大鼠前庭核向脑干呕吐区的间接投射

前庭信息调节内脏活动的神经通路是产生运动病的结构基础,但至今尚未明了。本研究旨在探讨接受初级前庭传入信息的前庭核到脑干呕吐中枢的神经通路。向大鼠前庭内侧核和前庭下核内注入顺行追踪剂生物素化葡聚糖胺(BDA),向脑干呕吐区注入逆行追踪剂荧光金(FG),用荧光组织化学方法显示BDA顺行标记纤维和终末,在荧光显微镜下观察BDA顺行标记纤维和终末与荧光金逆行标记细胞的重叠区域。结果发现在延髓背侧巨细胞旁核(DPGi)、巨细胞网状核(Gi)和小细胞网状核(PCRt)有顺行标记纤维与逆行标记细胞的重叠。表明前庭核可能经DPGi、Gi和PCRt向呕吐区有间接投射,此结果为进一步揭示前庭信号引发恶心、呕吐的神经机制提供了形态学依据。     

大鼠前庭脊核和X细胞群向脑桥核的直接投射

目的:应用菜豆白细胞凝集素(PHA-L)顺行追踪和荧光金(FG)逆行追踪技术研究前庭脊核和X细胞群向脑桥核的直接投射.方法:SD大鼠随机分为PHA-L注射组和FG注射组.将顺行神经追踪剂PHA-L电泳至前庭脊核和X细胞群,逆行神经追踪剂FG分别电泳至脑桥核的外侧亚核和内侧亚核,动物存活7 d,灌流固定后,脑干作冠状冷冻切片,然后进行免疫组织化学显色.结果:PHA-L注射于前庭脊核后,顺行标记纤维和终末主要分布在对侧脑桥核的外侧亚核、内侧亚核及脑桥网状被盖核;FG分别注射于脑桥核的外侧和内侧亚核后,逆行标记细胞仅分布在对侧前庭脊核和X细胞群.结论:前庭脊核和X细胞群向对侧脑桥核的外侧和内侧亚核有直接的纤维投射,该投射可能与前庭-眼反射的调节有关.     

前庭神经核纤维与投射至纹状体的束旁核神经元的突触联系

目的：观察发自前庭神经内侧核的纤维末梢与投射至纹状体的丘脑束旁核神经元的突触联系。方法：采用15只Wistar大鼠，应用顺行和逆行标记技术，免疫组织化学和免疫电镜方法。结果：将CTb单侧注入纹状体，同时将BDA注入同侧的前庭神经内侧核。在束旁核发现了CTb标记神经元和BDA标记轴突终末，BDA标记纤维和终末存在于外侧束旁核整个长度的背侧2／3区，而CTb标记神经元也存在于外侧束旁核背侧2／3区，2种标记相互重叠。电镜下可见标记终末与标记神经元形成非对称性的轴-体和轴-树突触。结论：由前庭神经内侧核发出的投射纤维在束旁核与投射至纹状体的束旁核神经元之间存在着非对称性的突触联系。     

大鼠杏仁复合体向听皮层的神经投射

目的：研究大鼠杏仁复合体一听皮层的纤维投射。方法：HRP神经追踪方法结合微电泳技术，以及核黄逆行荧光标记技术。结果：HRP注射到听皮层，同侧杏仁外侧核、杏仁基底核、杏仁前区和杏仁前皮层观察到逆行标记细胞；HRP注射到杏仁外侧核和杏仁基底核，在同侧听皮层出现顺行标记纤维；核黄注入到听皮层，在同侧杏仁外侧核和杏仁基底核发现逆行标记细胞。结论：大鼠听皮层接受同侧杏仁复合体的神经投射。     

大鼠外侧楔核向小脑投射的追踪研究——PHA-L法

用菜豆白细胞凝集素(PHA-L)顺行追踪大鼠外侧楔核向小脑的投射。将PHA-L溶液电泳导入大鼠的外侧楔核后,用ABC免疫组织化学法可以清楚地显示出被标记神经细胞、纤维及终末的形态。结果表明,大鼠外侧楔核的投射纤维经同侧绳状体进入小脑。终末区主要包括小脑前、后叶的双侧蚓部、同侧前叶的半球部、同侧Ⅷ小叶的旁蚓部、Ⅸa小叶及同侧旁正中小叶的内侧部。Ⅳ-Ⅴ小叶上终末密度最高,并形成三条平行于正中矢状平面的终末带。在同侧脚Ⅰ及Ⅸb小叶上发现少量终末。结果还提示,外侧楔核无纤维终末至小脑中央核。     

黄雀延髓层状核的传出纤维联系──HRP法研究

向４２只黄雀的延髓展状核加压注入或微电泳泳入ＨＲＰ后，在同侧的脑桥上橄榄核、对侧的脑桥外侧丘系腹核及中脑外侧核背侧部内见到密集的顺行标记终末或纤维；在双侧的延髓巨细胞核出现了大量的逆行标记细胞；在对侧的层状核内既有密集的标记终末又有一些标记细胞．结果表明：层状核的传出纤维投射到同侧上橄榄核，经外侧丘系走行至对侧的外侧丘系腹核和中脑外侧核背侧部，双侧的层状核之间也有交互投射。此外，层状核接受耳蜗亚核──双侧巨细胞核的传入投射．因此，层状核是听觉上行通路中的第二级中继站．     

黄喉鹀耳蜗核的传出投射

用HRP顺行追踪方法,研究黄喉鵐(emberiza elegans)的两对耳蜗核,即角状核和巨细胞核的传出投射.将HRP注入角状核,在双侧上橄榄核,对侧外侧丘系核腹侧部,外侧丘系腹核及中脑背外侧核的背侧1/4的区域见到顺行标记纤维或终末.将HRP注入巨细胞核,标记纤维或终末分布于双侧层状核;标记细胞分布于同侧上橄榄核.结果表明:角状核投射至双侧上橄榄核,对侧外侧丘系核腹侧部,外侧丘系腹核及中脑背外侧核的背侧部.巨细胞核投射至双侧层状核.此外,巨细胞核接受同侧上橄榄核的传人,它可能是一条听觉的反馈回路.     

下丘脑室旁核后叶加压素神经元向迷走神经复合体和脊髓的投射——HRP 逆行追踪和免疫组化结合法研究

本文报道用 HRP 逆行追踪和 PAP 免疫组织化学结合法研究了大鼠室旁核向迷走神经复合体和脊髓的投射及其化学性质。脊髓一侧注射 HRP 后,室旁核 HRP 标记细胞集中于小细胞尾侧区,以其内侧部,外侧部和背部最多。在脊髓各组中比较,胸段组出现的 HRP 标记细胞最多,腰段组次之,骶段组最少。迷走神经复合体注射 HRP 后,HRP 标记细胞的分布与脊髓注射后的分布相似,但标记细胞较稀疏。结果表明,室旁核(主要是小细胞尾侧区)发出下行纤维投射至迷走神经复合体和脊髓。室旁核 HRP 标记细胞中,有一些细胞呈后叶加压素免疫反应阳性。脊髓组双标细胞多位于 HRP 标记细胞的主要分布区,即内侧部、外侧部和背部。迷走神经复合体组双标细胞的分布与脊髓组的分布基本相同,仅双标细胞较稀少,约占脊髓组 HRP 标记细胞的19.2%,占迷走神经复合体组 HRP 标记细胞的20.3%。从而为室旁核后叶加压素神经元投射到迷走神经复合体和脊髓提供了直接证据。     

猫红核和中脑中央灰质向下橄榄复合体的投射——WGA-HRP顺行追踪研究

分别向红核和中脑中央灰质注入5%WGA-HRP,顺行追踪它们向下橄榄复合体(IOC)的投射。本文证明:1.两者都向同侧IOC投射,且在IOC内未见逆行标记细胞。2.红核投射至下橄榄主核的背侧板及内侧副橄榄核尾侧半的背端部;中央灰质则投射至内侧副橄榄核的全部、主核腹侧板及腹侧板与背侧板的腹端移行部。其中,中央灰质向内侧副橄榄核的投射及红核向主核背侧板的投射,标记终末异常密集,其他部分则较稀疏。3.本文所见标记区,与文献报道脊髓向IOC的投射区有一定重叠。与以往报道的红核及中央灰质向IOC的投射区相比,范围较大。     

大鼠外侧巨细胞旁网状核的传出纤维联系——HRP法研究

用HRP轴■顺、逆行追踪法观察了大鼠外侧巨细胞旁网状核(PGCL)的传出纤维联系。结果表明:①PGCL经轴■顺行传递,可投射到与痛觉及其调控有关的核团,如脊髓背角、三叉神经脊束核、导水管周围灰质、束旁核、外侧颈核、脑干网状结构核群等;PGCL还投射到与调节内脏活动有关的孤束核、迷走神经背核、导水管周围灰质背份、臂旁核、脊髓侧角等;也向三叉神经运动核、下丘等处发出投射纤维。②腰髓注射HRP后,在PGCL中见有较多的标记细胞,主要分布在锥体束外侧和面神经核腹内侧区域,部分细胞亦见于锥体束内及PGCL靠近软脑膜处。③向下丘和孤束核注入HRP以作往返印证,在PGCL见到标记细胞和纤维。     

Direct projection from the dorsal hypothalamic area to the nucleus raphe pallidus: a study using anterograde transport with Phaseolus vulgaris leucoagglutinin in the rat

Summary A hypothalamic projection to the nucleus raphe pallidus of the medulla was examined using the anterograde tracing technique based on Phaseolus vulgaris leucoagglutinin (PHA-L) in the rat. After the iontophoretic application of PHA-L to the dorsal hypothalamic area, labeled fibers that finally ended in the nucleus raphe pallidus were observed descending through the most medial part of the ventral tegmental area and the nucleus reticularis tegmenti pontis to reach the medial aspect of the pyramid. Many varicose fibers forming a loose plexus were observed in the nucleus raphe pallidus, especially ventrally. The ventral surface of the pyramid and the most ventral region of the nucleus reticularis paragigantocellularis lateralis (PGCL) contained labeled varicose fibers. At the electron microscopic level, the labeled profiles in the nucleus raphe pallidus were small-sized unmyelinated axons and axon terminals. Labeled axon terminals containing spherical synaptic vesicles formed synapses on spine-like protrusions or small-sized dendritic shafts. These results strongly indicate that neurons in the dorsal hypothalamic area have a direct connection with neurons in the nucleus raphe pallidus and the ventral part of the PGCL. The possible involvement of this pathway in cardiovascular regulation was discussed.     

小鼠脑干交感运动前区前阿黑皮素能神经传入投射的实验研究

目的:研究脑干交感运动前区的前阿黑皮素能神经纤维的分布及其传入投射。方法:以免疫组化法检测前阿黑皮素能神经纤维在脑干交感运动前区的分布;将霍乱毒素B微量注入小鼠脑干交感运动前区进行逆行追踪。结果:α-MSH(α-melanocyte-stimulating hormone)能神经终末密集分布于交感运动前区内,同时则未见AgRP(agouti-related protein)能神经终末,显示交感运动前区仅接受前阿黑皮素(POMC)能神经的单向调节。逆行追踪实验显示脑干交感运动前区的POMC能纤维投射来自于下丘脑弓状核/视交叉后区,而非孤束核尾侧部;统计结果表明下丘脑弓状核/视交叉后区向脑干交感运动前区的神经投射中有近一半为POMC能投射。结论:小鼠下丘脑的POMC能神经元可直接投射到脑干交感运动前区,该神经通路在能量平衡的调节中可能发挥重要的作用。     

亮氨酸脑啡肽样和P物质样免疫反应物在大鼠延髓迷走神经背核簇和网状结构内的分布

用PAP法研究亮氨酸脑啡肽样(L—ENK—LI)和P物质样(SP—LI)免疫反应物在大鼠延髓迷走神经背核簇和网状结构内的分布。证明在孤束核、迷走神经背核、外侧网状核外侧部及其外侧的区域内有大量的L-ENK样终末和纤维,在背侧和腹侧网状核之间的移行区内有中等量的分布,其余区内为少量。L-ENK样胞体在孤束核、咀侧腹外侧网状核、巨细胞网状核的腹侧部和α部、外侧旁巨细胞核以及中缝大核内均有许多分布,在迷走神经背核的尾侧部、背侧和腹侧网状核之间的移行区有中等量,其余区内为少量。SP样反应物的分布与L-ENK样物类似,但其终末和纤维的数量较L-ENK者略少,阳性胞体的数量除了在中缝核及外侧旁巨细胞核内侧端中的量较L-ENK样胞体多以外,在其余区内均较少。     

Arcuate nucleus projections to brainstem regions which modulate nociception

Anterograde tracing studies were conducted in order to identify efferents from the arcuate nucleus, which contains the hypothalamic opiocortin neuronal pool. Phaseolus vulgaris leucoagglutinin (PHA-L) was stereotaxically iontophoresed into the arcuate nucleus and the terminal fields emanating from the labelled perikarya were identified immunocytochemically. PHA-L-immunoreactive (-ir) fibers were identified in nucleus accumbens, lateral septal nucleus, bed nucleus of the stria terminalis, medial and lateral preoptic areas, anterior hypothalamus, amygdaloid complex, lateral hypothalamus, paraventricular nucleus, zona incerta, dorsal hypothalamus, periventricular gray, medial thalamus and medial habenula. In the brainstem, arcuate terminals were identified in the periaqueductal gray (PAG), dorsal raphe nucleus (DRN), nucleus raphe magnus (NRM), nucleus raphe pallidus, locus coeruleus, parabrachial nucleus, nucleus reticularis gigantocellularis pars alpha, nucleus tractus solitarius and dorsal motor nucleus of the vagus nerve. Dual immunostaining was used to identify the neurochemical content of neurons in arcuate terminal fields in the brainstem. Arcuate fiber terminals established putative contacts with serotonergic neurons in the ventrolateral PAG, DRN and NRM and with noradrenergic neurons in periventricular gray, PAG and locus coeruleus. In the PAG, arcuate fibers terminated in areas with neurons immunoreactive to substance P, neurotensin, enkephalin and cholecystokinin (CCK) and putative contacts were identified with CCK-ir cells. This study provides neuroanatomical evidence that putative opiocortin neurons in the arcuate nucleus influence a descending system which modulates nociception.     

Neurons in the rat medulla oblongata containing neuropeptide Y-, angiotensin II-, or galanin-like immunoreactivity project to the parabrachial nucleus.

Projections from the medulla to the parabrachial complex of the rat were examined for their content of neuropeptide Y-, angiotensin II- or galanin-like immunoreactivity using combined retrograde tracing and immunohistochemical techniques. Rhodamine-labelled latex microspheres were stereotaxically injected into discrete nuclei of the parabrachial complex. After survival of two to five days, colchicine (100 micrograms in 10 microliters saline) was injected into the cisterna magna. One day later, rats were perfused and the brainstems were prepared for visualization of the retrograde tracer and immunoreactivity of one of the three peptides. Retrograde labelling verified that the area postrema, nucleus of the tractus solitarius, caudal spinal nucleus of the trigeminal nerve, parvocellular reticular nucleus, and ventrolateral medulla including the rostral ventrolateral medulla and nucleus paragigantocellularis project to the lateral parabrachial and K?lliker-Fuse nuclei. While most projections were primarily ipsilateral, a small proportion of the projections from the ventrolateral medulla was bilateral. Neurons containing neuropeptide Y-like immunoreactivity were found in the caudal and intermediate nucleus of the tractus solitarius, dorsal to the lateral reticular nucleus and in the nucleus paragigantocellularis. After bilateral microsphere injections into the lateral parabrachial and K?lliker-Fuse nuclei, double-labelled neurons were found dorsal to the lateral reticular nucleus of caudal and intermediate medullary levels, at the ventral surface of the medulla at intermediate levels and in the nucleus paragigantocellularis at rostral levels. Neurons with angiotensin II-like immunoreactivity were observed at the dorsomedial border of the caudal and intermediate nucleus of the tractus solitarius, in the area postrema and in the lateral reticular nucleus and nucleus paragigantocellularis. Of these neurons, small numbers in the nucleus of the tractus solitarius and ventrolateral medulla also projected to the lateral parabrachial and K?lliker-Fuse nuclei. Neurons containing galanin-like immunoreactivity were found in the caudal nucleus of the tractus solitarius, the area postrema, the spinal trigeminal nucleus, the raphe nuclei (pallidus and obscurus), the nucleus paragigantocellularis and dorsal to the lateral reticular nucleus. Of these cells, double-labelled neurons were found in the commissural and medial subdivisions of the caudal nucleus of the tractus solitarius and in the rostral ventrolateral medulla including the ventral surface and the nucleus paragigantocellularis. The results suggest that neuropeptide Y, angiotensin II and galanin may serve as neurochemical messengers in pathways from the medulla to the parabrachial complex. The location of double-labelled neurons suggests that the information relayed by these neurons is related to autonomic activity.     

Tachykinin immunoreactivity in terminals of trigeminal afferent fibers in adult and fetal monkey thalamus

Summary Immunocytochemistry of fetal and adult monkey thalamus reveals a dense concentration of tachykinin immunoreactive fibers and terminals in the dorsolateral part of the VPM nucleus in which the contralateral side of the head, face and mouth is represented. The immunoreactive fibers enter the VPM nucleus from the thalamic fasciculus and electron microscopy reveals that they form large terminals resembling those of lemniscal axons and terminating in VPM on dendrites of relay neurons and on presynaptic dendrites of interneurons. Double labeling strategies involving immunostaining for tachykinins after retrograde labeling of brainstem neurons projecting to the VPM failed to reveal the origin of the fibers. The brainstem trigeminal nuclei, however, are regarded as the most likely sources of the VPM-projecting, tachykinin positive fibers.Abbreviations AB ambiguus nucleus - AN abducens nucleus - C cuneate nucleus - CD dorsal cochlear nucleus - CL central lateral nucleus - CM centre médian nucleus - D dendrite - DR dorsal raphe - DV dorsal vagal nucleus - EC external cuneate nucleus - FM medial longitudinal fasciculus - FN facial nucleus - G gracile nucleus - Gc gigantocellular reticular formation - HN hypoglossal nucleus - ICP inferior cerebellar peduncle - IO inferior olivary complex - LC locus coeruleus - LL lateral lemniscus - LM medial lemniscus - M5 motor trigeminal nucleus - NS solitary nucleus - OS superior olivary complex - P dendritic protrusion - Pb parabrachial nucleus - Pc parvocellular reticular formation - PLa anterior pulvinar nucleus - Pp prepositus hypoglossi nucleus - Ps presynaptic region - Py pyramidal tract - P5 principal sensory trigeminal nucleus - R reticular nucleus - RF reticular formation - RL lateral reticular nucleus - S5 spinal trigeminal nucleus - T terminal - T5 spinal trigeminal tract - VL lateral vestibular nucleus - VM medial vestibular nucleus - VMb basal ventral medial nucleus - VPI ventral posterior inferior nucleus - VPL ventral posterior lateral nucleus - VPM ventral posterior medial nucleus - VR ventral raphe - VS superior vestibular nucleus - VSp spinal vestibular nucleus - ZI zona incerta - 5 trigeminal nerve - 6 abducens nerve - 7 facial nerve     

猫内脏大神经一级传入纤维在脊髓灰质和薄束核中的分布

本文共用猫14只,取1～1.5mg HRP溶于7～10μl蒸馏水中,注入一侧的腹腔神经节或内脏大神经中,采用TMB成色法,观察跨神经节传递的一级内脏传人纤维在中枢神经系的分布。标记的一级内脏感觉纤维经后根进入脊髓后,绝大多数先行于背外侧束(或Lissauer束)中,少数进入后索上行。自背外侧束间断地发出内、外侧投射纤维,包绕着后角的内、外侧缘。外侧投射纤维在数量上比内侧的多,止于Ⅰ、Ⅴ、Ⅶ层和中央管周围。进入中间外侧核的纤维,再沿颅尾方向分开纵行,与交感节前细胞的纵向树突紧密平行排列。内侧投射纤维主要止于中央管周围区域。行于后索的纤维,止于闩平面以下薄束核的腹外侧部。     

Selective anterograde tracing of the individual serotonergic and nonserotonergic components of the dorsal raphe nucleus projection to the vestibular nuclei

It is well known that the dorsal raphe nucleus (DRN) sends serotonergic and nonserotonergic projections to target regions in the brain stem and forebrain, including the vestibular nuclei. Although retrograde tracing studies have reported consistently that there are differences in the relative innervation of different target regions by serotonergic and nonserotonergic DRN neurons, the relative termination patterns of these two projections have not been compared using anterograde tracing methods. The object of the present investigation was to trace anterogradely the individual serotonergic and nonserotonergic components of the projection from DRN to the vestibular nuclei in rats. To trace nonserotonergic DRN projections, animals were pretreated with the serotonergic neurotoxin 5,7-dihydroxytryptamine (5,7-DHT), and then, after 7 days, the anterograde tracer biotinylated dextran amine (BDA) was iontophoretically injected into the DRN. In animals treated with 5,7-DHT, nonserotonergic BDA-labeled fibers were found to descend exclusively within the ventricular plexus and to terminate predominantly within the periventricular aspect of the vestibular nuclei. Serotonergic DRN projections were traced by injecting 5,7-DHT directly into DRN, and amino-cupric-silver staining was used to visualize the resulting pattern of terminal degeneration. Eighteen hours after microinjection of 5,7-DHT into the DRN, fine-caliber degenerating serotonergic terminals were found within the region of the medial vestibular nucleus (MVN) that borders the fourth ventricle, and a mixture of fine- and heavier-caliber degenerating serotonergic terminals was located further laterally within the vestibular nuclear complex. These findings indicate that fine-caliber projections from serotonergic and nonserotonergic DRN neurons primarily innervate the periventricular regions of MVN, whereas heavier-caliber projections from serotonergic DRN neurons innervate terminal fields located in more lateral regions of the vestibular nuclei. Thus, serotonergic and nonserotonergic DRN axons target distinct but partially overlapping terminal fields within the vestibular nuclear complex, raising the possibility that these two DRN projection systems are organized in a manner that permits regionally-specialized regulation of processing within the vestibular nuclei.