Inhibition of the cardiac ?-actin gene in embryonic cardiac myocytes by dominant-negative serum response factor
✍ Scribed by Zhu, Xiaolei ;McAllister, Donna ;Lough, John
- Publisher
- John Wiley and Sons
- Year
- 2003
- Tongue
- English
- Weight
- 300 KB
- Volume
- 271A
- Category
- Article
- ISSN
- 0003-276X
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
Serum response factor (SRF), a transcription factor ubiquitously involved in the processes of cellular proliferation and differentiation, has been implicated in cardiac and skeletal muscle development because of its strong expression in embryonic muscle lineages, and its necessity for the transcription of transiently transfected muscle genes that contain SRF binding sites. This study was designed to ascertain whether SRF is required for the expression of an endogenous SRF‐dependent gene during differentiation of early embryonic cardiac myocytes by introducing a dominant‐negative SRF construct via retroviral delivery. Although no effect on overt cellular differentiation was detected, semi‐quantitative RT‐PCR revealed that expression of the SRF‐dependent gene cardiac α‐actin was inhibited, whereas expression of the non‐SRF‐dependent genes glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) and cardiac troponin‐C was unaffected. No effect on myocyte proliferation was detected. Curiously, immunohistochemical localization of SRF protein suggested that whereas endogenous SRF was homogeneously dispersed throughout the cytoplasm and nucleus, the dominant‐negative SRF protein was concentrated in the nucleus. These results extend previous findings using transiently transfected genes to the endogenous level, indicating that SRF is required for the full expression of muscle genes that contain SRF binding sites during cardiac myocyte differentiation. Anat Rec Part A 271A:315–321, 2003. © 2003 Wiley‐Liss, Inc.