Kim H M, Hirota S, Chung H T, Onoue H, Ito A, Morii E, Hirata T, Ohno S, Osada S, Kitamura Y
Department of Pathology, Osaka University Medical School, Japan.
J Mol Neurosci. 1993 Winter;4(4):245-53. doi: 10.1007/BF02821556.
In the central nervous system (CNS), the expression of protein kinase C (PKC) genes is strictly controlled by the developmental stage. We have examined the expression of PKC genes (cPKC alpha, beta, gamma, and nPKC delta, epsilon) in the process of the postnatal development in normal (+/+) C57BL/6 and microphthalmic (mi/mi) C57BL/6 mouse brains by Northern blotting and in situ hybridization. By Northern blotting, the expression level of cPKC gamma mRNA in mi/mi mice was significantly lower than that in +/+ littermates at d 9, 13, and 17. By in situ hybridization analysis, cPKC gamma mRNA-positive cells were detected in hippocampal and Purkinje cells in +/+ and mi/mi mice, but the magnitude of the signals in mi/mi mice was lower than that of +/+ mice, and the number of positive cells was smaller, whereas other isozymes (cPKC alpha, beta, and nPKC delta, epsilon) showed no significant difference between normal and mi/mi mice. The neuronal morphometric analysis by anti-P400 antibody revealed the same number and expression level of P400 protein in cerebellar Purkinje cells compared with +/+ mice. These results indicate that the deficiency of mi gene product causes the delayed expression of the cPKC gamma gene.
在中枢神经系统(CNS)中,蛋白激酶C(PKC)基因的表达受到发育阶段的严格调控。我们通过Northern印迹法和原位杂交技术,检测了正常(+/+)C57BL/6和小眼(mi/mi)C57BL/6小鼠脑在出生后发育过程中PKC基因(cPKCα、β、γ和nPKCδ、ε)的表达情况。通过Northern印迹法,在出生后第9天、13天和17天,mi/mi小鼠中cPKCγ mRNA的表达水平显著低于其+/+同窝小鼠。通过原位杂交分析,在+/+和mi/mi小鼠的海马和浦肯野细胞中均检测到cPKCγ mRNA阳性细胞,但mi/mi小鼠中的信号强度低于+/+小鼠,且阳性细胞数量较少,而其他同工酶(cPKCα、β和nPKCδ、ε)在正常小鼠和mi/mi小鼠之间无显著差异。用抗P400抗体进行的神经元形态计量分析显示,与+/+小鼠相比,小脑浦肯野细胞中P400蛋白的数量和表达水平相同。这些结果表明,mi基因产物的缺乏导致cPKCγ基因的表达延迟。