Boer P H
University of Ottawa Heart Institute, Department of Pathology & Laboratory Medicine, Ontario, Canada.
Biochem Biophys Res Commun. 1994 Mar 15;199(2):954-61. doi: 10.1006/bbrc.1994.1322.
We assessed the temporal transcriptional activity profiles of the genes for type-B natriuretic factor, BNF, the isoform ANF, and other cardiac muscle proteins in differentiating cultures derived from multipotential mouse cell lines. P19 embryonal carcinoma cells and D3 embryonic stem cells were induced for in vitro cardiac myogenesis; RNA was isolated at regular intervals throughout the differentiation programs, and mRNAs were detected by reverse transcriptase mediated polymerase chain reactions. The transcriptional activation profiles of the ANF and BNF gene were similar, but there were quantitative differences that were best assayed by use of competitive internal DNA standards. The levels of induced BNF transcripts were highest in the P19 developmental system reaching approximately 10% of adult mouse ventricular muscle levels; those for ANF were lower, but also readily detected. The cell lines may be used to define the regulatory control elements for natriuretic factor gene expression, in stably transfected cell lines, during cardiac muscle growth.
我们评估了源自多能小鼠细胞系的分化培养物中B型利钠因子（BNF）、ANF同工型以及其他心肌蛋白基因的时间转录活性谱。诱导P19胚胎癌细胞和D3胚胎干细胞进行体外心肌生成；在整个分化过程中定期分离RNA，并通过逆转录介导的聚合酶链反应检测mRNA。ANF和BNF基因的转录激活谱相似，但存在定量差异，使用竞争性内部DNA标准进行检测最为合适。在P19发育系统中，诱导的BNF转录本水平最高，达到成年小鼠心室肌水平的约10%；ANF的水平较低，但也易于检测到。这些细胞系可用于在稳定转染的细胞系中，在心肌生长过程中确定利钠因子基因表达的调控元件。
