Kaleczyc J, Klimczuk M, Franke-Radowiecka A, Sienkiewicz W, Majewski M, Łakomy M
Department of Functional Morphology, Faculty of Veterinary Medicine, University of Warmia and Mazury, Olsztyn, Poland.
Anat Histol Embryol. 2007 Jun;36(3):186-93. doi: 10.1111/j.1439-0264.2006.00744.x.
The 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), Leu(5)-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.
本研究旨在调查正常（对照）猪和患痢疾猪的胃壁内神经元生物活性物质的表达情况。使用了八只幼年雌性猪。对患痢疾的动物（n = 4；接种猪痢疾短螺旋体）和对照动物（n = 4）均进行深度麻醉，经心内灌注缓冲多聚甲醛，并收集包括胃底壁各层的组织样本。对冰冻切片进行双重标记免疫荧光处理，以研究壁内神经结构的分布（用抗蛋白基因产物9.5抗体显示）及其化学编码，所用抗体包括抗囊泡乙酰胆碱（ACh）转运体（VAChT）、一氧化氮合酶（NOS）、甘丙肽（GAL）、血管活性肠肽（VIP）、生长抑素（SOM）、亮氨酸脑啡肽（LENK）、P物质（SP）和降钙素基因相关肽（CGRP）。在对照猪的内、外黏膜下神经丛中，大多数神经元为SP阳性（分别为55%和58%）或VAChT阳性（54%）。许多神经元也被CGRP（43%和45%）或GAL（20%和18%）染色，单个核周体为NOS、SOM或VIP阳性。肌间神经丛神经元被NOS（20%）、VAChT（15%）、GAL（10%）、VIP（7%）、SP（6%）或CGRP（单个神经元）染色，但它们为SOM阴性。未发现对LENK有免疫反应的壁内神经元。对照猪和患痢疾猪的肠神经元化学编码最显著的差异是，感染动物黏膜下神经丛中GAL和VAChT阳性神经细胞数量大幅增加（分别高达61%和85%）。目前的结果表明，在猪痢疾病程中，GAL和ACh在发炎的猪胃局部神经回路中具有特定作用。
