. 50%) were smaller than those found in G1 animals, whereas numbers of neurons displaying immunoreactivity to other substances studied were higher. The most significant increases regarded those expressing GAL (approx. 30%) and VIP (approx. 20%) whereas less distinct changes dealt with NPY+ and VAChT+ or ChAT+ neurones. In APG, 60% of FB+ neurons contained immunoreactivity to TH or DßH, whereas 12% of FB+ cells were VACHT-positive. Among FB+ /DßH+ neurons, 55% contained NPY and 3% stained for GAL. All FB+ /VACHT+ neurons were also VIP+ . 46% of FB+ somata were NPY-IR, whereas 19% stained for VIP. In all experimental pigs, the immunohistochemical properties of the APG FB+ neurones were similar to those found in relation to CaMG-perikarya. Neurones in of both studied ganglia were surrounded by dense networks of VACHT-positive nerve fibres. The most apparent changes in the immunohistochemical features of the FB+ neurons evoked by bilateral castration were observed in G3 pigs; whereas changes found in G4 were very similar to those observed in G2 animals.

Ontogeny of the expression of some catecholamine synthesising enzymes in the female porcine preoptic area

PubMed, 2002

Ontogeny of the catecholaminergic system of the preoptic area (PA) was studied in various animal ... more Ontogeny of the catecholaminergic system of the preoptic area (PA) was studied in various animal species including mice, rats, cats and lower vertebrates. Until now, there has been no data about development of catecholaminergic structures in the porcine PA. To study this problem, hypothalami from six groups of animals were collected. Three groups of foetuses (70, 84 and 112 days old) and three groups of female pigs (1 day, 10 weeks and 7-8 months old) were used. Nerve structures immunoreactive for the studied substances: tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DbetaH) and phenylethanoloamine-N-metylthransferase (PNMT) were observed in different periods. In PA, TH-IR (immunoreactive) structures appeared before 70th day of foetal life, DbetaH-IR between 70th and 84th day of foetal live and PNMT-IR only in 10-week old and adult animals. In the PA of 70-day old foetuses, single smooth and varicose nerve fibres immunoreactive only to TH were found. In PA of 84-day old foetuses, additionally, single nerve cell bodies immunoreactive to TH were shown and some of them also contained immunoreactivity to DbetaH. In PA of 1-day old piglets, moderate numbers of nerve fibres immunoreactive to TH and only single TH/DbetaH-IR nerve terminals were observed. TH-IR nerve cell bodies were also moderate in number and many of them contained simultaneously immunoreactivity to DbetaH. In PA of 10-week old pigs, a moderate number of immunopositive nerve fibres was observed. They contained mainly TH, but part of them stained also for TH/DbetaH. Only very few nerve fibres containing exclusively DbetaH were found. These nerve terminals were observed in a close vicinity of blood vessels. In PA, moderate numbers of TH-IR nerve cell bodies were found, some of them contained also immunoreactivity to DH but never to PNMT. Perikarya containing PNMT were TH-negative. In the PA of sexually mature sows, additional, single, large nerve cell bodies (about 35 microm in a diameter) containing TH only were found. In many cases, TH- and DbetaH-IR "basket-like" structures surrounding nerve cell bodies were seen, suggesting an influence of those fibres on the neuronal activity.

Polish Journal of Veterinary Sciences, 2009

This is the first report dealing with the localisation and morphology of the proximal (jugular) g... more This is the first report dealing with the localisation and morphology of the proximal (jugular) ganglion in the pig. Six 3-months-old pigs of both sexes were used in this study. Tissues were stained with three histological methods: Klüver-Barer counterstained with Cresyl violet, Haematoxylin-eosin and Mayer's haematoxylin. The localisation and morphological features of the ganglion and ganglionic neurones were described and discussed.

Ontogeny of some katecholamines synthetising enzymes in some nuclei of female pigs hypothalamus

Folia Morphologica, 1998

Immunocastration-induced changes in the pattern of the porcine seminal vesicle innervation

Occurrence and coexistence of some neuropeptides in nerve fibers supplying the bovine ovary and its extrinsic blood vessels

PubMed, 1995

Single- and double-labelling immunofluorescence was applied to investigate the occurrence and coe... more 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.

Biology, Jan 30, 2021

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

The Distribution and Chemical Coding of Intramural Neurons Supplying the Porcine Stomach ? the Study on Normal Pigs and on Animals Suffering from Swine Dysentery

Anatomia Histologia Embryologia, Jun 1, 2007

SummaryThe present study was designed to investigate the expression of biologically active substa... more SummaryThe present study was designed to investigate the expression of biologically active substances by intramural neurons supplying the stomach in normal (control) pigs and in pigs suffering from dysentery. Eight juvenile female pigs were used. Both dysenteric (n = 4; inoculated with Brachyspira hyodysenteriae) and control (n = 4) animals were deeply anaesthetized, transcardially perfused with buffered paraformalehyde, and tissue samples comprising all layers of the wall of the ventricular fundus were collected. The cryostat sections were processed for double‐labelling immunofluorescence to study the distribution of the intramural nerve structures (visualized with antibodies against protein gene‐product 9.5) and their chemical coding using antibodies against vesicular acetylcholine (ACh) transporter (VAChT), nitric oxide synthase (NOS), galanin (GAL), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), Leu5‐enkephalin (LENK), substance P (SP) and calcitonin gene‐related peptide (CGRP). In both inner and outer submucosal plexuses of the control pigs, the majority of neurons were SP (55% and 58%, respectively)‐ or VAChT (54%)‐positive. Many neurons stained also for CGRP (43 and 45%) or GAL (20% and 18%) and solitary perikarya were NOS‐, SOM‐ or VIP‐positive. The myenteric plexus neurons stained for NOS (20%), VAChT (15%), GAL (10%), VIP (7%), SP (6%) or CGRP (solitary neurons), but they were SOM‐negative. No intramural neurons immunoreactive to LENK were found. The most remarkable difference in the chemical coding of enteric neurons between the control and dysenteric pigs was a very increased number of GAL‐ and VAChT‐positive nerve cells (up to 61% and 85%, respectively) in submucosal plexuses of the infected animals. The present results suggest that GAL and ACh have a specific role in local neural circuits of the inflamed porcine stomach in the course of swine dysentery.

Veterinarni Medicina, May 31, 2018

Inflammatory processes are associated with changes in the interplay of different pro-and antiinfl... more Inflammatory processes are associated with changes in the interplay of different pro-and antiinflammatory factors, including neuropeptides, in tissue. This study was performed to investigate the influence of proliferative enteropathy on the concentration of several neuropeptides known to be involved in the regulation of the inflammatory process in porcine intestine and intestine-innervating ganglia. The concentration of galanin, vasoactive intestinal polypeptide, somatostatin, neuropeptide Y, substance P and calcitonin gene-related peptide were assayed with ELISA in the coeliac-superior mesenteric ganglion, inferior mesenteric ganglion, selected dorsal root ganglia, ileum and the descending colon in healthy and sick pigs. The concentrations of the studied neuropeptides were higher in sick animals. Statistically significant differences were found for coeliac-superior mesenteric ganglion (galanin, vasoactive intestinal polypeptide, somatostatin and neuropeptide Y), inferior mesenteric ganglion (galanin, somatostatin and neuropeptide Y), dorsal root ganglia (galanin, somatostatin, neuropeptide Y and calcitonin gene-related peptide), ileum (galanin and somatostatin) and the descending colon (galanin, somatostatin and neuropeptide Y). The data clearly show the influence of the inflammatory process on the concentration of some of the studied neuropeptides present in inflamed tissues and ganglia innervating the inflamed gut. These changes must be associated with the role the studied neuropeptides play in the inflammatory process.

Acta Zoologica, Sep 11, 2014

Dudek, A., Sienkiewicz, W. and Kaleczyc, J. 2014. Contribution of the dorsal branch of the access... more Dudek, A., Sienkiewicz, W. and Kaleczyc, J. 2014. Contribution of the dorsal branch of the accessory nerve to the innervation of the trapezius muscle in the piga retrograde tracing study-Acta Zoologica (Stockholm) 00: 000-000. The study was aimed to establish the contribution of the dorsal branch of the accessory nerve (DBXI) to the innervation of the porcine trapezius muscle (TRAP). Combined retrograde tracing using fluorescent tracer Fast blue (FB) and surgical denervation procedure (excision of DBXI segment) were applied. FB+ neurons supplying the cervical (c-TRAP) and thoracic part (th-TRAP) of TRAP were localized in following nerve centres: the ipsilateral ventral horn of the grey matter of cervical neuromers, ipsilateral spinal ganglia and bilateral sympathetic chain ganglia. After the excision of DBXI segment, no FB+ motoneurons supplying c-TRAP were found while the mean number of those supplying th-TRAP was significantly decreased. A slight decrease in average numbers of sensory and autonomic neurons implemented in the innervation of both parts of TRAP was also observed. This study has revealed that in the pig DBXI is the only motor pathway to c-TRAP while the vast majority of motoneurons supplying porcine th-TRAP send their axons via DBXI.

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

PubMed, 1995

The presence and pattern of coexistence of catecholamine-synthesizing enzymes and some neuropepti... more 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.

Existence and coexistence of catecholamine-synthesizing enzymes and some neuropeptides in nerve fibres supplying the bovine ovary and its blood vessels

The preoptic area in the sexually immature pig. Morphological study

PubMed, 1993

The preoptic area was investigated in 5 immature pigs. This area is composed of three nuclei: lat... more The preoptic area was investigated in 5 immature pigs. This area is composed of three nuclei: lateral, medial, and median. The largest is the lateral nucleus.

Ontogeny of the expression of catecholamine synthesising enzymes in the female porcine median eminence arcuate nucleus complex (MEARC)

PubMed, 2003

The ontogeny of the catecholaminergic system of the median eminence (ME) arcuate nucleus (ARC) co... more The ontogeny of the catecholaminergic system of the median eminence (ME) arcuate nucleus (ARC) complex (MEARC) has been studied in various animal species but so far, nothing has been learnt about the development of catecholaminergic structures in the porcine MEARC. To study this problem the hypothalami from animals at different ages (six groups) were collected. Nerve structures immunoreactive (R) for the substances studied [(tyrosine hydroylase (TH), dopamine beta-hydroxylase (D(beta)H) and phenylethanoloamine-N-metylthransferase (PNMT)] were found in the pigs at different age periods. In MEARC, TH-IR structures appeared before the 70th day of foetal life, D(beta)H-IR before the 10th week of postnatal life and PNMT-IR only in sexually mature sows.

Distribution of neuropeptide Y-like immunoreactivity in the hypothalamic tuberal nuclei of the immature gilts

PubMed, 1994

The distribution of neuropeptide Y in the immature female pig hypothalamic tuberal nuclei was inv... more The distribution of neuropeptide Y in the immature female pig hypothalamic tuberal nuclei was investigated using avidin-biotin complex method. This study showed cell bodies containing NPY-immunoreactive material in the arcuate nucleus, dorsomedial and ventromedial nuclei, as well as internal layer of the median eminence and infundibulum, an observation not yet reported in the pig. NPY-immunoreactive fibers were noted throughout the hypothalamus, but most prominently within the periventricular region. Median eminence disclosed abundance of NPY-immunoreactive processes. The study shows a detailed organization of cell bodies and fibers containing NPY-immunoreactivity in tuberal hypothalamic nuclei of the immature gilts.

Immunohistochemical properties of sympathetic chain ganglia (SChG) neurons projecting to the porcine testis in animals subjected to hemicastration, castration and testosterone supplementation

PubMed, 2010

The present study was carried out on sexually mature boars. All the animals were injected with Fa... more The present study was carried out on sexually mature boars. All the animals were injected with Fast Blue into the right testis and then divided into four groups [(group 1--control (G1), group 2--hemicastreatad (G2), group 3 castrated (G3) and group 4--castrated and injected with testosterone (G4) boars)]. After a survival period of 3 weeks, G1 animals were transcardially perfused. In pigs of G2, right testis was removed, whereas in G3 and G4 animals both testes were removed. The pigs of G4 were injected with testosterone. After two weeks, all the animals were transcardially perfused and then their sympathetic chain ganglia were collected. The ganglia were cut into 12 microm-thick cryostat sections. The sections were stained using antisera against TH or DbetaH, VACHT or CHAT, NPY, VIP, GAL and AR. Testosterone plasma levels were evaluated with ELISA test. In control pigs, testosterone level amounted to 8.51 +/- 1.29 ng/ml, whereas in hemicastrated animals it was 1.72 +/- 0.35 ng/ml. Bilateral orchiectomy resulted in a drastic decline in testosterone level. Administration of exogenous hormone resulted in tremendous increase in its plasma level. In control pigs, FB-positive (FB+) neurons were found in the right and left ganglia. Immunohistochemical staining revealed that approximately 74% of FB+ neurons contained immunoreactivity to TH or DbetaH, whereas approximately 4% of FB+ cells were VACHT-positive. Among FB+/DbetaH+ neurons, 72% contained NPY and less than 1% stained for GAL. All FB+/VACHT+ neurons were also VIP+. Approximately 62% of FB+ somata expressed immunoreactivity to NPY, whereas 6% stained for VIP. In all experimental pigs, numbers of FB+ perikarya immunoreactive to TH (from 21% in G3, 26% in G2 to 29% in G4,) and DbetaH (62% in G2, 58% in G3 and 59% in G4,) were smaller than those found in G1 animals, whereas numbers of neurons displaying immunoreactivity to other substances studied were higher. The most significant increases in the number of immunoreactive cells were observed in regard to those expressing GAL (24% in G2, 30% in G3 and 27% in G4,) and VIP (11% in G2, 12% in G3 and 11% in G4,) whereas less distinct changes were observed in case of neurones which stained to NPY (67% in G2, 70% in G3 and 68% in G4,) and VAChT or ChAT (6% in G2, G3 and G4). The most apparent changes in the immunohistochemical features of FB+ neurons evoked by bilateral castration were observed in G3 pigs, whereas changes found in G4 animals were very similar to those observed in G2 pigs.

NADPH-diaphorase-containing cerebrovascular nerve fibres and their possible origin in the pig

PubMed, 1995

NADPH-diaphorase histochemical technique was applied to demonstrate the catalytic activity of nit... more 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.

Changes in the expression of some neuropeptides in the intestines and nerve ganglia during the porcine dysentery

Bulletin of The Veterinary Institute in Pulawy, 2005

Histochemistry and Cell Biology, 2020

The present study investigated the influence of castration performed at neonatal age on neuronal ... more The present study investigated the influence of castration performed at neonatal age on neuronal elements in the anterior pelvic ganglion of the male pig with immunohistochemistry and quantitative real-time PCR (qPCR). The ganglia were examined 3 and 6 months after surgery. In 3-month-old castrated pigs (3MCP) 74% of adrenergic and 31% of cholinergic neurons stained for caspase-3 (CASP-3), and much greater numbers of perikarya than in the control animals expressed CGRP, galanin (GAL) and VIP (peptides known to have neuroprotective properties). In 6-months-old castrated pigs (6MCP), an excessive loss (90%) of neurons and intraganglionic nerve fibres was found. The survived adrenergic and cholinergic neurons also expressed CASP-3, CGRP, GAL or VIP. The qPCR results corresponded with immunofluorescence findings. In 3MCP, genes for CASP-3 and CGRP were up-regulated, while the expression of those for DβH, VAChT, GAL, VIP and SP displayed statistically insignificant variations. In 6MCP, distinctly up-regulated were genes for CGRP, GAL, VIP, SP, DβH and VAChT, while the expression of casp3 gene was down-regulated. The study revealed for the first time the excessive loss of pelvic neurons following castration, and a realistic assumption is proposed, that the neurons died due to apoptosis triggered by androgen deprivation.

Neuropeptides and catecholamines are biologically active substances which play the roles of neuro... more Neuropeptides and catecholamines are biologically active substances which play the roles of neurotransmitters, neuromodulators and cotransmitters in the central nervous system (CNS). These substances are known to regulate and influence a wide spectrum of functions; such as food and water intake, thermoregulation, growth and maturation, sexual behaviour, reproduction, function of the hypothalamic-hypophysial-organ axes, and many others. This large family of neuropeptides is comprised of around 40 fundamental polypeptides, which does not include their derivatives and related peptides. Localization and development of several of them was intensively studied in the brain of a wide range of species, including the rat and the pig. In this present paper, data regarding distribution of catecholamine synthesising enzymes and some neuropeptides, as well as their gene expression during development in the rodent and porcine brain are summarised and related to development of LHRH-containing structures.

Immunohistochemical Properties of Caudal Mesenteric Ganglion and Anterior Pelvic Ganglion Neurons Projecting to the Porcine Testes in Animals Subjected to Hemicastration, Castration, and Testosterone Supplementation

Bulletin of The Veterinary Institute in Pulawy, 2010

ABSTRACT The present study was carried out on sexually mature boars. All the animals were injecte... more ABSTRACT The present study was carried out on sexually mature boars. All the animals were injected with fast blue into right testis and then divided into four groups (G1- control animals, G2 - hemicastrated, G3 – castrated, and G4 - castrated and injected with testosterone). After 3 weeks, G1 pigs were transcardially perfused. In G2 pigs right testes, whereas in G3 and G4 animals both testes were removed. G4 pigs were injected with testosterone. After 2 weeks, the pigs were transcardially perfused and then their caudal mesenteric ganglia (CaMG) and anterior pelvic ganglia (APG) were collected. The ganglia were cut into 12 µm-thick cryostat sections. Sections were stained using antisera against TH or DßH, VACHT or CHAT, NPY, VIP and GAL, and androgen receptor (AR). Immunohistochemical staining of CaMG-sections revealed that approximately 74% of FB-positive (FB+ ) neurons contained immunoreactivity to TH or DßH, whereas 4% of FB+ cells were VACHT-positive. Among FB+ /DßH+ neurons, 72% contained NPY and 2% stained for GAL. All FB+ /VACHT+ neurons were also VIP+ . 62% of FB+ somata were NPY+ , whereas 6% stained for VIP. In all experimental animals, numbers of FB+ perikarya immunoreactive to TH (approx. 30%) and DßH (approx. 50%) were smaller than those found in G1 animals, whereas numbers of neurons displaying immunoreactivity to other substances studied were higher. The most significant increases regarded those expressing GAL (approx. 30%) and VIP (approx. 20%) whereas less distinct changes dealt with NPY+ and VAChT+ or ChAT+ neurones. In APG, 60% of FB+ neurons contained immunoreactivity to TH or DßH, whereas 12% of FB+ cells were VACHT-positive. Among FB+ /DßH+ neurons, 55% contained NPY and 3% stained for GAL. All FB+ /VACHT+ neurons were also VIP+ . 46% of FB+ somata were NPY-IR, whereas 19% stained for VIP. In all experimental pigs, the immunohistochemical properties of the APG FB+ neurones were similar to those found in relation to CaMG-perikarya. Neurones in of both studied ganglia were surrounded by dense networks of VACHT-positive nerve fibres. The most apparent changes in the immunohistochemical features of the FB+ neurons evoked by bilateral castration were observed in G3 pigs; whereas changes found in G4 were very similar to those observed in G2 animals.

Ontogeny of the expression of some catecholamine synthesising enzymes in the female porcine preoptic area

PubMed, 2002

Ontogeny of the catecholaminergic system of the preoptic area (PA) was studied in various animal ... more Ontogeny of the catecholaminergic system of the preoptic area (PA) was studied in various animal species including mice, rats, cats and lower vertebrates. Until now, there has been no data about development of catecholaminergic structures in the porcine PA. To study this problem, hypothalami from six groups of animals were collected. Three groups of foetuses (70, 84 and 112 days old) and three groups of female pigs (1 day, 10 weeks and 7-8 months old) were used. Nerve structures immunoreactive for the studied substances: tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DbetaH) and phenylethanoloamine-N-metylthransferase (PNMT) were observed in different periods. In PA, TH-IR (immunoreactive) structures appeared before 70th day of foetal life, DbetaH-IR between 70th and 84th day of foetal live and PNMT-IR only in 10-week old and adult animals. In the PA of 70-day old foetuses, single smooth and varicose nerve fibres immunoreactive only to TH were found. In PA of 84-day old foetuses, additionally, single nerve cell bodies immunoreactive to TH were shown and some of them also contained immunoreactivity to DbetaH. In PA of 1-day old piglets, moderate numbers of nerve fibres immunoreactive to TH and only single TH/DbetaH-IR nerve terminals were observed. TH-IR nerve cell bodies were also moderate in number and many of them contained simultaneously immunoreactivity to DbetaH. In PA of 10-week old pigs, a moderate number of immunopositive nerve fibres was observed. They contained mainly TH, but part of them stained also for TH/DbetaH. Only very few nerve fibres containing exclusively DbetaH were found. These nerve terminals were observed in a close vicinity of blood vessels. In PA, moderate numbers of TH-IR nerve cell bodies were found, some of them contained also immunoreactivity to DH but never to PNMT. Perikarya containing PNMT were TH-negative. In the PA of sexually mature sows, additional, single, large nerve cell bodies (about 35 microm in a diameter) containing TH only were found. In many cases, TH- and DbetaH-IR "basket-like" structures surrounding nerve cell bodies were seen, suggesting an influence of those fibres on the neuronal activity.

Polish Journal of Veterinary Sciences, 2009

This is the first report dealing with the localisation and morphology of the proximal (jugular) g... more This is the first report dealing with the localisation and morphology of the proximal (jugular) ganglion in the pig. Six 3-months-old pigs of both sexes were used in this study. Tissues were stained with three histological methods: Klüver-Barer counterstained with Cresyl violet, Haematoxylin-eosin and Mayer's haematoxylin. The localisation and morphological features of the ganglion and ganglionic neurones were described and discussed.

Ontogeny of some katecholamines synthetising enzymes in some nuclei of female pigs hypothalamus

Folia Morphologica, 1998

Immunocastration-induced changes in the pattern of the porcine seminal vesicle innervation

Occurrence and coexistence of some neuropeptides in nerve fibers supplying the bovine ovary and its extrinsic blood vessels

PubMed, 1995

Single- and double-labelling immunofluorescence was applied to investigate the occurrence and coe... more 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.

Biology, Jan 30, 2021

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

The Distribution and Chemical Coding of Intramural Neurons Supplying the Porcine Stomach ? the Study on Normal Pigs and on Animals Suffering from Swine Dysentery

Anatomia Histologia Embryologia, Jun 1, 2007

SummaryThe present study was designed to investigate the expression of biologically active substa... more SummaryThe present study was designed to investigate the expression of biologically active substances by intramural neurons supplying the stomach in normal (control) pigs and in pigs suffering from dysentery. Eight juvenile female pigs were used. Both dysenteric (n = 4; inoculated with Brachyspira hyodysenteriae) and control (n = 4) animals were deeply anaesthetized, transcardially perfused with buffered paraformalehyde, and tissue samples comprising all layers of the wall of the ventricular fundus were collected. The cryostat sections were processed for double‐labelling immunofluorescence to study the distribution of the intramural nerve structures (visualized with antibodies against protein gene‐product 9.5) and their chemical coding using antibodies against vesicular acetylcholine (ACh) transporter (VAChT), nitric oxide synthase (NOS), galanin (GAL), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), Leu5‐enkephalin (LENK), substance P (SP) and calcitonin gene‐related peptide (CGRP). In both inner and outer submucosal plexuses of the control pigs, the majority of neurons were SP (55% and 58%, respectively)‐ or VAChT (54%)‐positive. Many neurons stained also for CGRP (43 and 45%) or GAL (20% and 18%) and solitary perikarya were NOS‐, SOM‐ or VIP‐positive. The myenteric plexus neurons stained for NOS (20%), VAChT (15%), GAL (10%), VIP (7%), SP (6%) or CGRP (solitary neurons), but they were SOM‐negative. No intramural neurons immunoreactive to LENK were found. The most remarkable difference in the chemical coding of enteric neurons between the control and dysenteric pigs was a very increased number of GAL‐ and VAChT‐positive nerve cells (up to 61% and 85%, respectively) in submucosal plexuses of the infected animals. The present results suggest that GAL and ACh have a specific role in local neural circuits of the inflamed porcine stomach in the course of swine dysentery.

Veterinarni Medicina, May 31, 2018

Inflammatory processes are associated with changes in the interplay of different pro-and antiinfl... more Inflammatory processes are associated with changes in the interplay of different pro-and antiinflammatory factors, including neuropeptides, in tissue. This study was performed to investigate the influence of proliferative enteropathy on the concentration of several neuropeptides known to be involved in the regulation of the inflammatory process in porcine intestine and intestine-innervating ganglia. The concentration of galanin, vasoactive intestinal polypeptide, somatostatin, neuropeptide Y, substance P and calcitonin gene-related peptide were assayed with ELISA in the coeliac-superior mesenteric ganglion, inferior mesenteric ganglion, selected dorsal root ganglia, ileum and the descending colon in healthy and sick pigs. The concentrations of the studied neuropeptides were higher in sick animals. Statistically significant differences were found for coeliac-superior mesenteric ganglion (galanin, vasoactive intestinal polypeptide, somatostatin and neuropeptide Y), inferior mesenteric ganglion (galanin, somatostatin and neuropeptide Y), dorsal root ganglia (galanin, somatostatin, neuropeptide Y and calcitonin gene-related peptide), ileum (galanin and somatostatin) and the descending colon (galanin, somatostatin and neuropeptide Y). The data clearly show the influence of the inflammatory process on the concentration of some of the studied neuropeptides present in inflamed tissues and ganglia innervating the inflamed gut. These changes must be associated with the role the studied neuropeptides play in the inflammatory process.

Acta Zoologica, Sep 11, 2014

Dudek, A., Sienkiewicz, W. and Kaleczyc, J. 2014. Contribution of the dorsal branch of the access... more Dudek, A., Sienkiewicz, W. and Kaleczyc, J. 2014. Contribution of the dorsal branch of the accessory nerve to the innervation of the trapezius muscle in the piga retrograde tracing study-Acta Zoologica (Stockholm) 00: 000-000. The study was aimed to establish the contribution of the dorsal branch of the accessory nerve (DBXI) to the innervation of the porcine trapezius muscle (TRAP). Combined retrograde tracing using fluorescent tracer Fast blue (FB) and surgical denervation procedure (excision of DBXI segment) were applied. FB+ neurons supplying the cervical (c-TRAP) and thoracic part (th-TRAP) of TRAP were localized in following nerve centres: the ipsilateral ventral horn of the grey matter of cervical neuromers, ipsilateral spinal ganglia and bilateral sympathetic chain ganglia. After the excision of DBXI segment, no FB+ motoneurons supplying c-TRAP were found while the mean number of those supplying th-TRAP was significantly decreased. A slight decrease in average numbers of sensory and autonomic neurons implemented in the innervation of both parts of TRAP was also observed. This study has revealed that in the pig DBXI is the only motor pathway to c-TRAP while the vast majority of motoneurons supplying porcine th-TRAP send their axons via DBXI.

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

PubMed, 1995

The presence and pattern of coexistence of catecholamine-synthesizing enzymes and some neuropepti... more 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.

Existence and coexistence of catecholamine-synthesizing enzymes and some neuropeptides in nerve fibres supplying the bovine ovary and its blood vessels

The preoptic area in the sexually immature pig. Morphological study

PubMed, 1993

The preoptic area was investigated in 5 immature pigs. This area is composed of three nuclei: lat... more The preoptic area was investigated in 5 immature pigs. This area is composed of three nuclei: lateral, medial, and median. The largest is the lateral nucleus.

Ontogeny of the expression of catecholamine synthesising enzymes in the female porcine median eminence arcuate nucleus complex (MEARC)

PubMed, 2003

The ontogeny of the catecholaminergic system of the median eminence (ME) arcuate nucleus (ARC) co... more The ontogeny of the catecholaminergic system of the median eminence (ME) arcuate nucleus (ARC) complex (MEARC) has been studied in various animal species but so far, nothing has been learnt about the development of catecholaminergic structures in the porcine MEARC. To study this problem the hypothalami from animals at different ages (six groups) were collected. Nerve structures immunoreactive (R) for the substances studied [(tyrosine hydroylase (TH), dopamine beta-hydroxylase (D(beta)H) and phenylethanoloamine-N-metylthransferase (PNMT)] were found in the pigs at different age periods. In MEARC, TH-IR structures appeared before the 70th day of foetal life, D(beta)H-IR before the 10th week of postnatal life and PNMT-IR only in sexually mature sows.

Distribution of neuropeptide Y-like immunoreactivity in the hypothalamic tuberal nuclei of the immature gilts

PubMed, 1994

The distribution of neuropeptide Y in the immature female pig hypothalamic tuberal nuclei was inv... more The distribution of neuropeptide Y in the immature female pig hypothalamic tuberal nuclei was investigated using avidin-biotin complex method. This study showed cell bodies containing NPY-immunoreactive material in the arcuate nucleus, dorsomedial and ventromedial nuclei, as well as internal layer of the median eminence and infundibulum, an observation not yet reported in the pig. NPY-immunoreactive fibers were noted throughout the hypothalamus, but most prominently within the periventricular region. Median eminence disclosed abundance of NPY-immunoreactive processes. The study shows a detailed organization of cell bodies and fibers containing NPY-immunoreactivity in tuberal hypothalamic nuclei of the immature gilts.

Immunohistochemical properties of sympathetic chain ganglia (SChG) neurons projecting to the porcine testis in animals subjected to hemicastration, castration and testosterone supplementation

PubMed, 2010

The present study was carried out on sexually mature boars. All the animals were injected with Fa... more The present study was carried out on sexually mature boars. All the animals were injected with Fast Blue into the right testis and then divided into four groups [(group 1--control (G1), group 2--hemicastreatad (G2), group 3 castrated (G3) and group 4--castrated and injected with testosterone (G4) boars)]. After a survival period of 3 weeks, G1 animals were transcardially perfused. In pigs of G2, right testis was removed, whereas in G3 and G4 animals both testes were removed. The pigs of G4 were injected with testosterone. After two weeks, all the animals were transcardially perfused and then their sympathetic chain ganglia were collected. The ganglia were cut into 12 microm-thick cryostat sections. The sections were stained using antisera against TH or DbetaH, VACHT or CHAT, NPY, VIP, GAL and AR. Testosterone plasma levels were evaluated with ELISA test. In control pigs, testosterone level amounted to 8.51 +/- 1.29 ng/ml, whereas in hemicastrated animals it was 1.72 +/- 0.35 ng/ml. Bilateral orchiectomy resulted in a drastic decline in testosterone level. Administration of exogenous hormone resulted in tremendous increase in its plasma level. In control pigs, FB-positive (FB+) neurons were found in the right and left ganglia. Immunohistochemical staining revealed that approximately 74% of FB+ neurons contained immunoreactivity to TH or DbetaH, whereas approximately 4% of FB+ cells were VACHT-positive. Among FB+/DbetaH+ neurons, 72% contained NPY and less than 1% stained for GAL. All FB+/VACHT+ neurons were also VIP+. Approximately 62% of FB+ somata expressed immunoreactivity to NPY, whereas 6% stained for VIP. In all experimental pigs, numbers of FB+ perikarya immunoreactive to TH (from 21% in G3, 26% in G2 to 29% in G4,) and DbetaH (62% in G2, 58% in G3 and 59% in G4,) were smaller than those found in G1 animals, whereas numbers of neurons displaying immunoreactivity to other substances studied were higher. The most significant increases in the number of immunoreactive cells were observed in regard to those expressing GAL (24% in G2, 30% in G3 and 27% in G4,) and VIP (11% in G2, 12% in G3 and 11% in G4,) whereas less distinct changes were observed in case of neurones which stained to NPY (67% in G2, 70% in G3 and 68% in G4,) and VAChT or ChAT (6% in G2, G3 and G4). The most apparent changes in the immunohistochemical features of FB+ neurons evoked by bilateral castration were observed in G3 pigs, whereas changes found in G4 animals were very similar to those observed in G2 pigs.

NADPH-diaphorase-containing cerebrovascular nerve fibres and their possible origin in the pig

PubMed, 1995

NADPH-diaphorase histochemical technique was applied to demonstrate the catalytic activity of nit... more 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.

Changes in the expression of some neuropeptides in the intestines and nerve ganglia during the porcine dysentery

Bulletin of The Veterinary Institute in Pulawy, 2005

Histochemistry and Cell Biology, 2020

The present study investigated the influence of castration performed at neonatal age on neuronal ... more The present study investigated the influence of castration performed at neonatal age on neuronal elements in the anterior pelvic ganglion of the male pig with immunohistochemistry and quantitative real-time PCR (qPCR). The ganglia were examined 3 and 6 months after surgery. In 3-month-old castrated pigs (3MCP) 74% of adrenergic and 31% of cholinergic neurons stained for caspase-3 (CASP-3), and much greater numbers of perikarya than in the control animals expressed CGRP, galanin (GAL) and VIP (peptides known to have neuroprotective properties). In 6-months-old castrated pigs (6MCP), an excessive loss (90%) of neurons and intraganglionic nerve fibres was found. The survived adrenergic and cholinergic neurons also expressed CASP-3, CGRP, GAL or VIP. The qPCR results corresponded with immunofluorescence findings. In 3MCP, genes for CASP-3 and CGRP were up-regulated, while the expression of those for DβH, VAChT, GAL, VIP and SP displayed statistically insignificant variations. In 6MCP, distinctly up-regulated were genes for CGRP, GAL, VIP, SP, DβH and VAChT, while the expression of casp3 gene was down-regulated. The study revealed for the first time the excessive loss of pelvic neurons following castration, and a realistic assumption is proposed, that the neurons died due to apoptosis triggered by androgen deprivation.