Ultrastructural localization of cAMP-dependent protein kinase subunits in regenerating rat hepatocytes using immunogold electron microscopy.

S P Squinto, D C Kelley-Geraghty, M R Kuettel, R A Jungmann
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Abstract

We have used the indirect colloidal immunogold technique to examine the ultrastructural localization and distribution of the regulatory subunits (RI and RII) and catalytic subunit (C) of cAMP-dependent protein kinase (cA-PK) in hepatocyte nuclei during the prereplicative phase of rat liver regeneration. The technique allowed the localization of all three subunits of cA-PK in hepatocyte nuclei. Morphometric quantitation of the relative staining intensity of nuclear antigen indicated a marked increase of immunogold staining of nuclear RI and C 16 hours following partial hepatectomy. In contrast, staining of nuclear RII was not affected. These immunocytochemical observations correlate well with our previous data (Laks, M.S. et al. (1981) J. Biol. Chem. 256, 8775-8785) obtained by biochemical methods indicating a modulation of cA-PK activity after partial hepatectomy.

免疫金电镜观察再生大鼠肝细胞camp依赖性蛋白激酶亚基的超微结构定位。
我们利用间接胶体免疫金技术研究了大鼠肝再生增殖前期肝细胞核中camp依赖性蛋白激酶(cA-PK)的调控亚基(RI和RII)和催化亚基(C)的超微结构定位和分布。该技术允许在肝细胞核中定位cA-PK的所有三个亚基。核抗原相对染色强度的形态学定量显示,核RI和核C在部分肝切除术后16小时免疫金染色明显增加。相反,核RII染色不受影响。这些免疫细胞化学观察结果与我们以前的数据相吻合(Laks, M.S. et al.(1981))。化学,256,8775-8785)通过生化方法获得,表明部分肝切除术后cA-PK活性的调节。
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