人胎食管NOS阳性神经元发育的研究

目的：探讨人胎食管壁内NOS阳性神经元的发育。方法：用NADPH－d组织化学法对人胎食管NOS阳性神经元的分化,迁移和发育进行了观察。结果：第4－5月龄时,肌间神经节处的圆形细胞出现一氧化氮合酶阳性反应,并分化演变成梭形的NOS阳性神经细胞,呈条索状排列沿内环肌与外纵肌层之间迁移,有的穿过内环肌层进入粘膜下层,第6个月龄时,迁移到靶细胞附近的NOS阳性神经元胞体明显增大,突起伸长,在肌层和粘膜下层出现NOS阳性神经纤维分布。结论：人胎食管NOS阳性神经元来源于胚胎早期神经节处的圆形细胞,通过分化,迁移,生长发育形成成熟的NOS阳性神经元。     

先天性巨结肠病肠壁NOS阳性神经元光镜和电镜观察

目的：探讨NOS阳性神经元与先天性巨结肠病病因及病理机制的关系。方法：对扩张段和移行段肠壁分别作全层铺片,NADPH－d酶组织化学染色,光镜和扫描电镜下观察NOS阳性神经结构。结果：扩张肠段光镜下肠肌丛神经节和神经元均较大,节内神经元染色深数量多,沿神经节周边及神经纤维发出处排列。扫描电镜下神经元胞体较大,排列较密,发出的神经纤维较多,在各个方向上相互连接。沿肌纤维排列的神经元之间有较多的横向连接纤维,肠肌丝社会元还通过穿行于环行肌层的神经纤维和粘膜下层神经元相连接,移行段光镜下节内神经元胞浆染色较淡,深浅不一,神经节和神经元均较小,发现的纤维细且染色较淡,扫描电镜下神经元胞形较小,且大小不等,密较较小,神经元间的纤维联系及神经纤维攀附于肌纤维表面的现象均较少,神经元和神经纤维呈沿纵行肌长轴线性分布。结论：先天性巨结肠病的发生及发展可与NOS阳性神经元在肠壁的分布与代谢异常有关关。     

大鼠舌内NOS阳性神经元的分布及NOS与AChE共存的研究

本文用NADPH－d组化法和AChE显示法对大鼠舌内NOS阳性结构的正常分布以及NOS与AChE的共存进行了观察。结果表明，含NOS阳性神经元分布于舌的肌层、结缔组织、血管和味腺周围，其中NOS阳性细胞体以舌体游离段和舌根分布较多，舌尖和香体附着段次之，它们发出的神经纤维主要支配血管、腺体和肌组织．NOS和AChE双重染色表明两者分布模式一致。上述分布特征说明，NOS阳性神经元可能主要与血流调节和腺分泌有关，并参与胆碱能神经传递。     

扬子鳄胸髓一氧化氮合酶和乙酰胆碱酯酶阳性神经元的分布

目的观察一氧化氮合酶(NOS)和乙酰胆碱酯酶(AChE)阳性神经元在扬子鳄胸髓的分布。方法采用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)法和亚铁氰化铜法观察扬子鳄胸髓NOS和AChE阳性神经元的分布。结果扬子鳄胸髓前角、后角和中央灰质内可见NOS和AChE阳性神经元,白质内含有丰富的NOS和AChE阳性神经纤维。结论扬子鳄胸髓有NOS和AChE阳性神经元分布。     

正常成年大鼠脊髓灰质nNOS阳性神经元的形态学探察

目的探察大鼠脊髓灰质n NOS阳性神经元的形态和分布特征。方法免疫组织化学PAP方法被利用去探察大鼠颈、胸和腰骶脊髓灰质不同区域n NOS阳性神经元的大小、数量及分布。结果光镜观察显示,n NOS阳性神经元明显集中分布于脊髓中央管外周的灰质(中间带),但在腰骶脊髓灰质背侧角Ⅱ、Ⅲ层也呈现密集分布,而在颈脊髓和胸脊髓灰质Ⅳ和Ⅴ层同时可见散在分布。比较结果显示,腰骶脊髓灰质中间带的n NOS阳性神经元显著多于颈脊髓和胸脊髓(P0.05),而且n NOS阳性神经元在颈、胸和腰骶脊髓灰质的中间带均多于它们各自的背侧角(P0.05、P0.05、P0.05)。进一步对n NOS阳性神经元大小的比较结果显示,腰骶脊髓灰质中间带的n NOS阳性神经元明显大于颈脊髓和胸脊髓(P0.05),但阳性胞体的大小在颈、胸、腰骶脊髓灰质背侧角相互之间无统计学差异(P0.05)。结论大鼠脊髓灰质n NOS阳性神经元形态和分布特征可能牵涉到脊髓不同部位和区域的机能。     

先天性巨结肠肠壁NOS阳性神经元定性定量研究

目的:探讨一氧化氮合酶(NOS)阳性神经元与先天性巨结肠发生的关系。方法:对扩张段和移行段分别在双目解剖显微镜下做全层铺片,以NADPH-d酶组化法着色,光学显微镜下观察并以医学图像测算软件对神经元的面积、光密度等作量化处理。结果:扩张段神经节和神经纤维排列层次明显,色深量多,节内神经元色深,沿神经节周边及突起的基部排列。移行段神经节及神经纤维杂乱,量少色浅,节内神经元多浅染或未染,着色程度有较大差异。移行段近端、中段、远端神经元着色依次减弱。结论:NOS阳性神经元分布与代谢异常使肠壁舒张不能,部分导致了先天性巨结肠的发生。     

小鼠心脏一氧化氮合酶神经纤维的分布

目的：探讨小鼠心脏一氧化氮合酶aninic oxide synthse神经元结构和神经纤维分布。方法：NADPH-d组织化学技术。结果：小鼠心脏神经元主要分为NOS强阳性、中度和弱阳性反应细胞；心脏各部均接受NOS神经支配，其神经纤维多与肌纤维长轴平行走向。心房最丰富，房室结次之，左、右心室最少。心房及房室结的NOS阳性纤维呈串珠状膨大，心室的常为丝状，膨体极少。结论：小鼠心脏NOS神经元包括强阳性、中度和弱阳性反应细胞，NOS阳性纤维在心脏各部的分布和形态均有差异，NO可能作为神经递质和/或神经调节剂在心血流和冲动传导等的神经调控中起作用。     

人空肠NPY能神经分布的研究

本文用免疫组织化学ABC法,对神经肽Y(NPY)能神经纤维和神经元在人空肠壁内的分布进行了研究.结果:NPY能神经纤维和神经元呈棕褐色;NPY能神经纤维遍布肠壁各层,位于粘膜固有层内的神经纤维在小肠腺周围交织或疏网状.NPY能神经元见于肌间及粘膜下神经丛,尤以后者为多.粘膜内也可见NPY能神经元,它们分布于小肠腺下方,紧靠粘膜肌,或位于粘膜肌内.     

一氧化氮合酶阳性神经元在肝硬化大鼠中缝核的分布

目的研究肝硬化大鼠中缝核内一氧化氮合酶阳性神经元分布的变化。方法用四氯化碳建立肝硬化动物模型,NADPH-d组织化学方法观察大鼠脑干中缝核内神经元的变化,图像分析仪对神经元的形态及数量进行分析。结果①NOS阳性神经元及纤维广泛分布于脑干中缝核内,特别是中缝背核,且上段多于下段;②肝硬化大鼠NOS阳性神经元脑干中缝核内分布与正常组基本一致,但阳性反应物的密集度明显增高。结论肝硬化后脑干中缝核一氧化氮合酶较正常明显增多,其分布与兴奋性氨基酸分布相互重叠,可以推测它与一些神经递质在脑内共存并激发神经毒作用。     

P物质、血管活性肠肽和乙酰胆碱能神经在大鼠肠道内的分布及其关系

应用乙酰胆碱酯酶(AChE)组织化学和PAP免疫组织化学方法,比较观察P物质(SP)、血管活性肠肽(VIP)和AChE三种阳性神经元在大鼠十二指肠、空肠、回肠、结肠和直肠内的分布特征及其相互关系。结果显示:SP、VIP、AChE阳性神经神经元和纤维均分布于肠壁各层,从十二指肠、空肠到回肠逐渐增多,但从结肠到直肠则逐渐减少;AChE阳性神经元或纤维在肠壁各层最丰富,其中VIP以粘膜层和粘膜下神经丛较丰富,SP以肠肌丛较丰富;三者的分布密度为AChE>VIP>SP。AChE、SP和VIP阳性神经元胞体及神经纤维在不同肠段的分布密度有明显差异(P<0.05),提示可能与不同肠段肠动力调节功能有关。     

Colocalization of nitric oxide synthase and NADPH-diaphorase in the myenteric plexus of the rat gut.

The pattern of distribution and colocalization of nitric oxide-synthase (NOS) and NADPH-diaphorase in the myenteric plexus of whole-mount preparations of the antrum, duodenum, ileum, caecum, proximal colon and distal colon of the rat were investigated using immunohistochemical and histochemical staining techniques. Almost all the myenteric neurons that were NOS-positive in all regions of the gut examined were also stained for NADPH-diaphorase. However, in the stomach, duodenum and ileum, only a few of the NOS-positive nerve fibres in the tertiary and secondary plexuses and circular muscle layer were also stained for NADPH-diaphorase, whereas in the caecum and distal colon almost all the NOS-positive nerve fibres were also stained for NADPH-diaphorase. The results in the present study are consistent with the view that nitric oxide (NO) has a mediating role in gastrointestinal neurotransmission.     

Nitric oxide synthase and GABA colocalize in lamina II of rat spinal cord.

A dense plexus of fibers in the substantia gelatinosa contains nitric oxide synthase (NOS). Using electron microscopic double-labeling immunocytochemistry for NOS combined with GABA or glutamate, we find that all NOS-positive terminals in this region also contain GABA but are not enriched in glutamate. In an attempt to verify that NOS-positive terminals do not originate from primary afferents, we combined NOS immunocytochemistry with anterograde tracing from the sciatic nerve. An intrinsic spinal origin for the NOS-positive plexus is suggested. The results are discussed in the light of the possible involvement of nitric oxide in hyperalgesia.     

The neural regulation of the mammalian esophageal motility and its implication for esophageal diseases

In contrast to the tunica muscularis of the stomach, small intestine and large intestine, the external muscle layer of the mammalian esophagus contains not only smooth muscle but also striated muscle fibers. Although the swallowing pattern generator initiates the peristaltic movement via vagal preganglionic neurons that project to the myenteric ganglia in the smooth muscle esophagus, the progressing front of contraction is organized by a local reflex circuit composed by intrinsic neurons similarly to other gastrointestinal tracts. On the other hand, the peristalsis of the striated muscle esophagus is both initiated and organized by the swallowing pattern generator via vagal motor neurons that directly innervate the muscle fibers. The presence of a distinct ganglionated myenteric plexus in the striated muscle portion of the esophagus had been enigmatic and neglected in terms of peristaltic control for a long time. Recently, the regulatory roles of intrinsic neurons in the esophageal striated muscle have been clarified. It was reported that esophageal striated muscle receives dual innervation from both vagal motor fibers originating in the brainstem and varicose intrinsic nerve fibers originating in the myenteric plexus, which is called ‘enteric co-innervation’ of esophageal motor endplates. Moreover, a putative local neural reflex pathway that can control the motility of the striated muscle was identified in the rodent esophagus. This reflex circuit consists of primary afferent neurons and myenteric neurons, which can modulate the release of neurotransmitters from vagal motor neurons in the striated muscle esophagus. The pathogenesis of some esophageal disorders such as achalasia and gastroesophageal reflux disease might be involved in dysfunction of the neural networks including alterations of the myenteric neurons. These evidences indicate the physiological and pathological significance of intrinsic nervous system in the regulation of the esophageal motility. In addition, it is assumed that the components of intrinsic neurons might be therapeutic targets for several esophageal diseases.     

Somatostatin immunopositive neurons in the small intestine of the bullfrog (Rana catesbeiana).

Somatostatin immunopositive neurons in the small intestine of the bullfrog (Rana catesbeiana) were studied using immunohistochemistry and surgical denervation of the mesenteric nerve. Immunopositive nerve elements were distributed throughout the small intestine, including nerve fibers in the myenteric plexus, circular muscle layer, submucosal layer, and mucosa. Somatostatin immunopositive nerve cell bodies occurred in the myenteric plexus but not in the submucosal layer. These cell bodies were surrounded by immunopositive nerve fibers forming basket-like terminals, and thus some of these cells may be interneurons. After denervation of the mesenteric nerve, adrenaline immunopositive nerve fibers disappeared almost completely from the small intestine, but no changes occurred in the distribution of somatostatin immunopositive neurons. Neurons in the coeliac ganglion projecting into the small intestine were adrenaline immunopositive but somatostatin immunonegative. The results indicate that somatostatin immunopositive neurons in the small intestine of the bullfrog are primarily intrinsic in origin.     

一氧化氮合酶在大鼠主盆神经节及阴茎勃起组织中的分布

用ＮＡＤＰＨ脱氢酶组织化学方法观察了大鼠主盆神经节及阴茎勃起组织内一氧化氮合酶阳性成分的分布，发现主盆神经节内分布有大量ＮＯＳ阳性神经元，其中大部分密集于盆神经进入端，而膀胱端稀少，阴茎深动脉及其分支螺旋动脉周围，阴茎和尿道海绵体的平滑肌小梁内均分布有ＮＯＳ阳性纤维，阴茎勃起组织内未见ＮＯＳ阳性神经元，将光金注入阴茎海绵体后，在主盆神经节内发现有较多的荧光金标记细胞，结构ＮＡＤＰＨ反应，将荧光金注     

局部皮质内注射海人酸对皮质和海马一氧化氮合酶阳性神经元的影响

本实验用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶组织化学方法结合ＧＦＡＰ免疫组化方法，对微量海人酸皮质内注射后的一氧化氮合酶阳性神经元和胶质细胞等的变化进行了研究．结果发现：注射区内的一氧化氮合欧阳性神经元很快消失；注射侧皮质和海马的旧阳性神经元和神经末梢溃变；至注射海人酸后８ｈ，这些变化波及到对侧皮质和海马．随着皮质内一氧化氮合酶阳性神经元的溃变，在注射侧皮质和海马内的一些神经元、星形胶质细胞出现新的一氧化氮合酶活性，这些新的酶活性可能是诱导型的一氧化氮合酶．这些结果表明：表达一氧化氮合酶的皮质和海马神经元对海人酸的毒性很敏感，神经细胞和非神经细胞中出现的诱导型的一氧化氮合酶可能是海人酸引起脑损伤时机体的适应性反应．     

Acetylcholinesterase and choline acetyltransferase staining of neurons in the opossum esophagus

The purpose of the present investigation was to identify and compare cholinergic intramural neurons in the lower esophageal sphincter and esophageal body by histochemical staining for acetylcholinesterase and the enzyme that synthesizes acetylcholine, choline acetyltransferase. Opossums were anesthetized and their abdominal cavity was opened by a midline incision to expose the esophagogastric junction. The lower esophageal sphincter was identified manometerically and localized in situ with markers. Tissues were removed, rapidly frozen in freon cooled with liquid nitrogen and serial cryostat sections were obtained from the lower esophageal sphincter and esophageal body. Sections were stained with one of the above histochemical procedures and adjacent sections were stained with Solachrome cyanin , which differentially stains nerve elements from muscle fibers. The muscle of the lower esophageal sphincter and esophageal body was stained with nonspecific cholinesterase with some selectivity of intensity of reaction in the various smooth muscle layers. All identifiable plexus neurons in the esophagus stained for nonspecific cholinesterase and acetylcholinesterase. Nerve fiber tracts were also stained for acetylcholinesterase within the longitudinal and circular layers of the tunica muscularis. Reaction for choline acetyltransferase showed no staining in the muscle layers or nerve fiber tracts of either part of the esophagus studied; however, selected neurons within the myenteric plexus of both regions (approximately 38%) were reactive. There was no significant difference in the number of positive choline acetyltransferase neurons in the lower esophageal sphincter or esophageal body.     

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.