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KMID : 0614020080230010053
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Journal of Pharmaceutical Sciences (C.N.U.)
2008 Volume.23 No. 1 p.53 ~ p.58
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Knock-Down of Type 2 Inositol 1,4,5-Trisphosphate Receptor Using shRNA Expressing Adenovirus in Rat Ventricular Myocytes
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Son Min-Jeong
Lee Sun-woo
Woo Sun-Hee
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Abstract
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Inositol 1,4,5-trisphosphate(IP3) is a second messenger that induces the release of Ca2+ from the endoplasmic/sarcoplasmic reticulum. The IP3 receptor(IP3R)-mediated signaling pathway appears to be ubiquitous and is involved in many cellular processes including cell division, fertilization and neuronal function. Although cardiac myocytes have the IP3R, its pathophysiological role has not been clearly understood because of limited selectivity of currently available pharmacological blockers. Here we show expression of type 2 IP3R(IP3R2) at the peri-nuclear region in adult rat ventricular myocytes using western blotting and immunostaining, and successful construction of IP3R2 shRNA-expressing adenovirus to knock down the protein. The expression and localization of the IP3R2 were eliminated by 6 h-infection of the shRNA-expressing adenovirus (150 multiplicity of infection) within 2 days in primary cultured adult rat ventricular myocytes. This study provides a reliable tool for probing the role of IP3R2 in intact cardiac myocytes.
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KEYWORD
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inositol 1 , 4 , 5-triphosphate receptor (IP₃R), knock down, adult ventricular myocytes
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