Expression of green fluorescent protein impairs the force-generating ability of isolated rat ventricular cardiomyocytes
Expression of green fluorescent protein impairs the force-generating ability of isolated rat ventricular cardiomyocytes
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DOI：
10.1007/s11010-005-9090-6
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发表时间：
2006-06-01
影响因子：
4.3
通讯作者：
Sugiura, Seiryo
中科院分区：
文献类型：
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作者：
Nishimura, Satoshi;Nagai, Shinya;Sugiura, Seiryo
绿色荧光蛋白（GFP）被广泛用作研究转基因表达在心脏组织中的作用的生物惰性表达标记物，但其对心肌细胞收缩功能的影响尚未得到充分评价。我们测量了表达GFP的重组腺病毒（Adv-GFP）感染前和感染后分离的大鼠心室肌细胞的收缩功能。用不含转基因的腺病毒（Adv-CMV）感染的肌细胞或未感染的肌细胞（UI）作为对照。使用碳纤维为基础的力-长度测量系统的单个心肌细胞，我们评估了收缩功能在广泛的负载条件下，包括缩短分数（%FS）和最大缩短速度（Vmax）在卸载条件下，和等距力。感染后24小时，近80%的Adv-GFP感染的心肌细胞表达GFR。我们发现，%FS和Vmax在三组之间没有差异，但是，只有Adv-GFP感染的心肌细胞的等长力显示出轻微但显著的下降（Adv-GFP：29.1 +/- 4.0 mN/mm（2）; Adv-GFP：42.8 +/- 6.2 mN/mm（2）; UI：47.1 +/- 4.8 mN/mm（2）; p = 0.03）。在高负荷条件下，分离的心肌细胞的收缩功能的评价显示受损的等长收缩力的GFP表达。Adv-GFP表达可能不是心肌组织中特定基因表达实验的理想对照。
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 GFR 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/mm(2); Adv-Null: 42.8 +/- 6.2 mN/mm(2); UI: 47.1 +/- 4.8 mN/mm(2); p = 0.03). An evaluation of the contractile function of isolated cardiomyocytes under high load conditions revealed impaired isometric contractility by GFP expression. Adv-GFP expression may not be an ideal control for specific gene expression experiments in myocardial tissue.