Abstract:
:We examined the transcriptional activity profile of the gene for atrial natriuretic factor (ANF) in mouse embryonal carcinoma P19 cells which had been induced for in vitro cardiac myogenesis. Differentiation was assessed visually, by the degree of spontaneous beating activity, and by the appearance of striated muscle structures detected by immunofluorescence with a myosin heavy chain antibody. Northern blot analysis of RNA isolated at regular intervals throughout the differentiation program revealed abundant cardiac alpha-actin transcripts beginning at Day 6, reaching maximum levels during Days 7 to 8 and declining to low levels by Days 12 to 15. Throughout this period, the transcriptional profile of the ANF gene was similar to that of alpha-actin but at lower levels; thus, in vivo stages of abundant ANF and structural muscle gene transcription were not reached and these gene expression states appear to be uncoupled. Using the more sensitive assay of reverse transcriptase-mediated polymerase chain reactions, we observed the presence of ANF transcripts even in small samples of muscle-induced P19 cells and not in neuron-induced or undifferentiated P19 cells. Induced ANF transcript levels reached about 5-10% that found in adult atrium muscle tissue. ANF gene activity was further corroborated by nuclear transcriptional run-on assays. The P19 stem cell model system will be of value in the study of early events during cardiac muscle commitment and differentiation.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Boer PHdoi
10.1006/excr.1993.1210subject
Has Abstractpub_date
1993-08-01 00:00:00pages
421-9issue
2eissn
0014-4827issn
1090-2422pii
S0014-4827(83)71210-3journal_volume
207pub_type
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journal_title:Experimental cell research
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pub_type: 杂志文章
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journal_title:Experimental cell research
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pub_type: 杂志文章,评审
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