与坐骨神经和盆神经对应的后根节中CGRP、SP、NOS样阳性神经元分布特点分析——荧光金逆标和荧光免疫组化结合的研究

为了探索大鼠坐骨神经（躯体性）和盆神经（内脏性）内与传递痛信号有关的初级传入神经元在后根节内的分布特点,本研究采用荧光金逆行追踪与免疫荧光组化技术相结合的方法,对ＣＧＲＰ能、ＳＰ 能和ＮＯＳ样神经元在相应的后根节内（坐骨神经,Ｌ４～Ｌ６ ;盆神经,Ｌ６～Ｓ１）的分布状况进行了分析。结果表明：（１）坐骨神经和盆神经初级传入神经元中有相当数量的ＣＧＲＰ和ＳＰ样阳性细胞,与这二者相比,ＮＯＳ样细胞数量稀少;（２）盆神经初级传入神经元中ＣＧＲＰ／ＦＧ、ＳＰ／ＦＧ、ＮＯＳ／ＦＧ 双标细胞的比率高于坐骨神经,而其前两种双标细胞与各该活性物质单标细胞的比率则低于坐骨神经;（３）三种物质与ＦＧ 的双标神经元以小型为主,少有中型细胞。因为既往的研究证明,分布有大量的ＣＧＲＰ、ＳＰ、ＮＯＳ样终末的骶髓后连合核（ＳＤＣＮ）接受盆腔脏器伤害性信息传入,并且ＣＧＲＰ、ＳＰ都以外周来源为主。故本文结果进一步核实了ＳＤＣＮ 区接受来自外周的ＣＧＲＰ、ＳＰ投射,且确为经盆神经传入的细纤维。     

I─B4阳性纤维和终末及P物质受体在大鼠腰骶髓内的分布

为了探讨传递伤害性刺激信息的Ｃ纤维终末与ＳＰ受体（ＳＰＲ）阳性结构间的关系，用Ｉ－Ｂ４组织化学法和ＳＰＲ免疫组织化学法，光镜下观察了大鼠腰骶髓内Ｉ－Ｂ４阳性纤维及其终末和ＳＰＲ阳性胞体及其树突的分布。结果显示：被Ｉ－Ｂ４特异标记的阳性纤维及其终末主要分布于腰骶髓后角Ｉ、ＩＩ层以及内、外侧束。另外，在腰骶髓的后连合核区（ＤＣＮ）也有相当数量的Ｉ－Ｂ４阳性纤维及其终末分布。说明了伤害性刺激信息主要传递至腰骶髓内的后角Ｉ、ＩＩ层，同时也有相当一部分伤害性刺激信息传递至腰骶髓的ＤＣＮ区。传递至ＤＣＮ区的Ｉ－Ｂ４阳性纤维主要通过沿后角内侧缘走行的内侧束，从而为ＤＣＮ区参与内脏伤害性刺激信息传递及其调制机制提供了进一步的证据。ＳＰＲ阳性胞体和树突主要分布于腰骶髓的后角Ｉ层、Ｉ～Ⅳ层，后角侧缘的内、外侧束也有少量分布。此外，ＤＣＮ区也有大量的ＳＰＲ阳性树突和散在的阳性胞体。双重反应的结果显示两者在Ｉ、ＩＩ层及ＤＣＮ区的分布相互匹配，从而推测被Ｉ－Ｂ４标记的初级传入Ｃ纤维及其终末可能含有Ｐ物质     

大鼠坐骨神经和盆神经初级传入纤维向腰骶髓后连合核的投射--HRP和BSI-B4-HRP跨越神经节追踪研究

目的　观察大鼠坐骨神经和盆神经初级传入纤维在腰骶段脊髓后连合核内的分布。　方法　应用HRP和BSI B4 HRP跨越神经节追踪技术。　结果　将HRP注射到坐骨神经后 ,有一定数量的HRP标记纤维出现于后连合核内 ,而BSI B4 HRP标记的坐骨神经初级传入纤维全部终止于后角浅层 (主要在Ⅱ层 ) ,后连合核内未见任何阳性标记。HRP标记的后根节神经元大、中、小均有 ,其平均直径为 33 2 5± 14 18μm ,而BSI B4 HRP标记的细胞以小型为主 ,其平均直径为 17 5 9± 4 80 μm。HRP和BSI B4 HRP标记的盆神经初级传入纤维在腰骶段脊髓内的分布相似 ,且均向后连合核投射 ,但BSI B4 HRP注入例的标记量明显少于HRP实验组。BSI B4 HRP标记的后根节神经元的数量也明显少于HRP实验组 ,但两者均以直径在 10～ 2 0 μm的小型细胞为主。 　结论　终止于后连合核的坐骨神经初级传入纤维可能为粗纤维 ,而盆神经则含有细纤维。这种躯体和内脏神经在后连合核内的不同终止形式可能与针刺镇痛的机制密切相关     

三叉初级感觉神经元的选择性标记--植物凝集素法光镜、电镜研究

为了探讨三叉初级传入路径中传导伤害性信息的小神经元在结构和功能上的差异，用植物凝集素（Ｂａｎｄｅｉｒａｅａｓｉｍｐｌｉｃｉｆｏｌｉａｉｓｏｌｅｃｔｉｎ－Ｂ４，ＢＳＩ－Ｂ４）法在光镜，电镜水平观察了三叉初级传入路径中标记神经元的分布及形态特征，光镜结构表明：ＢＳＩ－Ｂ４标记的初级传入终末只分布于三叉神经尾侧亚核的Ⅰ层和Ⅱ层，尤以Ⅱ层密集，标记终末光滑不呈串珠状。三叉神经节中的ＢＳＩ－Ｂ４标记神经为中     

骶髓后连合核内躯体初级传入粗纤维的证明及其意义探讨

近 2 0年来的研究表明 ,骶髓后连合核是接受和中继盆腔脏器初级传入信号的重要内脏感觉核团。通过盆神经传入脊髓的盆腔脏器初级传入神经纤维除一部分投射于中间带外侧核区参与排尿反射活动的调控外 ,其余部分基本上都投射于骶髓后连合核。本文作者等又曾发现躯体神经 (坐骨神经、阴部神经 )的初级传入粗纤维有一部分经后索投射于后连合核 ,推测这些躯体初级传入粗纤维可能与内脏传入细纤维汇聚于后连合核神经元并在此进行机能的整合、产生新的神经效应。但是投射于后连合区的躯体初级传入粗纤维 ,必须在电镜下直接证实。为此 ,本研究通过透射电镜及 HRP标记电镜技术探索了阴部神经躯体初级传入粗纤维在后连合核区的存在 ,并用 Philips CM-10 0电镜附件 Measuring装置测量了这种纤维的轴突直径和髓鞘厚度、通过计算求得了它们之间的比例 ( A/M值 ) ,藉此取得了科学地辨认并确证粗纤维的根据     

盆内脏感觉二级传人通路的研究

向猫盆神经注入10％蓖麻毒素2μl,存活3～5天后,再注射20％HRP溶液于同侧臂旁外侧核,2～3天后多聚甲醛和戊二醛灌注动物及取材。电镜下发现,盆神经初级传入纤维的终末(溃变)与向臂旁外侧核投射的位于骶髓后连合核、中间带外侧核、后角Ⅰ层内的HRP逆行标记神经元,形成轴-树和轴-体突触,从而在超微水平确证了盆内脏感觉传入的二级传导通路起源于骶髓后连合核、中间带外侧核、后角Ⅰ层,投射到臂旁外侧核。     

大鼠三叉神经脊束核尾侧亚核内SP受体阳性神经元、FOS阳性神经元及初级传入C纤维和终末分布的研究

用免疫组织化学和组织化学方法对大鼠三叉神经脊束核尾侧亚核（Ｖｃ）内ＳＰ受体阳性神经元、ＦＯＳ阳性神经元及初级传入Ｃ纤维和终末的分布及相互之间的关系进行了观察。ＳＰ受体阳性神经元的胞体及树突主要分布在此核的Ⅰ、Ⅱ层，Ⅰ层多于Ⅱ层；１０％福尔马林刺激大鼠口周区后诱导的ＦＯＳ阳性神经元分布于此核的Ⅰ～Ⅳ层及其内侧的网状结构，但主要集中在Ⅰ、Ⅱ层，Ⅱ层多于Ⅰ层；ＢＳＩ－Ｂ４标记的初级传入Ｃ纤维及其终末也密集地分布于此核的Ⅰ、Ⅱ层，且Ⅱ层多于Ⅰ层。多重反应的结果显示：在Ｖｃ的Ⅱ层和Ⅰ层内，ＳＰ受体阳性神经元的胞体及树突和ＦＯＳ阳性神经元胞体的周围可见许多ＢＳＩ－Ｂ４标记的纤维及终末；部分ＳＰ受体阳性神经元也呈ＦＯＳ阳性，少数ＳＰ受体和ＦＯＳ的双重阳性神经元周围也有ＢＳＩ－Ｂ４标记的纤维和终末。以上结果说明：（１）传递面口部痛信号的Ｃ纤维主要终止在尾侧亚核的Ⅰ、Ⅱ层，与ＳＰ受体阳性神经元的分布区重叠，提示这些Ｃ纤维可能含ＳＰ；（２）参与西口部痛信息传递的ＦＯＳ阳性神经元主要分布于Ｖｃ的Ⅱ层，它们与初级传入Ｃ纤维终末的分布区重叠，其中的部分ＰＯＳ阳性神经元也呈ＳＰ受体阳性，从而进一步地证明了ＢＳＩ－Ｂ４标记的初级传入Ｃ纤?     

I—B4阳性纤维和终末及P物质受体在大鼠腰骶髓内的分布

籽探讨传递伤害性刺激信息的Ｃ纤维张末与ＳＰ受体（ＳＰＲ）阳性结构间的关系，用Ｉ－Ｂ４组织化学法和ＳＰＲ免疫组织化学法，光镜下观察了大鼠腰骶髓内Ｉ－Ｂ４阳性纤维及其终末和ＳＰＲ阳性一胞体及其树突的分布、结果显示，被Ｉ－Ｂ４特异标记的阳性纤维及其终末主要分矶于腰骶后角Ⅰ、Ⅱ层以及内、外侧束。另餐；在腰骶髓的后连合核区（ＤＣＮ）也有相当数量的Ｉ－Ｂ４阳性纤维及其终末分布，说明了伤害性刺激信息主要传递至腰     

盆腔内脏感觉的二级传入通路——溃变和HRP标记光镜、电镜结合研究

<正> 关于盆腔内脏初级传入的行程和终止部位已有较多的研究,其结果认为主要终止于骶髓_(1-3)的Ⅰ、Ⅷ层及后连合核。史中立等研究证明,HRP注射于大鼠臂旁外侧核(PBL)在骶髓Ⅰ、Ⅷ层和后连合核可追踪到逆行标记细胞。因为PBL已被证明接受弧束核的内脏二级传入。本实验目的在于在电镜水平证实投射到PBL的骶髓Ⅰ、Ⅷ层和后连合核的神经元是否为盆腔内脏传入感觉二级神经元。家猫五只,将10％Ricin CA60 3μl注射一侧盆神经,3     

大鼠腰骶髓后连合核SP受体免疫阳性神经元与SP免疫阳性终末的突触联系

应用免疫组织化学双重反应的方法研究了大鼠腰骶髓后连合核内ＳＰ受体免疫阳性神经元在光、电镜下的特征及其与ＳＰ免疫阳性终末的突触联系。结果证明，ＳＰ免疫阳性终末和ＳＰ受体阳性胞体及树突均密集分布于后连合核区。在光镜下双重免疫反应阳性结果显示二者重叠分布，且可见大量的ＳＰ样免疫阳性终扣与ＳＰ受体阳性胞体或树突紧密接触。电镜下证明ＳＰ受体免疫阳性产物不仅位于突触后部位而且分布于胞体和树突的非突触部位，有４２％的ＳＰ受体阳性树突与ＳＰ阳性终末形成突触连接。在此未发现ＳＰ受体免疫阳性终末。通过一系列实验证明：腰骶髓后连合核神经元接受由ＳＰ介导的外周伤害性信息的传入，且其主要为盆腔脏器内脏初级传入成分。     

Morpholological analyses of galaninergic inputs to the rat spinal parasympathetic nucleus

We examined the characteristic features of galanin (GAL)-containing nerve afferents in the intermediolateral nucleus (IML) of the rat lumbosacral spinal cord (L6, S1), i.e., spinal parasympathetic nucleus, by immunocytochemistry at both light and electron microscopic levels. Firstly, the types of synapses formed by GAL-immunoreactive (IR) axon terminals and their post- or presynaptic elements were examined in random ultrathin sections. A total of 109 synapses were examined. Axo-dendritic (71%) and axo-somatic (20%) synapses were always of the asymmetrical type. Axo-axonic synapses (9%) were occasionally found; GAL-IR axon terminals were either postsynaptic (3%) or presynaptic (6%) to non-IR axon terminals. By confocal laser microscopy, many GAL-IR axon terminals were seen close to cell bodies and proximal dendrites of the IML neurons that were retrogradely labeled with Fluoro-Gold injected into the pelvic ganglion. Some GAL-IR axon terminals were identified to be presynaptic to them under the electron microscope, by restaining for GAL immunoreactivity with the immunoperoxidase method. These findings suggest that the GAL afferents are involved in the parasympathetic motor regulation of pelvic organs via their central synaptic influences upon preganglionic neurons. Finally, hemi-transection of the upper lumbar segments (L1-L3) or unilateral dorsal rhizotomy (L5-S2) did not significantly alter the immunoreactivity for GAL in the IML. These results suggest that GAL afferents do not originate from regions rostral to the IML nor from the dorsal root ganglion, but probably from GAL cells located at least within the lower lumbar segments and/or sacral spinal cord.     

Direct projections from the sacral spinal cord to the medial preoptic area in cat and guinea pig

The medial preoptic area (MPO) plays an important role in many behavioral, autonomic and endocrine functions, including micturition and genital responses. Although afferents of the MPO have been studied extensively, it is unknown whether direct lumbosacral-MPO projections exist that could convey afferent information from the pelvic organs. We hypothesized that, if such direct projections exist, MPO projecting cells would be located in the lateral part of the sacral cord, where primary afferents from pelvic and pudendal nerves terminate. We used retro- and anterograde tracing techniques in cat and guinea pig to study this. In cats, injections in the MPO resulted in labeled cells throughout the spinal cord, but with the highest density in the S1–S2 segments. In guinea pigs, labeled cells were found exclusively in the S1–S3 segments after MPO injections. Labeled cells in the sacral segments were not located in the lateral parts of the gray matter, but were found in the medial laminae VI–VII and dorsal lamina VIII in cats, and mainly in lamina X in guinea pigs. Anterograde tracing results after injections in the sacral cord in cats or guinea pigs showed labeled fibers in the MPO, just ventral to the anterior commissure. The central location of the cells of origin within the sacral cord, together with the termination pattern of the spino-MPO projections, strongly suggest a role for the spino-MPO pathway in the sensory relay of pelvic viscera, important for micturition and genital responses.     

猫骶髓“内脏面”传出神经元和二级传入神经元的分布

应用HRP和荧光金标记技术,在荧光显微镜下观察了猫骶體“內脏面”的传出神经元和二级传入神经元的分布?咀偌磷⑷肱枭窬?标记的骶髓副交感节前神经元主要分布于同侧中间带外侧核,由它组成的骶髓副交感核在以S2为中心的两个骶髓节段内(有时为三个),形成长约6-10mm的细胞柱。在横切面上,核的中段以上分为位于Ⅶ层外侧缘的外侧带和位于Ⅴ层的背侧带,但在核的尾侧段则二者融合为一。中介核和中间带内侧核也存在少量逆标神经元。示踪剂注于臂旁外侧核或Barrington核(一侧或双侧)后,逆标的盆内脏二级传入神经元主要存在于骶髓中间带外侧核和骶髓后连合核,在中间带外侧核位于两团传出神经元之间的中间带,但在核的尾侧段则位于骶髓副交感核的背外侧部,个别细胞混杂于副交感节前神经元之间,中介核和中间带內侧核也有少量二级传入神经元分布?送?相当数量的盆内脏二级传入神经元位于后角Ⅰ层。本文结果证明猫骶髓中间带外侧核至少由两种不同性质的神经元即骶髓副交感节前神经元和盆内脏二级传入神经元组成,两者在中间带外侧核区有明确的定位分布,因此,可将骶髓中间带外侧核划分为传出亚核和传入亚核,传统上笼统地将骶髓副交感核等同于中间带外侧核是不恰当的。     

犬马尾与骶神经根的解剖学观察

目的：为从马尾和骶神经根途径开展犬的神经泌尿学研究提供解剖依据。方法：对3只犬灌后进行解剖,并对22只犬进行术中观察,总结马尾、骶神经根的解剖特征。结果：犬的盆底器官由S1～S2脊髓节段和神经根支配;脊髓圆锥延续较长,达L6椎体下缘,而马尾神经较短;髓神经前后根出硬膜后,有各自独立的硬膜囊包绕直至后根神经节处,长1～1.5cm。结论：犬马尾和骶神经根的解剖特征与人类不同。     

Transganglionic transport of the lectin soybean agglutinin (Glycine max) following injection into the sciatic nerve of the adult rat

The lectin soybean agglutinin (SBA) from Glycine max binds to small-sized dorsal root ganglion cells and their central terminals in the superficial dorsal horn of the spinal cord. Here we investigated the ability of SBA and SBA conjugated to horseradish peroxidase(SBA-HRP) to trace thin calibre afferents into the spinal cord from a peripheral nerve. Following injection into the sciatic nerve, labelled cells in the dorsal root ganglion were predominantly small-sized but some medium-sized cells were also labelled. Colocalisation studies of transported SBA with various neuronal markers showed that all neurons that transported SBA-HRP showed SBA binding, indicating high uptake specificity for the conjugate. 15% were immunoreactive for RT97 indicating that some axons were myelinated, and 54% also expressed binding sites for isolectin B4 from Griffonia simplicifolia, a selective marker for a subpopulation of unmyelinated afferents. With regard to neurotransmitter content, 43% of the SBA cells contained calcitonin gene-related peptide, 33% contained substance P and 2.5% somatostatin. In addition, 3% contained carbonic anhydrase. Centrally, injection of SBA in the sciatic nerve resulted in labelled terminals in somatotopically appropriate regions of laminae I–II of the dorsal horn, and in the gracile nucleus. A few neurons in the dorsal horn were labelled indicating that some transneuronal transport of SBA had occurred. The results show that SBA can be used as a transganglionic tracer to label fine calibre primary afferents that project to laminae I–II of the spinal cord and the gracile nucleus. It appears to label more afferents than isolectin B4, including also a subpopulation of myelinated afferents.     

Electrical stimulation of visceral afferent pathways in the pelvic nerve increases c-fos in the rat lumbosacral spinal cord.

Electrical stimulation (20-35 Hz, 2-5 V, 1.5 h) of the pelvic nerve in urethane-anesthetized rats increased the expression of c-fos protein-immunoreactivity primarily in neurons in the L6-S1 segments of the spinal cord. The neurons were localized to areas receiving afferent input from the pelvic viscera including the superficial dorsal horn, the dorsal commissure, and lateral laminae V-VII in the region of the sacral parasympathetic nucleus. These experiments indicate that (1) electrical stimulation of abdominal nerves following surgical exposure is a useful method for tracing visceral afferent pathways and (2) afferent information from the pelvic viscera is received by neurons in specific areas of the dorsal horn.     

Dichotomizing unmyelinated afferents supplying pelvic viscera and perineum are rare in the sacral segments of the cat

We have tested the hypothesis that referred pain of pelvic viscera is elicited by dichotomizing branches of unmyelinated primary afferents projecting via the pelvic nerve to the viscera and through the pudendal nerve to the perineum where pelvic pain is commonly referred to. Using neurophysiological techniques 588 unmyelinated single units projecting into either nerve were recorded in the ventral (n = 228) and dorsal (n = 360) root S2. In each sample only one neurone sent an axon into both nerves. Thus, dichotomizing afferents account for less than 0.5% of the afferent neurones and appear to be an unlikely explanation for referred pain in this body area.     

躯体和内脏SP能初级传入神经元的定位研究

本文在后根节和脊髓两个水平对躯体(胫神经)和内脏(膀胱)初级传入系统进行同时定性、定位的研究。对后根节选用两种较敏感的双标技术(HRP结合PAP,Biotin-WGA结合免疫荧光)。结果表明,在后根节中被标记的胫神经和膀胱的初级传入神经元均有一部分同时显示SP样阳性反应。另外在分别切断盆内脏神经和胫神经的两组动物模型上进行脊髓的SP样免疫反应,通过两侧对比,观察到术侧灰质在相当于被切断神经的传入经路及终末区出现SP样反应的“脱落”现象,从而反证出躯体和内脏初级传入纤维中的SP能成分在中枢内的定位分布。本文首次提供了内脏初级传入在中枢内定位与定性相结合的研究结果。