Shimohama S, Saitoh T, Gage F H
Department of Neurosciences, School of Medicine (0624), University of California, San Diego, La Jolla 92093-0624.
Hippocampus. 1993 Jan;3(1):43-55. doi: 10.1002/hipo.450030105.
The cellular and regional distribution of the four protein kinase C (PKC) isoforms in the rat hippocampal formation and the response of PKC to lesions were determined by employing immunohistochemical and immunochemical techniques with antibodies specific to PKC(alpha), -(beta I), -(beta II), and -(gamma). PKC(alpha) intensely stained the periphery of the pyramidal cell in the stratum pyramidale. The granule cells, glial cells, and mossy fibers were anti-PKC(alpha) negative. The cytoplasm, axons, and dendrites of basket cells and interneurons in the hilus were labeled with anti-PKC(alpha). Anti-PKC(beta I) immunoreactivity was localized on the periphery of pyramidal cells and interneurons of the hilus, as well as the oriens, radiatum, and molecular layers of the CA regions. Anti-PKC(beta II) immunoreactivity was mainly cytoplasmic, extending into the dendrites in the hippocampal pyramidal cells and the dentate granule cells, and also in some glial cells. In the stratum radiatum of the CA1, anti-PKC(gamma) immunoreactivity localized to the pyramidal cell cytoplasm, extending into the dendrites. Following fimbria-fornix (FF) lesions, the anti-PKC(alpha) and -(beta I) staining of the pyramidal cell periphery was markedly reduced. The anti-PKC(gamma) staining of the pyramidal and granular cells of the dentate gyrus was reduced whereas the interneuron staining in the hilus was increased. In the FF-lesioned hippocampus, anti-PKC(alpha) and anti-PKC(beta II) labeled reactive glial cells, whereas anti-PKC(beta I) and -(gamma) did not. Quantitative Western blot analysis revealed a dramatic increase in the particulate/total PKC for all isozyme forms, although the total levels of PKC, except PKC(gamma), did not change following FF lesions. The PKC(gamma) concentration doubled after FF lesions. Perforant path lesions resulted in a marked alteration in the neuronal staining in dentate gyrus with anti-PKC(alpha) and -(beta I) and in increased numbers of anti-PKC(alpha)- and anti-PKC(beta II)-positive glial cells. Anti-PKC(gamma) staining did not change noticeably. The total PKC concentration did not change for isozymes alpha, beta I, and gamma, but PKC (beta II) concentration increased by 48% following perforant path lesions as detected by Western blot analysis. The particulate/total PKC decreased for all four isozymes although the reduction in PKC(beta I) concentration was not statistically significant. This change in PKC compartmentalization is in marked contrast to an increased level of particulate PKC following FF lesions. Thus, the effects of deafferentation and deafferentation for each PKC isoform were different.
采用针对蛋白激酶C（PKC）α、βI、βII和γ亚型的特异性抗体，运用免疫组织化学和免疫化学技术，确定了大鼠海马结构中四种PKC亚型的细胞和区域分布以及PKC对损伤的反应。PKCα强烈染色锥体层中锥体细胞的周边。颗粒细胞、胶质细胞和苔藓纤维抗PKCα呈阴性。门区篮状细胞和中间神经元的细胞质、轴突和树突被抗PKCα标记。抗PKCβI免疫反应定位于门区锥体细胞和中间神经元的周边，以及CA区的梭形层、辐射层和分子层。抗PKCβII免疫反应主要位于细胞质，延伸至海马锥体细胞和齿状颗粒细胞的树突，以及一些胶质细胞。在CA1的辐射层，抗PKCγ免疫反应定位于锥体细胞细胞质，延伸至树突。海马伞-穹窿（FF）损伤后，锥体细胞周边的抗PKCα和βI染色明显减少。齿状回锥体细胞和颗粒细胞的抗PKCγ染色减少，而门区中间神经元染色增加。在FF损伤的海马中，抗PKCα和抗PKCβII标记反应性胶质细胞，但抗PKCβI和γ未标记。定量蛋白质免疫印迹分析显示，所有同工酶形式的颗粒状/总PKC显著增加，尽管除PKCγ外，PKC的总水平在FF损伤后未发生变化。FF损伤后PKCγ浓度翻倍。穿通通路损伤导致齿状回中抗PKCα和βI的神经元染色明显改变，以及抗PKCα和抗PKCβII阳性胶质细胞数量增加。抗PKCγ染色无明显变化。同工酶α、βI和γ的总PKC浓度未改变，但蛋白质免疫印迹分析检测显示，穿通通路损伤后PKCβII浓度增加48%。所有四种同工酶的颗粒状/总PKC均降低，尽管PKCβI浓度的降低无统计学意义。这种PKC区室化的变化与FF损伤后颗粒状PKC水平的增加形成显著对比。因此，去传入和去传入对每种PKC亚型的影响是不同的。
