Huang F L, Yoshida Y, Nakabayashi H, Young W S, Huang K P
Section on Metabolic Regulation, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
J Neurosci. 1988 Dec;8(12):4734-44. doi: 10.1523/JNEUROSCI.08-12-04734.1988.
Recently, we isolated 3 protein kinase C (PKC) isozymes from rat brain (Huang et al., 1986a). Using isozyme-specific antibodies for immunoblot, we have determined the relative levels of each isozyme in various regions of the rat brain (Huang et al., 1987b). The present paper describes the cellular distributions of PKC isozymes in rat brain as determined by light microscopic immunocytochemistry. Staining with PKC antibodies revealed strong immunoreactivities in neuronal somata and their dendrites and weak to no reaction in axon and the astroglial structures. In the cerebellum, the type I PKC antibodies stained the Purkinje cell bodies and dendrites; the type II PKC antibodies stained the granule cells; and the type III PKC antibody stained both Purkinje and granule cells. In the cerebral cortex, all antibodies stained neurons resembling pyramidal cells and their apical dendrites in layers II to VI, while layer I was nearly devoid of staining. However, the various isozyme-specific antibodies revealed distinct laminar distribution patterns of the positively stained neurons, and the type III PKC-positive neurons exhibited a higher density than those of type I or II PKC-positive ones, especially in layer II of cingulate (retrosplenial) and piriform cortices. In the hippocampal formation, both pyramidal cells of the hippocampus and granule cells of the dentate gyrus were stained by all PKC antibodies. Subcellularly, type III PKC appeared mostly in the cytoplasm of these neurons, whereas type I and II PKC seemed to associate with the nucleus as well. In the olfactory bulb, both type II and III PKC antibodies stained the periglomerular and granular cells, and the latter also stained the mitral cells. The distinct cellular and subcellular distribution of PKC isozymes suggests that each isozyme plays a unique role in the various neural functions.
最近,我们从大鼠脑中分离出了3种蛋白激酶C(PKC)同工酶(Huang等人,1986a)。使用同工酶特异性抗体进行免疫印迹,我们已经确定了大鼠脑各个区域中每种同工酶的相对水平(Huang等人,1987b)。本文描述了通过光学显微镜免疫细胞化学确定的大鼠脑中PKC同工酶的细胞分布。用PKC抗体染色显示,神经元胞体及其树突中有很强的免疫反应性,而轴突和星形胶质结构中反应较弱或无反应。在小脑中,I型PKC抗体染色浦肯野细胞体和树突;II型PKC抗体染色颗粒细胞;III型PKC抗体染色浦肯野细胞和颗粒细胞。在大脑皮层中,所有抗体都染色了II至VI层中类似锥体细胞及其顶端树突的神经元,而I层几乎没有染色。然而,各种同工酶特异性抗体显示出阳性染色神经元的不同分层分布模式,III型PKC阳性神经元的密度高于I型或II型PKC阳性神经元,尤其是在扣带回( retrosplenial)和梨状皮层的II层。在海马结构中,海马的锥体细胞和齿状回的颗粒细胞都被所有PKC抗体染色。在亚细胞水平上,III型PKC主要出现在这些神经元的细胞质中,而I型和II型PKC似乎也与细胞核有关。在嗅球中,II型和III型PKC抗体都染色了球周细胞和颗粒细胞,后者也染色了二尖瓣细胞。PKC同工酶独特的细胞和亚细胞分布表明,每种同工酶在各种神经功能中都发挥着独特的作用。