Szentirmay Z, Ishizaka Y, Ohgaki H, Tahira T, Nagao M, Esumi H
Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan.
Oncogene. 1990 May;5(5):701-5.
Expression of the ret proto-oncogene (proto-ret) in rat conceptus tissues during development was examined by in situ hybridization using photobiotin-labeled oligodeoxyribonucleic acid probes corresponding to regions coding for the kinase and transmembrane domains of proto-ret gene product. High levels of the proto-ret transcripts were detected in the cytotrophoblasts in the placenta in the mid-gestational period (days 10 and 11), but on day 14 of gestation, when the placenta was undergoing morphological changes, transcripts could no longer be detected in the trophoblasts. These results suggest that the increased expression of proto-ret may be associated with the proliferation and/or differentiation of trophoblast cells at a specific stage. Improvements in the in situ hybridization technique by introduction of an ultrafast microwave energy fixation method, and repeated-reaction cycling of avidin-alkaline phosphatase and a biotinylated anti-avidin antibody for signal amplification, are also briefly discussed.
利用与原癌基因ret（proto-ret）产物的激酶和跨膜结构域编码区域相对应的光生物素标记的寡脱氧核糖核酸探针，通过原位杂交检测了大鼠胚胎组织在发育过程中原癌基因ret（proto-ret）的表达。在妊娠中期（第10天和第11天），在胎盘的细胞滋养层中检测到高水平的原癌基因ret转录本，但在妊娠第14天，当胎盘发生形态变化时，滋养层细胞中不再能检测到转录本。这些结果表明，原癌基因ret表达的增加可能与滋养层细胞在特定阶段的增殖和/或分化有关。还简要讨论了通过引入超快微波能量固定方法以及抗生物素蛋白-碱性磷酸酶和生物素化抗抗生物素蛋白抗体的重复反应循环进行信号放大来改进原位杂交技术。
