Stability of the contractile assembly and Ca^2+-activated tension in adenovirus infected adult cardiac myocytes

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• 作者：Rust ; Elizabeth M. ; Westfall ; Margaret V. ; Metzger ; Joseph M.
• 关键词：cardiac muscle ; contractility ; myofilament ; virus
• 刊名：Molecular and Cellular Biochemistry
• 出版年：1998
• 出版时间：1998
• 年：1998
• 卷：181
• 期：1/2
• 页码：143-155
• 全文大小：329 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Biochemistry
  Medical Biochemistry
  Oncology
  Cardiology
  
• 出版者：Springer Netherlands
• ISSN：1573-4919

文摘

Adenovirus-mediated gene transfer into adult cardiac myocytes in primaryculture is a potentially useful method to study the structure and functionof the contractile apparatus. However, the consequences of adenovirusinfection on the highly differentiated state of the cultured myocyte havenot been determined. We report here a detailed analysis of myofilamentstructure and function over time in primary culture and after adenovirusinfection. Adult rat ventricular myocytes in primary culture were infectedwith a recombinant adenovirus vector expressing either the LacZ or alkalinephosphatase reporter gene. Control and infected myocytes were collected atdays 0-7 post-isolation/infection, and myofilament isoform expression wasdetermined by SDS-PAGE and Western blot. Laser scanning densitometry showedthat the &agr;- to &bgr;-myosin heavy chain ratio, the stoichiometry of themyosin light chains and the expression of the adult troponin T isoform didnot change over time in culture or with adenovinus treatment. Importantly,examination of Ca$^2+$-activated tension in single myocytesshowed no change in the shape or position of the tension-pCa relationship inthe control and adenovirus infected myocytes during primary culture. Theseresults indicate that the structure and function of adult cardiac myocytesare stable in short term primary culture and are not affected by adenovirusinfection per se, and therefore provide the foundation for the use ofadenovirus-mediated myofilament gene transfer to study contractile apparatusstructure and function in adult cardiac myocytes.ain.