Dingemanse M A, de Boer P A, Moorman A F, Charles R, Lamers W H
Department of Anatomy and Embryology, University of Amsterdam, The Netherlands.
Differentiation. 1994 May;56(3):153-62. doi: 10.1046/j.1432-0436.1994.5630153.x.
The onset of transcription and mRNA accumulation of two liver-specific genes, carbamoylphosphate synthase (CPS) and phosphoenolpyruvate carboxykinase (PEPCK) in individual embryonic rat hepatocytes was investigated with in situ hybridization. In vitro CPS and PEPCK mRNAs can be induced prematurely in monolayer cultures of embryonic rat hepatocytes by glucocorticosteroids and cyclic AMP, i.e. the hormones that also regulate the expression of these genes in vivo. Upon exposure to hormones the cultures showed an interhepatocyte heterogeneity in CPS and PEPCK mRNA content. The pattern of accumulation of nuclear CPS mRNA-precursors indicates that this heterogeneity is generated by intercellular differences in the timing of the onset of transcription. However, under induced steady-state conditions the heterogeneity in the hepatocyte population persisted. The degree of heterogeneity is inversely related to the half life of the gene product (i.e. higher for PEPCK than for CPS and higher for mRNAs than for the respective proteins) and to the concentrations of inducing hormones. Accordingly, the interhepatocyte heterogeneity was most pronounced for the nuclear CPS mRNA-precursor. In contrast, no intercellular differences in the rate of degradation of the mRNAs were seen. These observations reveal that although all hepatocytes can and do express the genes, transcription of a gene in a particular cell is a discontinuous process.
采用原位杂交技术研究了个体胚胎大鼠肝细胞中两个肝脏特异性基因——氨甲酰磷酸合成酶(CPS)和磷酸烯醇式丙酮酸羧激酶(PEPCK)的转录起始及mRNA积累情况。在体外,糖皮质激素和环磷酸腺苷(即体内也调节这些基因表达的激素)可使胚胎大鼠肝细胞单层培养物中的CPS和PEPCK mRNA过早诱导产生。暴露于激素后,培养物中肝细胞间的CPS和PEPCK mRNA含量存在异质性。核CPS mRNA前体的积累模式表明,这种异质性是由转录起始时间的细胞间差异产生的。然而,在诱导的稳态条件下,肝细胞群体中的异质性仍然存在。异质性程度与基因产物的半衰期呈负相关(即PEPCK的半衰期比CPS长,mRNA的半衰期比相应蛋白质长),且与诱导激素的浓度有关。因此,核CPS mRNA前体的肝细胞间异质性最为明显。相比之下,未观察到mRNA降解速率的细胞间差异。这些观察结果表明,尽管所有肝细胞都能够且确实表达这些基因,但特定细胞中基因的转录是一个不连续的过程。