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菜豆白细胞凝集素(PHA-L):一种用于猫背外侧膝状核突触回路电子显微镜分析的神经解剖学示踪剂。

Phaseolus vulgaris leucoagglutinin (PHA-L): a neuroanatomical tracer for electron microscopic analysis of synaptic circuitry in the cat's dorsal lateral geniculate nucleus.

作者信息

Cucchiaro J B, Uhlrich D J

机构信息

Department of Neurobiology and Behavior, State University of New York, Stony Brook 11794.

出版信息

J Electron Microsc Tech. 1990 Aug;15(4):352-68. doi: 10.1002/jemt.1060150405.

Abstract

Phaseolus vulgaris leucoagglutinin (PHA-L) is a plant lectin that is anterogradely transported by neurons in the central nervous system. PHA-L is selectively taken up by cells at iontophoretic injection sites and, when immunohistochemically demonstrated, labels individual neurons completely, including their dendrites, axons, and terminal boutons. PHA-L is generally not taken up by fibers passing through the injection site and, because it produces a Golgi-like staining of even very fine axons over long distances, it is sometimes possible to light microscopically reconstruct individual neurons and their entire axon terminal arbors. When prepared for electron microscopy, the PHA-L-labeled terminals are densely and completely stained, allowing their synaptic relationships to be defined. These properties make PHA-L advantageous for studying the patterns of projection and the modes of termination of select groups of neurons in their target nuclei. We used PHA-L to study the extraretinal innervation of the cat's dorsal lateral geniculate nucleus, a thalamic visual center. Although much is known about the retinal contribution to geniculate synaptic circuitry, relatively little is known about other sources of innervation, even though these provide the majority of synaptic terminals in the nucleus (Guillery: Z. Zellforsch., 96:1-38, 39-48, 1969; Wilson et al.: Proc. R. Soc. Lond. [Biol.], 221:441-436, 1984). We used both light and electron microscopy to describe synaptic circuitry from three extraretinal sources of projections to the lateral geniculate nucleus: the visual cortex, the perigeniculate nucleus, and the parabrachial region of the brainstem. Cortical terminals labeled with PHA-L were small and formed asymmetrical synaptic contacts onto small-caliber dendrites of geniculate neurons. Perigeniculate terminals formed symmetrical synaptic contacts primarily onto small-caliber dendrites, but some synapses were also formed onto the proximal, retinorecipient portions of geniculate dendrites. Parabrachial terminals synaptically contacted the retinorecipient portions of dendritic appendages and shafts, small-caliber dendrites, and the specialized dendritic (F2) terminals of geniculate interneurons. The symmetry of the parabrachial synaptic contacts was variable and was related to the postsynaptic target. Contacts onto dendritic appendages were asymmetrical while those onto dendritic shafts and F2 terminals were symmetrical. Our data suggest that in unlabeled material these brainstem terminals would be difficult to distinguish from cortical or perigeniculate profiles. The positioning of the parabrachial input onto the retinorecipient portions of geniculate dendrites indicates that this projection is well situated to control primary retinal transmission through the nucleus, while the location of most cortical and perigeniculate innervations implicates them in secondary feedback interactions or other aspects of geniculate function.

摘要

菜豆白细胞凝集素(PHA-L)是一种植物凝集素,可被中枢神经系统中的神经元顺行运输。PHA-L在离子电渗注射部位被细胞选择性摄取,免疫组织化学显示时,它能完全标记单个神经元,包括其树突、轴突和终末小体。穿过注射部位的纤维通常不摄取PHA-L,并且由于它能对很长距离内甚至非常细的轴突产生类似高尔基染色的效果,有时可以通过光学显微镜重建单个神经元及其整个轴突终末分支。当制备用于电子显微镜观察时,PHA-L标记的终末被密集且完全染色,从而能够确定它们的突触关系。这些特性使PHA-L在研究特定神经元群在其靶核中的投射模式和终末方式方面具有优势。我们使用PHA-L研究猫背外侧膝状核(一个丘脑视觉中枢)的视网膜外神经支配。尽管对视网膜对膝状突触回路的贡献了解很多,但对其他神经支配来源了解相对较少,尽管这些来源提供了该核中大部分的突触终末(吉勒里:《细胞研究杂志》,96:1 - 38,39 - 48,1969;威尔逊等人:《伦敦皇家学会学报》[生物学],221:441 - 436,1984)。我们使用光学显微镜和电子显微镜来描述从三个视网膜外投射源到外侧膝状核的突触回路:视觉皮层、膝状周核和脑干臂旁区域。用PHA-L标记的皮层终末较小,在膝状神经元的小口径树突上形成不对称突触联系。膝状周终末主要在小口径树突上形成对称突触联系,但也有一些突触形成在膝状树突的近端视网膜接受部分上。臂旁终末与树突附属物和树突干的视网膜接受部分、小口径树突以及膝状中间神经元的特化树突(F2)终末形成突触联系。臂旁突触联系的对称性是可变的,并且与突触后靶点有关。与树突附属物的联系是不对称的,而与树突干和F2终末的联系是对称的。我们的数据表明,在未标记的材料中,这些脑干终末很难与皮层或膝状周轮廓区分开来。臂旁输入在膝状树突的视网膜接受部分上的定位表明,这种投射处于很好的位置来控制通过该核的初级视网膜传递,而大多数皮层和膝状周神经支配的位置表明它们参与二级反馈相互作用或膝状功能的其他方面。

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