大鼠消化道神经激肽A免疫反应细胞的发生发育研究

大鼠消化道神经激肽Ａ免疫反应细胞的发生发育研究李力燕郭建辉蔡文琴＊（昆明医学院＊第三军医大学）微量元素与人体代谢本文应用免疫细胞化学ＰＡＰ法，系统地观察了胚胎１３－２１ｄ，生后１、３、５、１０、１５、２０、３０、６０、９０ｄ及成年大鼠食道、胃、小肠及...     

神经激肽A在大鼠胃发育中的定性定量分析

本研究用免疫组织化学PAP法和图像分析系统研究大鼠胚胎13d至成年胃中神经激肽A(NKA)的表达及变化。结果显示:胚胎14d,胃的肌间丛出现NKA免疫阳性反应,随发育相继在环肌、纵肌、粘膜下层、固有膜、粘膜肌及粘膜丛内出现NKA的表达,生后30d时NKA在胃的表达已具备成年鼠的分布特征。结果提示, (1)NKA在胃的表达变化主要在生前1周~生后4周内,其表达变化有两个关键时期,即生前1周~生后1周和生后1月末; (2)NKA对大鼠胃的发育可能具有重要的作用,可能与胃功能的建立密切相关。     

神经激肽A在大鼠结肠发育中的定性定量分析

探讨神经激肽A(NKA)在大鼠结肠发育中的表达及变化,应用免疫组织化学PAP法和图像分析系统观测大鼠胚胎13d至成年结肠中NKA的表达情况。结果显示:(1)在结肠,首先于胚胎17 d的肌间丛及上皮处出现NKA-IR的表达,随发育相继出现于纵肌、环肌、粘膜肌、固有膜、粘膜下层和粘膜丛,30 d时具备成年分布特征;(2)定量分析的结果与NKA~IR在结肠各层的变化一致;(3)NKA-IR细胞具有典型的肠道内分泌细胞的形态特征和分布特征。结果提示:(1)NKA在结肠的发生发育主要在生前5 d~生后4周内;(2)NKA在结肠的发生发育有两个关键时期,即生前5 d~生后1周和生后1月末;(3)NKA-IR细胞可能是肠道内分泌NKA的内分泌细胞;(4)NKA可能对大鼠结肠的发育具有重要作用,可能与结肠功能的建立密切相关。     

显示大鼠胃肌间神经丛的铺片方法

消化道全层辅片技术为研究消化道壁内神经丛的形态和分布提供了有效的实验手段［１］,由于胃形态不规则且胃壁肌层较厚,制作胃铺片标本十分困难,故以往的研究多集中于对肠道肌间神经丛的研究［２～４］,对胃肌间神经丛的形态研究主要以切片染色为主。为了研究工作的需要,作者对常规铺片技术进行适当改进,成功地制成胃纵肌层铺片标本。现以还原型铺酶ＩＩ黄逸酶（ＮＡＤＰＨ－ｄｉａｐｈｏｒａｓｅ,ＮＤＰ）组化染色显示大鼠胃肌间神经丛－氧化氮合酶（ｎｉｔｒｉｃｏｘｉｄｅｓｙｎｔｈａｓｅ,ＮＯＳ）阳性神经为例,将该方法介绍如…     

人胚胎结肠肠神经系统发育的观察

目的　研究人胚胎结肠肠神经系统的发育过程 ,为进一步研究先天性巨结肠的发病机制提供参考。　方法　采用一抗为蛋白基因产物 (protein gene product9.5 ,PGP9.5 )和 S- 10 0蛋白抗体的免疫组织化学 PAP法 ,显示结肠肠神经系统中的神经元和神经胶质。　结果　人胚胎结肠肠神经系统发育有明显的阶段性。在胚胎发育早期 (胎龄 2～ 3月 ) ,肠管壁发育差 ,以后出现菲薄的平滑肌层和低平的肠粘膜 ,此期偶在原始肌间神经丛位置见S- 10 0蛋白免疫反应性神经 ;至发育中期 (胎龄 4～ 5月 ) ,肠壁分化出 4层结构 ,出现相当发达的绒毛 ,肌间神经丛中细胞明显增多 ,呈弥散分布于整个肌层间并逐渐迁移到粘膜下层和粘膜层 ,由初级和次级突起构成复杂的神经网络 ;至晚期 (6～ 9月 ) ,肠壁各层均增厚 ,肌间神经丛成簇分布 ,神经纤维构成的网络出现更为细小的 3级突起 ,粘膜下神经丛分化形成浅丛和深丛。　结论　结肠神经系的发育具有明显的阶段性。发育早期神经开始在肠壁肌间丛位置出现 ,发育中期神经成分在肠壁各层中出现并发育增生 ,晚期肠壁各层神经已分化和成熟 ,而在不同的发育阶段 ,原始病因可导致临床表现不一的先天性巨结肠症     

神经激肽A在大鼠空肠发育中的定性定量分析

目的：探讨神经激肽A(NKA)在大鼠空肠发育中的表达及变化。方法：应用免疫组织化学PAP法和图像分析系统研究大鼠胚胎13d至成年空肠中NKA的表达情况。结果：(1)在空肠,首先于胚胎15d的肌间丛处出现NKA-IR的表达,随发育相继出现的纵肌、环肌、绒毛、小肠腺周、粘膜下层、深肌丛,生后30d已具备成年组的分布特征;(2)NKA-IR细胞具有典型的肠道内分泌细胞的形态特征和分布特征。结论：(1)NKA在空肠的发生发育主要在生前1周～生后4周内;(2)NKA在空肠的发生发育有2个关键时期,即生前1周～生后1周和生后1月末;(3)NKA-IR细胞可能是肠道内分泌NKA的内分泌细胞。     

扬子鳄胃的神经支配

采用组化方法对７只扬子鳄胃的神经支配进行了研究。结果如下：粘膜下层有胆碱酯酶（ＡＣｈＥ）阳性反应神经节和神经纤维相互交织组成的神经丛，一些神经纤维穿过粘膜在基底膜附近形成次组丛，终止在腺细胞。在环行肌和纵行肌之间有丰富的ＡＣｈＥ阳性反应神经节和神经丛。在肌层内胆碱能神经的走向与平滑肌细胞的长轴平行。肾上腺素能神经只见于肌层，神经纤维同样与平滑肌细胞长轴平行。     

大鼠空肠肌间神经丛一氧化氮合酶的组化光镜及电镜观察

大鼠空肠肌间神经丛一氧化氮合酶的组化光镜及电镜观察肖岚，蔡文琴，孙榆第三军医大学重庆６３００３８正常Ｗｉｓｔａｒ成年大鼠５只，行空肠纵肌层铺片，用ＮＡＤＰＨ－黄递酶（ＮＤＰ）组化染色以对肌间神经丛－氧化氮合酶（ＮＯＳ）的分布进行光镜及电镜观察。结果显...     

小鼠脾发育分化的组织学及组织化学观察

观察了ＢＡＬＢ／ｃ小鼠胚胎至成年脾发育分化的组织学和组织化学变化。（１）胚胎１２天，胃背系膜内出现由间充质细胞形成的脾原基。此后，脾窦、网状细胞及淋巴细胞相继出现；生后１天出现动脉周围淋巴鞘；以后逐渐形成脾小结。（２）胚胎１３天时脾中出现糖原和ＲＮＡ颗粒，并渐增多，生后１周糖原量骤减，２周至成年为阴性。ＲＮＡ则在白髓中少，红髓中多。（３）碱性磷酸酶在１７天胚胎脾内出现阳性细胞，以后增多。（４）酸性     

NOS在大鼠胃肠肌间神经丛的分布模式

ＮＯＳ在大鼠胃肠肌间神经丛的分布模式肖岚蔡文琴（第三军医大学组胚教研室）本研究应用ＮＡＤＰＨ－ｄ组化染色法和图像分析技术对成年大鼠胃肠肌间神经丛中ＮＯＳ的分布进行观察，以期得到大鼠胃肠肌间神经丛的构筑模式，结果如下：①胃：各部分神经丛网格形态、密度、...     

An immunohistochemical study of the distribution of enteric GABA-containing neurons in the rat and guinea-pig intestine

gamma-Aminobutyric acid (GABA) antiserum was applied to sections of rat and guinea-pig intestine which were subsequently processed to reveal any immunoreactivity using either fluorescence or peroxidase techniques. Immunopositive fibres were demonstrated in stomach, duodenum, ileum and colon of rat and guinea-pig intestine. Myenteric ganglia and nerve bundles in the circular muscle contained immunopositive nerve fibres, while the longitudinal muscle, submucosa and mucosa were only rarely innervated. In favourable sections, immunopositive fibres could be seen running from the myenteric plexus into the circular muscle, thus suggesting that the GABA-immunopositive nerves in the circular muscle originate from neurons in the myenteric plexus. In both rat and guinea-pig, immunoreactive nerve cell bodies were most numerous in the myenteric plexus of the colon. In the rat, immunopositive fibres in the circular muscle were most abundant in the ileum, whereas in the guinea-pig it was the colon circular muscle that was most richly innervated. The results demonstrate that neurons which show GABA immunoreactivity are present along the length of the gastrointestinal tract. Their distribution in both myenteric ganglia and circular muscle is heterogeneous both within and between the two species studied. It is probable that this heterogeneity reflects the diversity and specificity of function of this class of enteric neurons.     

神经激肽A在大鼠回肠发育中的定位定量分析

目的探讨神经激肽A(NKA)在大鼠回肠发育中的表达及变化。方法应用免疫组织化学PAP法和图像分析系统研究大鼠胚胎13d至成年回肠中NKA的表达。结果1．在回肠,首先于胚胎17d的肌间神经丛处出现NKA-IR的表达,随发育相继出现在纵肌层、环肌层、绒毛、小肠腺周、黏膜下层、深肌丛,生后30d已具备成年组的分布特征;2．定量分析中数据的变化与NKA-IR在回肠各层阳性强弱的变化一致;3．NKA-IR细胞具有典型的肠道内分泌细胞的形态特征和分布特征。结论1．NKA在回肠的发生发育主要在生前1周～生后4周内(但绒毛内神经纤维的密度于60d时才达成年水平);2．NKA在回肠的发生发育有两个关键时期,即生前1周～生后1周和生后1月末;3．NKA-IR细胞可能是肠道内分泌NKA的内分泌细胞;4．NKA对大鼠回肠的发育具有重要的作用,可能与回肠功能的建立密切相关。     

Neurofilament and glial fibrillary acid protein-related immunoreactivity in rodent enteric nervous system

Using antisera raised against neurofilaments and the glial fibrillary acidic protein (GFAP) we have examined the appearance and distribution of neurofilament- and GFAP-like immunoreactivity in the enteric nervous system of rat, mouse and guinea-pig. In whole mounts of the external circular and longitudinal muscle layers, including the myenteric plexus, a high number of neurofilament-positive perikarya were visualized both in the ganglia and in the circularly running interconnecting strands in all three species. These cells were large, usually with eccentrically placed nuclei and single, relatively thick neurofilament-positive processes. In addition, in guinea-pig myenteric plexus a small number of cells with multiple processes could be seen. Both in the longitudinal and circular interconnecting strands a large number of thin, smooth, neurofilament-positive fibres were observed. This regular network of ganglia and strands was superimposed on a sparse system of thin, usually individual neurofilament-positive fibres in the underlying circular muscle layer. Cryostat sections revealed neurofilament-positive cell bodies in the submucous plexus, whereas fibres showing neurofilament-like immunoreactivity were observed in all layers of the gut wall, with the exception of the epithelium. In whole mounts including rat and mouse myenteric plexus, a large number of cells and fibres showing GFAP-like immunoreactivity were visualized. The GFAP-positive cells were smaller and more numerous than the neurofilament-positive ones. They were present both within the ganglia and in the interconnecting strands. Several short fluorescent processes could frequently be seen emanating from the cell body. Both the strands and the ganglia contained a high number of thin, GFAP-positive fibres. Fluorescent fibres and cells were also observed in the circular muscle layer. In sections of rat and mouse small intestine, cells were observed throughout the gut wall, with the exception of the epithelium. Double labelling experiments clearly showed that neurofilament- and GFAP-positive cells represented separate cell populations. Furthermore, GFAP-positive cells and fibres outlined the neurofilament-positive perikarya. It is thus likely that the GFAP-positive cells represent enteric glial cells. The pre- and postnatal development of neurofilament- and GFAP-like immunoreactivity was studied in whole mounts from rat embryos and pups. Furthermore, the presence of neurofilament and GFAP-positive fibres was observed in whole mount preparations of rat and mouse mesenterium.(ABSTRACT TRUNCATED AT 400 WORDS)     

Neuronal substance P in the esophagus. Distribution and effects on motor activity

Substance P-immunoreactive nerve fibres were fairly numerous in the lower esophagus of the guinea-pig and cat but few in the pig. They were particularly numerous in the myenteric and submucosal plexuses but could be detected also in the circular and longitudinal smooth muscle and in the muscularis mucosae. Only in the cat were SP-immunoreactive cell bodies detected, albeit in low number, in the myenteric plexus. Radioimmunoassay showed that the lower part of the cat esophagus contained approximately 10 times more immunoreactive SP than the upper part and that the muscle layer contained more SP than the mucosa. Motor effects of synthetic SP were studied on segments from circular smooth muscle of cat esophagus. SP contracted the smooth muscle and enhanced the response to electrical stimulation. These effects of SP could be blocked by the specific SP antagonist (d -Pro,²d -Trp^7,8)-SP. The contractile response to electrical stimulation could be blocked by the cholinergic muscarinic blocker atropine and the opiate receptor agonist leu-enkephalin but not by the SP antagonist or by adrenergic blockers. Hence, the results suggest that cholinergic neurons innervate the circular smooth muscle, and that opiate receptor agonists suppress transmission in these neurons. Neuronal SP in the esophagus may serve to enhance the contractile responses of esophageal smooth muscle.     

Nitric oxide synthase immunoactivity and NADPH diaphorase enzyme activity in neurons of the gastrointestinal tract of the toad, Bufo marinus.

The presence of nitric oxide synthase (NOS) was demonstrated immunohistochemically, and NADPH diaphorase was demonstrated by enzyme histochemistry in neurons throughout the gastrointestinal tract of the anuran amphibian, Bufo marinus. Successive staining showed that NOS immunoreactivity and NADPH diaphorase activity occurred in precisely the same subgroup of enteric neurons. Subsequent detailed studies of the distribution of these neurons were made using NADPH diaphorase histochemistry. Numerous reactive nerve cell bodies and fibres were found in the myenteric plexus from the esophagus to the cloaca. A dense innervation of the longitudinal and circular muscle layers occurred throughout the gastrointestinal tract. The lamina muscularis mucosae was only prominent in the stomach, where it was sparsely innervated. Reactive nerve cell bodies were common in the submucosa of the large intestine, less common in the small intestine and extremely rare in the stomach and esophagus. Reactive fibres contributed to subepithelial plexuses in the esophagus, colon, rectum and cloaca. It is concluded that NOS/NADPH diaphorase is conserved amongst vertebrate classes and that NO is a likely neurotransmitter in the toad gastrointestinal tract.     

Glomerular handling of immune complex in the acute phase of active in situ immune complex glomerulonephritis employing cationized ferritin in rats

The distribution and co-localization of nitric oxide synthase (NOS) and vasoactive intestinal polypeptide (VIP) were examined by means of immunohistochemistry and NADPH diaphorase (NADPH-d) histochemistry in the gut of patients with Hirschsprung's disease. In the normoganglionic segment, many nitrergic nerve cells were localized in Auerbach's plexus and nerve fibres were observed preferentially in the circular muscle. The submucosal nitrergic nerve cells were mainly situated in Schabadasch's plexus with occasional cells demonstrable in Meissner's plexus. NOS and VIP were co-localized in most ganglion cells of Auerbach's plexus. In the oligoganglionic segment, a marked reduction of NOS- and VIP- positive nerve cells and fibres was noticed in both the myenteric and submucosal plexuses, and nitrergic fibres had disappeared in the inner layer of the circular muscle. In the aganglionic segment, NOS and VIP were revealed only in extrinsic nerve fasciculi and rami and co-localized in a few fibres. From these observations, the inner layer of the circular muscle of the oligoganglionic segment and the whole of the muscularis propria of the aganglionic segment were considered to be totally lacking in nitrergic innervation. Nitrergic nerves of the human colon comprise both intrinsic and extrinsic elements and the majority of intrinsic nitrergic nerve cells contain VIP. Very low numbers of extrinsic nitrergic fibres contain VIP.     

A light microscope study of the distribution of muscle in the frog esophagus and stomach.

The present study reports light microscopical observations of the distribution of muscle in the esophagus and stomach of both the bull frog (Rana catesbeiana) and the African clawed frog (Xenopus laevis). The external muscle coat of the upper half of the esophagus in both species had several collagen coated bundles of striated muscle fibres around the circumference. These striated muscle bundles ran longitudinally from the pharynx to around the vicinity of the center of the esophagus. Beneath these striated muscle bundles was an inner circular layer of smooth muscle. In both species, the inner circular layer of smooth muscle was particularly thick in the region close to the pharynx. In the bull frog, the lower half of the esophagus lacked striated muscle. However, the circular smooth muscle layer, extending from the upper half of the esophagus, was also observed throughout the lower half of the esophagus. An outer longitudinal layer of smooth muscle developed towards the terminal portion of the esophagus such that in this region, both outer longitudinal and inner circular layers of smooth muscle were observed. Similarly in the African clawed frog, the inner circular layer of smooth muscle was continuous along the full length of the esophagus. Again, no striated muscle bundles were observed in the lower half of the esophagus. However, the outer longitudinal layer of smooth muscle was seen to develop in the middle region of the esophagus. Its muscle layer extended to the terminal portion of the esophagus. Thus, both outer longitudinal and inner circular layers of smooth muscle were observed throughout the lower half of the esophagus. In both frogs, the thickness of the outer longitudinal and inner circular layers of smooth muscle changed before and after the esophago-gastric junction. In both frogs, no muscularis mucosa was observed in the esophageal wall. However, in the lower half of the esophagus of the African clawed frog, small bundles of smooth muscle were observed here and there in the submucosa. A fully developed muscularis mucosa with both outer longitudinal and inner circular layers was observed in the upper stomach of both frogs.