scholarly journals Proliferative enteropathy (PPE)-induced changes in the expression of DBH, VAChT and NOS in the neurons of intramural ganglia of the porcine ileum

2008 ◽  
Vol 53 (No. 10) ◽  
pp. 533-542 ◽  
Author(s):  
Z. Pidsudko ◽  
K. Wasowicz ◽  
J. Kaleczyc ◽  
M. Majewski ◽  
M. Lakomy
Keyword(s):  
Enteric Neurons ◽  
Double Labelling ◽  
Lawsonia Intracellularis ◽  
Large White ◽  
Submucous Plexus ◽  
Chemical Coding ◽  
Proliferative Enteropathy ◽  
Induced Changes ◽  
Oxide Synthase ◽  
Intramural Ganglia

As enteric neurons are regarded to be highly adaptive in their response to various pathological states, including inflammation, it appears to be of interest to study the chemical coding of neurons in the intramural ganglia of the ileum wall in the course of porcine proliferative enteropathy (PPE) evoked by <I>Lawsonia intracellularis.</I> The study was performed on 12 juvenile pigs of the Large White Polish breed. The pigs were divided into the control (C, <I>n</I> = 6) group and the group consisting of pigs with clinically diagnosed <I>Lawsonia intracellularis</I> infection (E, <I>n</I> = 6). In E group animals the infection was confirmed with a PCR-based test. All the animals were sacrificed and segments of the ileum being pathologically changed were processed for double-labelling immunofluorescence using antibody against protein gene-product 9.5 (PGP 9.5) combined with antibody for dopamine &beta;-hydroxylase (D&beta;H), vesicular acetylcholine transporter (VAChT) or nitric oxide synthase (NOS). Immunohistochemistry revealed in the inner submucous plexus (ISP) and outer submucous plexus (OSP) an increase of the number of neurons containing D&beta;H and VAChT in the E group. Interestingly, a decrease in the number of D&beta;H- and VAChT-positive neurons in meynteric plexus (MP) ganglia of the E group animals was observed. The most remarkable difference in the chemical coding of enteric neurons between the control and PPE-suffering pigs was a significant increase of the number of NOS-positive nerve cells in the MP and OSP of the infected animals. The present results show that acetylcholine, catecholamines and NO may be involved in the regulation of functions of the porcine enteric nerve pathways not only under physiological, but also pathological conditions.

Characterisation of cocaine- and amphetamine- regulated transcript-like immunoreactive (CART-LI) enteric neurons in the porcine small intestine

2012 ◽  
Vol 60 (3) ◽  
pp. 371-381 ◽  
Author(s):  
Joanna Wojtkiewicz ◽  
Sławomir Gonkowski ◽  
Marek Bladowski ◽  
Mariusz Majewski
Keyword(s):  
Nitric Oxide ◽  
Small Intestine ◽  
The Other ◽  
Enteric Neurons ◽  
Double Labelling ◽  
Immunofluorescence Technique ◽  
Submucous Plexus ◽  
Oxide Synthase ◽  
First Time ◽  
Labelling Immunofluorescence

The aim of this study was to investigate the distribution and the number of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons and the co-localisation of CART with substance P (SP), somatostatin (SOM), nitric oxide synthase (NOS) and vasoactive intestinal polypeptide (VIP) within the enteric nervous system (ENS) in the porcine small intestine. Accordingly, the myenteric plexus (MP), outer submucous plexus (OSP) and inner submucous plexus (ISP) of the small intestine (duodenum, jejunum and ileum) were studied by double-labelling immunofluorescence technique. CART-LI neurons were observed in all gut fragments and all types of intramural plexuses studied and amounted from 0.2 ± 0.1% in the ISP of ileum to 22.4 ± 2.4% in the MP of this segment. The co-localisation of CART and NOS or/and VIP was observed depending on the segment of the gut and the complexity of the intramural plexus. On the other hand, during this study the co-localisation of CART and SOM or/and SP was not observed. The present study, for the first time, presents a detailed description of the CART distribution pattern and co-localisation with other neuromodulators within the ENS of the porcine small intestine.

Immunohistochemical characteristics and distribution of neurons in the intramural ganglia supplying the urinary bladder in the male pig

2013 ◽  
Vol 16 (4) ◽  
pp. 629-638 ◽  
Author(s):  
Z. Pidsudko
Keyword(s):  
Urinary Bladder ◽  
Large Population ◽  
Double Labelling ◽  
Chemical Coding ◽  
Visceral Peritoneum ◽  
Calcitonin Gene ◽  
Oxide Synthase ◽  
Cryostat Sections ◽  
Labelling Immunofluorescence ◽  
Intramural Ganglia

Abstract This study investigated the distribution and chemical coding of neurons in intramural ganglia of the urinary bladder trigone (UBT-IG) and cervix (UBC-IG) in the male pig using combined retrograde tracing and double-labelling immunohistochemistry. Additionally, immunoblotting was used to confirm the presence of marker enzymes for main populations of autonomic neurons. Retrograde fluorescent tracer Fast Blue (FB) was injected into the wall of both the left and right side of the bladder trigone, cervix and apex during laparotomy performed under thiopental anaesthesia. Twelve μm-thick cryostat sections were processed for double-labelling immunofluorescence with antibodies against tyrosine hydroxylase (TH), dopamine β-hydroxylase (DBH), neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), calcitonin gene-related peptide (CGRP), substance P (SP) and vesicular acetylcholine transporter (VAChT). UBT-IG and UBC-IG neurons in both parts of the organ formed characteristic clusters (from few to tens of neuronal cells) found under visceral peritoneum or in the outer muscular layer. Immunohistochemistry revealed several subpopulations in UBT-IG and UBC-IG neurons, namely noradrenergic (ca. 76% and 76%), cholinergic (ca. 22% and 20%), non-adrenergic/non-cholinergic nerve cells (ca. 1.5% and 3.8%), NPY- (ca. 66% and 58%), SOM- (ca. 39% and 39 %), VIP- (ca. 5% and 0%) and NOS- immunoreactive (IR) (ca. 1.5% and 3.8%), respectively. Immunoblotting using antibodies to TH and VAChT showed the presence of studied proteins as revealed by the presence of protein bands of the correct molecular weight. This study has revealed a relatively large population of differently coded UBT- and UBC- IG neurons, which constitute an important element of the complex neuroendocrine system involved in the regulation of the male urogenital organs function.

scholarly journals Immunohistochemical characterisation of neurons in the mandibular ganglion and nerve fibres supplying the porcine mandibular gland

2017 ◽  
Vol 61 (No. 7) ◽  
pp. 361-373 ◽  
Author(s):  
M. Klimczuk ◽  
P. Podlasz ◽  
W. Sienkiewicz ◽  
A. Franke-Radowiecka ◽  
A. Dudek ◽  
...  
Keyword(s):  
Mandibular Gland ◽  
Double Labelling ◽  
Nerve Terminals ◽  
Rt Pcr ◽  
Nerve Fibres ◽  
Chemical Coding ◽  
Pcr Product ◽  
Calcitonin Gene ◽  
Oxide Synthase ◽  
Cryostat Sections

The present study was designed to investigate the chemical coding of neurons in the mandibular ganglion (MGn) and nerve fibres supplying the porcine mandibular gland (MGl) with the use of immunofluorescence and RT-PCR. The cryostat sections from MGn and MGl were processed for double-labelling immunohistochemistry using antisera against vesicular acetylcholine transporter (VAChT), choline acetyltransferase (ChAT), dopamine β-hydroxylase (DβH), neuronal nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), galanin (GAL), substance P (SP) and calcitonin gene-related peptide (CGRP). The MGl was found to be richly supplied by VAChT-positive nerve fibres that surrounded intra- and interlobular salivary ducts. A large number of VAChT-immunoreactive (VAChT-IR) nerve terminals were also observed around acini. Many periductal and periacinar nerve fibres stained positive for DβH. Immunoreactivity to GAL, NPY or VIP was observed in an intermediate number of nerve terminals which were associated with both salivary ducts and acini. Double-immunostaining revealed that in MGn nearly all neurons stained positive for VAChT/ChAT (98.45 ± 0.59%, mean ± SEM) and nNOS (99.71 ± 0.18%). An intermediate number of the nerve cell bodies displayed immunoreactivity to NPY or VIP (18.67 ± 0.52% and 8.11 ± 0.36%, respectively). Single GAL-IR and CGRP-positive neurons were also observed. RT-PCR revealed the presence of transcripts of ChAT, VAChT, nNOS, NPY, VIP and GAL. For SP and DβH very weak signals were observed. RT-PCR with primers targeting CGRP did not generate any PCR product.

scholarly journals Alterations in Galanin-Like Immunoreactivity in the Enteric Nervous System of the Porcine Stomach Following Acrylamide Supplementation

2019 ◽  
Vol 20 (13) ◽  
pp. 3345 ◽  
Author(s):  
Katarzyna Palus ◽  
Krystyna Makowska ◽  
Jarosław Całka
Keyword(s):  
Nervous System ◽  
Enteric Nervous System ◽  
Daily Intake ◽  
Enteric Neurons ◽  
Double Labelling ◽  
Recovery Processes ◽  
High Doses ◽  
Oxide Synthase ◽  
The Impact ◽  
Double Labelling Immunohistochemistry

In recent years, a significant increase in the consumption of products containing large amounts of acrylamide (e.g., chips, fries, coffee), especially among young people has been noted. The present study was created to establish the impact of acrylamide supplementation, in tolerable daily intake (TDI) dose and a dose ten times higher than TDI, on the population of galanin-like immunoreactive (GAL-LI) stomach neurons in pigs. Additionally, in the present study, the possible functional co-operation of GAL with other neuroactive substances and their role in acrylamide intoxication was investigated. Using double-labelling immunohistochemistry, alterations in the expression of GAL were examined in the porcine stomach enteric neurons after low and high doses of acrylamide supplementation. Generally, upregulation in GAL-LI immunoreactivity in both myenteric and submucous plexuses was noted in all stomach fragments studied. Additionally, the proportion of GAL-expressing cell bodies simultaneously immunoreactive to vasoactive intestinal peptide (VIP), neuronal nitric oxide synthase (nNOS) and cocaine- and amphetamine- regulated transcript peptide (CART) also increased. The results suggest neurotrophic or/and neuroprotective properties of GAL and possible co-operation of GAL with VIP, nNOS, CART in the recovery processes in the stomach enteric nervous system (ENS) neurons following acrylamide intoxication.

scholarly journals Aspirin Administration Affects Neurochemical Characterization of Substance P-Like Immunoreactive (SP-LI) Nodose Ganglia Neurons Supplying the Porcine Stomach

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Liliana Rytel ◽  
Jarosław Całka
Keyword(s):  
Substance P ◽  
Control Group ◽  
Double Labelling ◽  
Large White ◽  
Nodose Ganglia ◽  
Calcitonin Gene ◽  
Mucosal Layer ◽  
Inflammatory Processes ◽  
Oxide Synthase

Background. Acetylsalicylic acid (ASA) is a commonly used anti-inflammatory, antipyretic, and analgesic drug, which has many side effects on the gastric mucosal layer. Despite this, knowledge concerning the influence of ASA on neuronal cells supplying the stomach is very scanty. Methods. This investigation was performed on ten immature gilts of the Large White Polish race divided into two groups (five animals in each): a control group and animals which were treated with ASA. The retrograde neuronal tracer Fast Blue (FB) was injected into the prepyloric region of the stomach in all animals. ASA was then given orally to the experimental (ASA) group of gilts from the seventh day after FB injection to the 27th day of the experiment. After this period, all animals were euthanized. Immediately after euthanasia, nodose ganglia (NG) were collected and subjected to a standard double-labelling immunofluorescence technique using antibodies directed toward substance P (SP) and other selected neuronal factors, such as galanin (GAL), neuronal isoform of nitric oxide synthase (nNOS), vasoactive intestinal polypeptide (VIP), and calcitonin gene-related peptide (CGRP). Key Results. The obtained results show that SP-LI neurons located in NG supplying the porcine stomach were also immunoreactive to all the above-mentioned neuronal factors. Moreover, ASA administration caused an increase in the degree of colocalization of SP with other neuronal active substances, and the most visible changes concerned the number of neurons simultaneously immunoreactive to SP and CGRP. Conclusions and Inferences. These observations indicate that the population of SP-LI neurons supplying the stomach is not homogeneous and may undergo changes after ASA administration. These changes are probably connected with inflammatory processes and/or neuroprotective reactions although their exact mechanisms remain unknown.

Distribution and chemical coding patterns of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons in the enteric nervous system of the porcine stomach cardia

2015 ◽  
Vol 18 (3) ◽  
pp. 515-522 ◽  
Author(s):  
W. Rękawek ◽  
P. Sobiech ◽  
S. Gonkowski ◽  
K. Żarczyńska ◽  
A. Snarska ◽  
...  
Keyword(s):  
Nervous System ◽  
Enteric Nervous System ◽  
Neuronal Nitric Oxide Synthase ◽  
Nerve Fibers ◽  
Vesicular Acetylcholine Transporter ◽  
Double Labelling ◽  
Dense Network ◽  
Chemical Coding ◽  
Oxide Synthase ◽  
Labelling Immunofluorescence

Abstract The aim of this study was to determine the presence of cocaine- and amphetamine-regulated transcript-like immunoreactive (CART-LI) neurons and co-localisation of CART with vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (n-NOS), vasoactive intestinal polypeptide (VIP), substance P (SP) and leu-enkephalin (LENK) in the enteric nervous system of the porcine gastric cardia by using a double-labelling immunofluorescence technique. CART-LI neurons were observed in the myenteric plexus (18.2±2.6%). A dense network of CART-LI nerve fibers was mainly observed in the muscular layer. CART showed co-localization mainly with VAChT, n-NOS, VIP and to a lesser degree with LENK and SP. Distribution of CART and its co-localization with other neurotransmitters suggest that this peptide plays an important role in gastric motility in the pig.

Distribution pattern and chemical coding of neurons of the sympathetic chain ganglia supplying the descending colon in the pig

2010 ◽  
Vol 58 (2) ◽  
pp. 189-198 ◽  
Author(s):  
Cezary Skobowiat ◽  
Jarosław Calka ◽  
Krzysztof Wasowicz ◽  
Mariusz Majewski
Keyword(s):  
Great Majority ◽  
Retrograde Tracing ◽  
Sympathetic Chain ◽  
Double Labelling ◽  
Chemical Coding ◽  
Pituitary Adenylate Cyclase ◽  
Descending Colon ◽  
Oxide Synthase ◽  
Ventral Area ◽  
Labelling Immunofluorescence

Sympathetic chain ganglia (SChG) neurons projecting to the descending colon of the pig were studied by means of retrograde tracing (Fast Blue, FB) and double-labelling immunofluorescence methods. FB was injected into the gut wall and after three weeks survival time the animals were transcardially perfused with paraformaldehyde and the bilateral sympathetic trunks were collected. The FBpositive neurons were localised only in the lumbar (L 1 –L 5 ) ganglia of the sympathetic trunk and appeared either as small (30–50 μm in diameter) round-shaped perikarya forming clusters localised in caudal-ventral area or, rarely, as bigger (50–80 μm) and dispersed solitary irregular perikarya. Immunohistochemical staining revealed the catecholaminergic (tyrosine hydroxylase-/dopamine β-hydroxylase-immunoreactive) character of the great majority of FB-positive neurons which preferentially co-expressed neuropeptide Y. In addition, none of the FB-positive perikarya was immunopositive to galanin, somatostatin, choline acetyltransferase, vasoactive intestinal peptide, pituitary adenylate cyclase-activating peptide, leu 5 -enkephalin, nitric oxide synthase, substance P and calcitonin-generelated peptide.

The effect of I.V. L-NG Methylarginine Hydrochloride (L-NMMA: 546C88) on Basal and Acetazolamide (Diamox®) induced Changes of Blood Velocity in Cerebral Arteries and Regional Cerebral Blood flow in Man

Cephalalgia ◽  
2005 ◽  
Vol 25 (5) ◽  
pp. 344-352 ◽  
Author(s):  
LH Lassen ◽  
B Sperling ◽  
AR Andersen ◽  
J Olesen
Keyword(s):  
Blood Flow ◽  
Cerebral Blood Flow ◽  
Regional Cerebral Blood Flow ◽  
Cerebral Arteries ◽  
Blood Velocity ◽  
Percentage Increase ◽  
Regional Cerebral Blood ◽  
Cerebral Arterioles ◽  
Induced Changes ◽  
Oxide Synthase

The aim of this study was to estimate the effect of Nitric Oxide synthase (NOS)-inhibition (L-NMMA) on the diameter of the middle cerebral artery (MCA) and on regional cerebral blood flow (rCBF). Furthermore, to assess the effect of L-NMMA on acetazolamide induced increases in MCA blood velocity (Vmean) and rCBF. In an open crossover design 12 healthy subjects attended the laboratory twice. The first day 6 mg/kg L-LNMMA i.v. over 15 min preceded 1 g acetazolamide iv over 5 min. Eight days later only acetazolamide was given. Vmean in MCA was determined with transcranial Doppler (TCD) and rCBF with Xe-133 inhalation SPECT at baseline, after L-NMMA and 25 and 55 min after acetazolamide infusion. After L-NMMA the decrease in rCBFMCA was 6.8% (± 7.4) ( P < 0.019, n = 12), whereas Vmean was not affected ( P = 0.83, n = 8). The change in MCA diameter was estimated to -1.3% ( P = 0.44, n = 8). L-NMMA did not affect acetazolamide increases in Vmean ( P = 0.67, n = 8) nor rCBF ( P = 0.29, n = 12). The percentage increase of Vmean was 1.5 times that of rCBF ( n = 8). Our data suggest that the basal tone of human cerebral arterioles but not of conduit arteries is NO-dependent. The action of acetazolamide in man is not NO-dependent.

scholarly journals Draft Genome Sequence of Lawsonia intracellularis Strain E40504, Isolated from a Horse Diagnosed with Equine Proliferative Enteropathy

2017 ◽  
Vol 5 (19) ◽  
Author(s):  
Nandita S. Mirajkar ◽  
Molly R. Kelley ◽  
Connie J. Gebhart
Keyword(s):  
Genome Sequence ◽  
Genomic Island ◽  
Draft Genome ◽  
Draft Genome Sequence ◽  
Lawsonia Intracellularis ◽  
Protein Coding ◽  
Content Type ◽  
Obligate Intracellular ◽  
Proliferative Enteropathy ◽  
Rna Genes

ABSTRACT Reported herein is the draft genome sequence of equine-origin Lawsonia intracellularis strain E40504, an obligate intracellular bacterium and the etiological agent of equine proliferative enteropathy. The 1.69-Mb draft genome sequence includes 1,380 protein-coding genes and 49 RNA genes, and it lacks a genomic island reported in swine-origin L. intracellularis strain PHE/MN1-00.

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