Lamas E, Kahn A, Guillouzo A
J Histochem Cytochem. 1987 May;35(5):559-63. doi: 10.1177/35.5.3104450.
In situ hybridization on tissue sections was used to detect mRNAs present at low concentrations during metabolic adaptation and azo dye carcinogenesis in rat liver. The method consisted of hybridizing the slices at relatively high stringency with [35S]-labeled single-stranded probes derived from cDNA insert clones into the M13 phage. L-pyruvate kinase mRNA was proved to be present at very low concentrations in hepatocytes of fasted rats and to be relatively abundant in all hepatocytes after 18 hr of refeeding on a carbohydrate-rich diet. Aldolase A mRNA concentrations have been previously shown to increase markedly in liver of 3'-methyl DAB-fed rats, with a maximum at the fourth week. We demonstrate here, using our in situ hybridization technique, that this phenomenon is not due to re-expression of this "fetal marker" in hepatocytes but to its abundancy in proliferating small cells (i.e., so-called oval and transitional cells). Small amounts were also detected in sinusoidal cells. In normal liver, aldolase A mRNAs were detected only in some sinusoidal cells.
采用组织切片原位杂交技术，检测大鼠肝脏在代谢适应和偶氮染料致癌过程中低浓度存在的mRNA。该方法包括将切片与来源于插入M13噬菌体的cDNA克隆的[35S]标记单链探针在相对高严谨度下进行杂交。结果表明，禁食大鼠肝细胞中L-丙酮酸激酶mRNA浓度极低，而在富含碳水化合物的食物再喂养18小时后，所有肝细胞中该mRNA相对丰富。先前已表明，喂食3'-甲基二氨基联苯（3'-methyl DAB）的大鼠肝脏中醛缩酶A mRNA浓度显著增加，在第四周达到最高。我们在此利用原位杂交技术证明，这种现象并非由于该“胎儿标志物”在肝细胞中重新表达，而是由于其在增殖的小细胞（即所谓的卵圆细胞和过渡细胞）中含量丰富。在肝血窦细胞中也检测到少量该mRNA。在正常肝脏中，仅在一些肝血窦细胞中检测到醛缩酶A mRNA。
