Expression of green fluorescent protein impairs the force-generating ability of isolated rat ventricular cardiomyocytes

Molecular and Cellular Biochemistry
Satoshi NishimuraS Sugiura

Abstract

Green fluorescent protein (GFP) is widely used as a biologically inert expression marker for studying the effects of transgene expression in heart tissue, but its influence on the contractile function of cardiomyocytes has not yet been fully evaluated. We measured the contractile function of isolated rat ventricular myocytes before and after infection with a recombinant adenovirus expressing GFP (Adv-GFP). Myocytes infected with a non-transgene-containing adenovirus (Adv-Null) or uninfected myocytes (UI) served as controls. Using a carbon-fiber-based force-length measurement system for single cardiomyocytes, we evaluated the contractile function over a wide range of loading conditions including the shortening fraction (%FS) and maximal shortening velocity (Vmax) under the unloaded condition, and isometric force. At 24 hours after infection, nearly 80% of the Adv-GFP-infected myocytes expressed GFP. We found that the %FS and Vmax did not differ among the three groups, however, the isometric force showed a mild, but significant, decrease only in Adv-GFP myocytes (Adv-GFP: 29.1 +/- 4.0 mN/mm2; Adv-Null: 42.8 +/- 6.2 mN/mm2; UI: 47.1 +/- 4.8 mN/mm2; p = 0.03). An evaluation of the contractile function of isolated cardiomyocytes und...Continue Reading

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Citations

Aug 7, 2007·American Journal of Physiology. Heart and Circulatory Physiology·Rajesh B SekarLeslie Tung
Nov 19, 2014·Evidence-based Child Health : a Cochrane Review Journal·Olabisi OduwoleEkong E Udoh
Mar 1, 2008·Nature Methods·Daniel I ResnicowLeslie A Leinwand
Aug 25, 2011·Journal of Experimental Botany·Rosemary G White, Deborah A Barton
Jun 13, 2008·Pflügers Archiv : European journal of physiology·Stephanie ReisKornelia Jaquet
Jun 19, 2010·Journal of Cardiovascular Translational Research·Sivaraj SivaramakrishnanJames A Spudich
May 13, 2008·Progress in Biophysics and Molecular Biology·Satoshi NishimuraSeiryo Sugiura
Sep 20, 2011·Journal of Molecular and Cellular Cardiology·F S KorteMichael Regnier

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