大鼠食管肌间神经元nNOS与calbindin的共存及其与衰老关系的研究(英文)

本实验通过观察一氧化氮合酶(nNOS)与细胞内钙结合蛋白(calbindin, CB)共存情况,旨在研究大鼠食管内NO类神经元的神经化学特性并试图寻找出引起食管NO类神经元在衰老过程中丢失的可能原因。取自不同年龄组及种系(Wistar和S-D)大鼠的食管组织,制成肌间神经丛铺片,经nNOS与CB免疫组化双重染色后在荧光显微镜下观察。结果显示,nNOS与CB的免疫阳性反应物均见于大鼠食管肌间神经元胞体内。根据二者共存情况,大鼠食管内nNOS免疫阳性神经元可分为两大亚类:nNOS+/CB+及nNOS+/CB-。而且我们首次发现在大鼠食管腹腔段含有大量nNOS+/CB+神经元,约占NO类神经元总体的30% ~40%,与食管其它部位相比有显著差异(P<0.001)。另外,大鼠食管腹腔段绝大多数CB免疫阳性神经元同时也为nNOS免疫阳性(约占95% ~100%)。上述特征均见于本实验所观察的所有不同年龄及不同种系的大鼠食管内。在大鼠衰老过程中,nNOS+/CB+神经元的相对百分比的变化具有种系特异性。线形回归实验分析表明:年轻SD大鼠食管内nNOS+/CB+神经元的百分比含量与衰老过程中NO类神经元丢失数量的百分比之间存在显著负相关性(correlation coefficient 0.99;P<0.05)。结果提示在S-D大鼠衰老过程中其食管内NO类神经元的丢失可能与其细胞内钙结合蛋白含量的变化有关。但nNOS/CB共存神经元在分布上有明显差异的生物学意义尚有待于进一步研究。     

大鼠舌组织中肥大细胞与肽能神经纤维的分布

目的探讨舌组织内肥大细胞与肽能神经的关系及其功能意义。方法采用甲苯胺蓝染色和免疫组织化学ABC染色法对10只Wistar大鼠舌组织进行SP、VIP和NPY染色。结果经与甲苯胺蓝邻片对比观察,舌组织内肥大细胞分别呈SP、VIP和NPY免疫反应性;在舌组织固有层及肌间结缔组织中还分布有丰富的SP、VIP和NPY免疫反应阳性纤维;在肽能神经纤维附近或周围可见有免疫反应阳性的肥大细胞,这些肥大细胞与神经纤维紧密相靠或接触。结论 大鼠舌组织内肥大细胞和神经纤维分别呈SP、VIP和NPY免疫反应性,提示肥大细胞与周围神经无论是在形态还是在功能上有着密切的联系。     

食管鳞癌组织中的肽能神经支配研究

目的　探讨食管鳞癌组织中的肽能神经支配情况。　方法　采用免疫组织化学ABC法 ,检查外科手术取得的食管鳞癌标本中 10种神经肽免疫反应性神经纤维的分布及其与肿瘤细胞的关系 ;采用食管鳞癌组织块和鸡胚背根神经节共培养的方法 ,初步探讨肿瘤组织对神经元的影响 ,以推测肿瘤中神经纤维的来源。　结果　食管鳞癌组织中存在相当数量的GAL、SP、NPY等多种神经肽免疫反应神经纤维 ,神经纤维与肿瘤细胞紧密接触。共培养组有约 6 3%的神经节长出突起 ,其中有 49%的神经节突起在靠近肿瘤组织块的一侧较浓密 ,对侧则较稀少。对照组鸡胚神经节则无突起生长。　结论　食管鳞癌组织中可能存在肽能神经支配 ;食管鳞癌组织块对神经节突起具有促生长作用。     

大鼠迷走神经初级传入纤维向孤束核投射的定位、定性研究

为了探索迷走神经向孤束核(NTS)投射纤维的性质及分布特征,本文应用(1) 免疫细胞化学技术对NTS及结状神经节(NG)内ENK、SP、CGRP和VIP等四种活性物质的存在及分布进行了普查;(2) 用HRP跨越神经节追踪技术探索了迷走神经初级传入纤维在NTS内的投射定位;(3) 用HRP逆行标记技术和免疫细胞化学双重标记法结合切断实验印证了NG内神经元向NTS投射的纤维性质。结果证明:NST内含有此四种活性物质的阳性纤维和终末,主要分布于连合核及NTS内侧部,即相当于内侧亚核、中间亚核的区域。NTS中段的尾侧及尾段的吻侧阳性纤维和终末密度最高。此四种活性物质以SP样和L-ENK样阳性成分最为密集,VIP样次之,CGRP样密度最低。NG内亦存在含此四种活性物质的阳性神经元,其中L-ENK样阳性神经元是本文首次发现的,跨节追踪证明的迷走神经初级传入纤维向NTS的投射部位与这四种性质的纤维扣终末密集区大体上一致。逆行追踪与免疫细胞化学相结合的结果证明NG内存在此四种性质的双重标记细胞。切断一侧NG中枢端发现在术侧NTS内此四种活性物质的阳性纤维和终末的密集区都出现“脱落现象”,说明NG内含此四种活性物质的神经元都向NTS投射。     

大鼠上涎核的传出神经纤维与翼腭神经节中的VIP-、DβH-、NPY-神经元的关系──光镜研究

目的　探讨翼腭神经节节前神经纤维在翼腭神经节中的形态、分布 ,以及与含不同神经活性物质的神经节细胞的关系。　方法　用顺行标记结合免疫组织化学方法进行研究。　结果　在翼腭神经节内 ,有大量的顺行追踪标记阳性神经纤维 ,呈篮状缠绕在神经节细胞周围 ,非常密集。免疫双标记法显示这些被顺行追踪阳性神经纤维包绕的神经元多呈 VIP、DβH和 NPY免疫反应性。在顺行追踪免疫反应性篮状神经纤维之间 ,还有 SP、CGRP、VIP、DβH、NPY免疫反应性神经纤维。　结论　翼腭神经节中 VIP(Ch AT)和 DβH、VIP免疫反应性神经元受到上涎核发出的节前神经纤维的调控     

大鼠上涎核的传出神经纤维与翼腭神经节中的VIP-、DβH-、NPY-神经元的关系光镜研究

目的探讨翼腭神经节节前神经纤维在翼腭神经节中的形态、分布,以及与含不同神经活性物质的神经节细胞的关系. 方法用顺行标记结合免疫组织化学方法进行研究. 结果在翼腭神经节内,有大量的顺行追踪标记阳性神经纤维,呈篮状缠绕在神经节细胞周围,非常密集.免疫双标记法显示这些被顺行追踪阳性神经纤维包绕的神经元多呈VIP、DβH和NPY免疫反应性.在顺行追踪免疫反应性篮状神经纤维之间,还有SP、CGRP、VIP、DβH、NPY免疫反应性神经纤维. 结论翼腭神经节中VIP(ChAT)和DβH、VIP免疫反应性神经元受到上涎核发出的节前神经纤维的调控.     

犬心脏表面主要神经丛降钙素基因相关肽和P物质的免疫组织化学研究

目的 探明犬心脏表面神经丛的化学特性。方法 免疫组织化学ABC法。结果 在犬心脏表面各神经丛均见降钙素基因相关肽(CGRP)免疫反应阳性神经元，而SP免疫阳性神经元仅在心房背侧神经丛(DAP)、房间隔神经丛(IAP)和主动脉．肺动脉间神经丛(A—PP)内见到。CGRP—IR和SP—IR神经元形态、大小相似。心房表面神经丛内的CGRP—IR和SP—IR神经元都较心室表面神经丛者多。在心脏表面各脂肪垫及心肌间隙等处见到多量CGRP-IR、SP—IR神经纤维，多靠近血管或附于血管壁，在一些部位可见这两种肽能神经纤维似与心肌细胞接触。结论犬心脏表面神经丛内存在CGRP和SP；其在心脏内执行的功能可能有联系或相似之处，但也有不同；两种肽能神经对心房和心室的支配不对称，提示CGRP和SP可能直接参与心肌细胞和心脏血管活动的调控。     

炎性胃痛幽门括约肌内VIP和CGRP能神经的变化

目的：研究大鼠幽门括约肌内肠神经系统的分布与胃肠运动功能的相互关系。方法：30只大鼠，设对照组，实验组，采用免疫组织化学技术，显示括约肌内VIP－和CGRP－ir神经元成分，结果：（1）正常幽门括约肌肌层可见个别的CGRP－和VIP－ir神经元胞体，阳性纤维丰富，于肌纤维增厚的括约肌部位神经元成分特别丰富，（2）甲醛致胃炎性痛鼠，肌间神经丛VIP神经元胞体和肌层内神经纤维的活性（图像分析，平均光密度）降低，甲醛组比对照组均显著降低（P＜0．01），但CGRP的甲醛组较对照组升高（P＜0．01），结论：肠神经系统中这两种神经元成分参与幽门括约肌活动与胃肠运动的调控机制。     

大鼠三叉神经脊束核尾侧亚核及三叉神经节内PPTA、CGRP和PPEmRNA及其相应蛋白在面口部炎性刺激时的变化

利用原位杂交技术和免疫组化技术，观察完全福氏佐剂致炎大鼠的三叉神经脊柬核尾侧亚核和／或三叉神经节内SP、CGRP和L-ENK样阳性结构及它们相应的前体PPTAmRNA、CGRPmRNA和PPEmRNA分布状况的动态变化。原位杂交结果显示：PPTAmRNA在三叉神经节与三叉神经脊束核尾侧亚核浅层神经元中的表达于致炎刺激后12h明显增强，分别于1d和4d时达到高峰，21d后恢复正常;CGRPmRNA在三叉神经节神经元中的表达变化与PPTAmRNA者相似;在三叉神经脊束核尾侧亚核浅层PPEmRNA阳性神经元的数量于刺激后12d明显增多，8d到达高速，28d时仍高于正常水平.免疫组织化学结果显示：尾侧亚核浅层SP样阳性纤维和终末的密度在致炎刺激后12h至4d降低，然后逐渐回升到正常水平;而CGRP样阳性纤维和终末的密度在佐剂注射后1d明显降低，8d后恢复正常，14d后又明显高于正常水平;佐剂刺激后1d，L-ENK样阳性纤维和终末的密度明显增加，以后维持正常水平，21d和28d时又低于正常水平.在三叉神经节，SP和CGRP样阳性神经元的数量在致炎刺激后早期明显减少，以后恢复至正常水平且再无明显变化。上述结果表明，在完全福氏佐剂的致炎条件下，SP、CGRP和L-ENK样阳性结构及其相应的mRNA在尾侧亚核浅层和／或三叉神经节存在着不同的时程变化，反映了它们在     

5-羟色胺3A受体在成年小鼠海马中间神经元中的分布

目的:利用成年5-HT3AR-BACEGFP转基因小鼠,研究5-羟色胺3A受体(5-HT3AR)在海马中间神经元中的分布情况。方法:成年5-HT3AR-BACEGFP转基因小鼠经心脏灌注固定后,利用免疫荧光双标记方法,并结合激光共焦显微镜扫描技术,观察5-HT3AR在成年5-HT3AR-BACEGFP转基因小鼠海马中不同中间神经元内的表达和分布情况。结果:5-HT3AR在成年小鼠整个海马中都有分布,且主要在CA1区、CA2/CA3区和齿状回有大量5-HT3AR免疫阳性细胞;激光共聚焦显微镜下观察到5-HT3AR阳性产物在细胞核、细胞浆和树突上均有表达;免疫荧光双标实验结果表明5-HT3AR阳性产物在CB(calbindin),CR(calretinin),Reelin,Som(somatostatin),NPY(neuropeptide Y)和VIP(vasoactive intestinal peptide)免疫阳性神经元中表达,但在PV(parvalbumin)免疫阳性神经元中不表达。定量结果显示:几乎所有的VIP阳性神经元均表达5-HT3AR阳性,约3/4的CR阳性神经元表达5-HT3AR,约1/2的CB、Reelin、NPY和Som阳性神经元表达5-HT3AR阳性;约1/4的5-HT3AR阳性神经元中表达Reelin,1/5的表达Som,5-HT3AR/CB和5-HT3AR/CR双标神经元各占5-HT3AR阳性神经元的1/10左右。结论:5-HT3AR-BACEGFP转基因小鼠能够作为研究海马中5-HT3AR功能及其在中间神经元中的作用机制研究的工具鼠。     

Light and electron microscopic studies of pituitary adenylate cyclase-activating peptide (PACAP) – immunoreactive neurons in the enteric nervous system of rat small and large intestine

 Pituitary adenylate cyclase-activating peptide (PACAP)-immunoreactive (IR) neurons in the myenteric and submucosal plexus of the rat small and large intestine were examined by immunostaining with purified polyclonal antiserum against PACAP (1–15), using both light and electron microscopy. Many PACAP-IR neuronal cell bodies and fibers were found in the myenteric and submucosal plexus. Many of the PACAP-IR fibers originated from the cell bodies of the myenteric and submucosal ganglia. The ganglia were also innervated by PACAP-IR fibers. PACAP-IR fibers penetrated both the circular and longitudinal muscle layers, confirming the previous observations indicating that PACAP neurons act as motor neurons. Ultrastructural study demonstrated that PACAP-IR nerve terminals formed synaptic contacts with PACAP-IR nerve cell bodies or dendritic processes. This observation suggests that PACAP-IR neurons innervate other PACAP-IR neurons, and that PACAP neurons work as interneurons in the enteric nervous system. PACAP-IR nerve cells received not only PACAP-positive nerve terminal input also PACAP-negative nerve terminal input. It also suggests that PACAP neurons are regulated not only by PACAP-IR enteric neurons, but also by neurons originating elsewhere. Our observations support the view that PACAP-IR neurons are involved in the control of gut motility. Accepted: 20 April 1998     

Innervation pattern of polycystic ovaries in the women

The aim of the present study was to determine the changes in both the distribution pattern and density of nerve fibers containing dopamine β-hydroxylase (DβH), vesicular acetylcholine transporter (VAChT), neuronal nitric oxide synthase (nNOS), substance P (SP), calcitonin gene related peptide (CGRP), neuropeptide Y (NPY), vasoactive intestinal peptide (VIP), somatostatin (SOM), galanin (GAL) and pituitary adenylate cyclase-activating polypeptide (PACAP) in the human polycystic ovaries. In the polycystic ovaries, when compared to the immunoreactions pattern observed in the control gonads, following changes were revealed: (1) an increase in the number of DβH-, VAChT-, VIP- or GAL-immunoreactive (IR) nerve fibers within the stroma as well as in the number of DβH-IR fibers near primordial follicles and medullar veins and venules; (2) a reduction in the number of nerve fibers containing nNOS, CGRP, SOM, PACAP within the stroma and in the numbers of CGRP-IR fibers around arteries; (3) an appearance of SP- and GAL-IR fibers around medullar and cortical arteries, arterioles, veins and venules, with except of GAL-IR fibers supplying medullar veins; and (4) the lack of nNOS-IR nerve fibers near primordial follicles and VIP-IR nerves around medullar arteries and arterioles. In conclusion, our results suggest that the changes in the innervation pattern of the polycystic ovaries in human may play an important role in the pathogenesis and/or course of this disorder.     

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.     

Ontogeny of the VIP system in the gastro-intestinal tract of the Axolotl,<Emphasis Type="Italic"> Ambystoma mexicanum</Emphasis>: successive appearance of co-existing PACAP and NOS

Evidence for the presence and potential co-existence of vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating polypeptide (PACAP) and nitric oxide synthase (NOS) in gastro-intestinal endocrine cells and/or nerve fibers is conflicting and very few results exist on development. This immunofluorescence study aims to clarify the appearance and localization of VIP, PACAP and NOS in the gastro-intestinal tract of the Axolotl, Ambystoma mexicanum, during ontogeny. VIP-immunoreactivity appeared in nerve fibers as early as on day 3 after hatching likely indicating a particular role, such as a trophic action, of VIP in very early development. PACAP-immunoreactivity was observed 3 days later within the VIP-immunoreactive (-IR) fibers. From this time on, VIP- and PACAP-immunoreactivity exhibited complete co-existence. VIP/PACAP-IR fibers were found throughout the gastro-intestinal tract. They were most prominent in the myenteric plexus and the muscle layers and less frequent in the submucosa. NOS-immunoreactivity appeared as late as at the 1st (64 days) juvenile stage in a subpopulation of the VIP/PACAP-IR fibers that contacted submucosal arteries. We found only very few VIP/PACAP-IR perikarya, indicating that part of the VIP/PACAP-IR fibers is of extrinsic origin. On day 12 and in the 1st and 2nd (104 days) juvenile stage, infrequent PACAP-IR entero-endocrine cells were noted, while neither VIP- nor NOS-immunoreactivity occurred in endocrine cells at any stage of development. The complete coexistence of neuronal PACAP- and VIP-immunoreactivities and their very early appearance in ontogeny may suggest important and coordinated roles of both peptides in the control of Axolotl gastro-intestinal activity, while the VIP/ PACAP/NOS-IR fibers may be involved in the regulation of submucosal blood flow.     

Immunocytochemical analysis of potential neurotransmitters present in the myenteric plexus and muscular layers of the corpus of the guinea pig stomach

Recent electrophysiological studies of neurons of the myenteric plexus of the corpus of the guinea pig stomach have revealed that slow synaptic events are extremely rare. In contrast, they are commonly encountered in similar investigations of myenteric ganglia of the guinea pig small intestine. The current immunocytochemical analysis of the myenteric plexus and innervation of the muscularis externa of the corpus of the guinea pig stomach was undertaken in order to determine whether putative neurotransmitters capable of mediating slow synaptic events are present in gastric ganglia. A major difference between the small intestine and the stomach was found in the innervation of the musculature. Whereas the longitudinal muscle layer of the small intestine contains very few nerve fibers and is innervated mainly at its interface with the myenteric plexus, the longitudinal muscle of the corpus of the stomach contained as many varicose substance P (SP)-, vasocative intestinal polypeptide (VIP)-, and neuropeptide Y (NPY)-immunoreactive axons as the circular muscle layer. These putative neurotransmitters were also present in the ganglia of the myenteric plexus, where varicose SP-, VIP-, and NPY-immunoreactive fibers encircled nonimmunoreactive neurons. Varicose 5-hydroxytryptamine (5-HT)-immunoreactive terminal axons were essentially limited to the myenteric plexus and were found both in ganglia and in interganglionic connectives, where they were particularly numerous; 5-HT-immunoreactive neurons appeared to be more abundant in the stomach than in the small intestine. Tyrosine hydroxylase (TH)- and calcitonin-gene-related-peptide (CGRP)-immunoreactive axons were also more common in the myenteric plexus than in the musculature, but of these, only the TH-immunoreactive neurites tended, like those of the other putative transmitters, to encircle neurons in myenteric ganglia. Evidence was obtained that, as in the small intestine, at least some of the SP-, VIP-, NPY-, and 5-HT-immunoreactive fibers in the stomach are derived from intrinsic gastric myenteric neurons. In contrast, unlike the small intestine, gastric myenteric ganglia appeared to lack intrinsic CGRP-immunoreactive neurons; therefore, the CGRP-immunoreactive gastric axons are probably of extrinsic origin.(ABSTRACT TRUNCATED AT 400 WORDS)     

Nocturnal motor coordination deficits in neuronal nitric oxide synthase knock-out mice

Nitric oxide is formed in the brain primarily by neurons containing neuronal nitric oxide synthase (nNOS), though some neurons may express endothelial NOS (eNOS), and inducible NOS (iNOS) only occurs in neurons following toxic stimuli. Mice with targeted disruption of nNOS (nNOS-) display distended stomachs with hypertrophied pyloric sphincters reflecting loss of nNOS in myenteric plexus neurons. nNOS- animals resist brain damage following middle cerebral artery occlusions consistent with evidence that excess release of nitric oxide mediates neurotoxicity in ischemic stroke. Neuronal NOS- mice have no grossly evident defects in locomotor activity, breeding long-term depression in the cerebellum, long-term potentiation in the hippocampus, and overall sensorimotor function. However, nNOS- animals display excessive, inappropriate sexual behavior and dramatic increases in aggression. Because the cerebellum possesses the greatest levels of nNOS neurons in the brain, it was surprising that presumed cerebellar functions such as balance and coordination were grossly normal in nNOS- mice. These previous studies were all conducted during the day (between 1400 and 1600, lights on at 0700). We now report striking, discrete abnormalities in balance and motor coordination in nNOS-mice reflected selectively at night.     

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.     

Ontogeny of the VIP system in the gastro-intestinal tract of the Axolotl,Ambystoma mexicanum: successive appearance of co-existing PACAP and NOS

Evidence for the presence and potential co-existence of vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating polypeptide (PACAP) and nitric oxide synthase (NOS) in gastro-intestinal endocrine cells and/or nerve fibers is conflicting and very few results exist on development. This immunofluorescence study aims to clarify the appearance and localization of VIP, PACAP and NOS in the gastro-intestinal tract of the Axolotl, Ambystoma mexicanum, during ontogeny. VIP-immunoreactivity appeared in nerve fibers as early as on day 3 after hatching likely indicating a particular role, such as a trophic action, of VIP in very early development. PACAP-immunoreactivity was observed 3 days later within the VIP-immunoreactive (-IR) fibers. From this time on, VIP- and PACAP-immunoreactivity exhibited complete co-existence. VIP/PACAP-IR fibers were found throughout the gastro-intestinal tract. They were most prominent in the myenteric plexus and the muscle layers and less frequent in the submucosa. NOS-immunoreactivity appeared as late as at the 1st (64 days) juvenile stage in a subpopulation of the VIP/PACAP-IR fibers that contacted submucosal arteries. We found only very few VIP/PACAP-IR perikarya, indicating that part of the VIP/PACAP-IR fibers is of extrinsic origin. On day 12 and in the 1st and 2nd (104 days) juvenile stage, infrequent PACAP-IR entero-endocrine cells were noted, while neither VIP- nor NOS-immunoreactivity occurred in endocrine cells at any stage of development. The complete coexistence of neuronal PACAP- and VIP-immunoreactivities and their very early appearance in ontogeny may suggest important and coordinated roles of both peptides in the control of Axolotl gastro-intestinal activity, while the VIP/ PACAP/NOS-IR fibers may be involved in the regulation of submucosal blood flow.     

迷走神经切断后大鼠心房室结区神经肽变化的研究

切断大鼠颈部一侧迷走神经后，用组织化学方法结合图像分析定量观察了心房室结区乙酰胆碱酯酶，神经肽Ｙ（ＮＰＹ）、血管活性肠肽（ＶＩＰ）和降钙素基因相关肽（ＣＧＲＰ）的变化，同时用生物深化法测定了ＡＶＮ区ＡＣＨＥ、去甲肾上腺素（ＮＡ）的含量。并用逐步回归分析了各递质间的相互关第。本实验发现：ＡＶＮ区有ＮＰＹ、ＶＩＰ、ＣＧＲＰ免疫反应神经纤维和神经细胞。图像分析表明，左侧或右侧迷走神经切断５ｄ后ＡＶＮ区Ｎ