Shigematsu K, Kamo H, Akiguchi I, Kimura H
Department of Neurology, Faculty of Medicine, Kyoto University, Japan.
J Neurosci Res. 1989 Nov;24(3):384-90. doi: 10.1002/jnr.490240307.
Successfully transplanted neurons and their sprouting processes were demonstrated by Phaseolus vulgaris leucoagglutinin (PHA) marking. PHA is transported anterogradely and readily reveals the post-transplantation growth of the neuronal processes. Suspensions of fetal central nervous tissue, prepared by dissociation of embryonic rat brain, were marked with PHA and then transplanted into the striatum of nonimmunosuppressed young adult rats. At various intervals thereafter (1 day, 2 days, 1 week, 2 weeks, 1 month, and 2 months), the animals were sacrificed for histological examination with PHA and tyrosine hydroxylase (TH) immunohistochemistry. Up to 2 weeks after transplantation, PHA immunohistochemistry was capable of demonstrating grafted neurons and their presumably regenerated neuronal processes. However, at periods 1 month or longer after transplantation, PHA immunohistochemistry was unreliable. Thus, the PHA marking method has limitations in terms of its retention period, when applied to the intracerebral transplantation of dissociated cell suspensions. Nevertheless, the method presented here can be utilized to study neuronal regeneration as well as the relationship between transplanted neurons and the host tissue.
通过菜豆白细胞凝集素(PHA)标记证明了神经元移植成功及其发芽过程。PHA可进行顺行运输,并能轻易显示移植后神经元突起的生长情况。将通过解离胚胎大鼠脑制备的胎儿中枢神经组织悬液用PHA标记,然后移植到未免疫抑制的成年幼鼠纹状体中。此后在不同时间间隔(1天、2天、1周、2周、1个月和2个月),处死动物进行PHA和酪氨酸羟化酶(TH)免疫组织化学的组织学检查。移植后长达2周,PHA免疫组织化学能够显示移植的神经元及其可能再生的神经元突起。然而,在移植后1个月或更长时间,PHA免疫组织化学结果不可靠。因此,当应用于解离细胞悬液的脑内移植时,PHA标记方法在保留期方面存在局限性。尽管如此,本文介绍的方法可用于研究神经元再生以及移植神经元与宿主组织之间的关系。