Papers by Waldemar Sienkiewicz
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
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.
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.
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.
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.
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.
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.
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.
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.
Acta histochemica, 1995
The distribution of neuropeptide Y, substance P, vasoactive intestinal polypeptide, Leu 5-enkepha... more The distribution of neuropeptide Y, substance P, vasoactive intestinal polypeptide, Leu 5-enkephalin, bombesin/gastrin-releasing peptide, calcitonin gene-related peptide, somatostatin, cholecystokinin and catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine-,8-hydroxylase was studied immunohistochemically in nerve fibres supplying the bovine vagina and uterus. The nerves containing tyrosine hydroxylase or dopamine-,8-hydroxylase and neuropeptide Y-immunoreactivity were particularly numerous in both organs. Substance P, vasoactive intestinal polypeptide and Leu 5-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.
Wspolczesna Onkologia-Contemporary Oncology, 2011
A Ai im m o of f t th he e s st tu ud dy y: : The purpose of this study was to investigate the pr... more A Ai im m o of f t th he e s st tu ud dy y: : The purpose of this study was to investigate the presence of vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and calcitonin gene related peptide (CGRP) neuropeptides in neoplastic tissue of the large intestine. The results obtained were compared to an unchanged part of the intestine originating from the same patient. M Ma at te er ri ia al l a an nd d m me et th ho od ds s: : The tissue samples were taken from the postoperative part of the intestine, resected during an operation due to treatment of colon cancer. Tissues were taken from 15 patients. Concentrations of neuropeptides were determined by ELISA tests, and results were calculated per 1 g of fresh tissue. The data obtained were statistically analysed. R Re es su ul lt ts s: : In tissues of the neoplastic tumour of the sigmoid colon and rectum the concentration of VIP and NPY was determined (28.9 ng/g and 8.6 ng/g, respectively). The observed values were statistically significantly lower than those observed in corresponding tissues of unchanged intestinal segments (83.9 ng/g for VIP and 19.5 ng/g for NPY). Surprisingly, amounts of CGRP were very similar in both neoplastic and intact tissues (13.3 ng/g and 13.4 ng/g respectively). C Co on nc cl lu us si io on ns s: : Immunoenzymatic investigations allow one to establish the concentration of neuropeptides associated with the enteric nervous system in cancer tissues of the large intestine. Various concentrations of these neuropeptides were observed in particular patients of both compared groups. The performed statistical analysis did not reveal associations between degree of cancer infiltration within the colonic wall and the level of presence of investigated neuropeptides in either cancer or control tissues. The fact of presence of these neuropeptides in cancer tissues confirmed by this study should be underlined at the same time. K Ke ey y w wo or rd ds s: : colon cancer, enteric nervous system, neuropeptides, calcitonin gene related peptide, neuropeptide Y, vasoactive intestinal polypeptide.
Acta Biologica Cracoviensia. Series Botanica. Supplement, 2008
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, Jun 1, 2015
Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide existing in two vari... more Pituitary adenylate cyclase activating polypeptide (PACAP) is a neuropeptide existing in two variant forms (of either 27 or 38 residues), widely present in numerous organs and evoking multiple effects both in the central and peripheral nervous systems. The present study was undertaken to evaluate the distribution pattern of PACAP-27 expression in the ovine pancreas. Using double immunohistochemical stainings co-localizations of PACAP-27 with galanin, SP or CRF were studied in intrapancreatic neurons. In intrapancreatic ganglia, immunoreactivty to PACAP-27 was found in 87.6 ± 5.4% of PGP 9.5-positive intrapancreatic neurons but not in intraganglionic nerve fibres. Numerous PACAP-27-immunoreactive nerve terminals were also observed between pancreatic acini and around small arterioles. No immunoreactivity to PACAP-27 was found in the endocrine pancreas. In 42.9 ± 6.2% of PACAP-27-immunoreactive intrapancreatic neurons the expression of galanin was also found. Statistically lower subpopulation (12.4 ± 4.0%) of intrapancreatic neurons exhibited simultaneously the immunoreactivity to PACAP-27 and SP. The expression of CRF was detected in the relatively smallest group (3.2 ± 1.4%) of PACAP-27-positive intrapancreatic neurons. The present results suggest that in the ovine pancreas PACAP-27 may play an important role as mediator of pancreatic functions. In PACAP-related pancreatic activities, a modulatory role of galanin, SP and to a lower extend of CRF is also likely.
Andrologia, Nov 25, 2010
This study was carried out on three adult male pigs of the large White Polish breed weighing 110-... more This study was carried out on three adult male pigs of the large White Polish breed weighing 110-130 kg each. The animals were anaesthetised and injected with retrograde tracer Fast Blue (FB) into right testis. Three weeks later, the pigs were deeply anaesthetised and perfused transcardially with fixative (4% paraformaldehyde in 0.1 m phosphate buffer pH 7.4). Collected ganglia were cut with freezing microtome into 12lm-thick sections. The sections were examined under a fluorescent microscope (Zeiss). FB-positive neurones were found in pelvic ganglia (anterior pelvic ganglion) (15.4% of all FB + neurones), prevertebral ganglia (caudal mesenteric, testicular, aortico-renal and renal ganglia) (59% of all FB + neurones), sympathetic chain ganglia (last four lumbar and first three sacral ganglia) (18.1% of all FB + neurones) and dorsal root ganglia (DRG) (first three lumbar and first three sacral ganglia) (7.4% of all FB + neurones). The majority of FB-positive nerve cell bodies were observed in ipsilateral ganglia, but they were also found in contralateral ganglia (approximately 85% and 15% respectively). Thus, FB-positive neurones were located in the left prevertebral, sympathetic chain and DRG, but surprisingly, they were absent in left anterior pelvic ganglia.
Cells, Aug 6, 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
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Papers by Waldemar Sienkiewicz