論文

基本情報

氏名 竹谷 浩介
氏名(カナ) タケヤ コウスケ
氏名(英語) Takeya Kosuke
所属 獣医学部 獣医学科
職名 講師
researchmap研究者コード 5000068922
researchmap機関 岡山理科大学

題名

A highly sensitive technique to measure myosin regulatory light chain phosphorylation: the first quantification in renal arterioles.

単著・共著の別

共著

著者

Takeya K, Loutzenhiser K, Shiraishi M, Loutzenhiser R, Walsh MP

概要

Phosphorylation of the 20-kDa myosin regulatory light chains (LC(20)) plays a key role in the regulation of smooth muscle contraction. The level of LC(20) phosphorylation is governed by the relative activities of myosin light chain kinase and phosphatase pathways. The regulation of these two pathways differs in different smooth muscle types and in the actions of different vasoactive stimuli. Little is known concerning the regulation of LC(20) phosphorylation in the renal microcirculation. The available pharmacological probes are often nonspecific, and current techniques to directly measure LC(20) phosphorylation are not sensitive enough for quantification in small arterioles. We describe here a novel approach to address this important issue. Using SDS-PAGE with polyacrylamide-bound Mn(2+)-phosphate-binding tag and enhanced Western blot analysis, we were able to detect LC(20) phosphorylation using as little as 5 pg (250 amol) of isolated LC(20). Phosphorylated and unphosphorylated LC(20) were detected in single isolated afferent arterioles, and LC(20) phosphorylation levels could be accurately quantified in pooled samples of three arterioles (<300 cells). The phosphorylation level of LC(20) in the afferent arteriole was 6.8 +/- 1.7% under basal conditions and increased to 34.7 +/- 5.1% and 44.6 +/- 6.6% in response to 30 mM KCl and 10(-8) M angiotensin II, respectively. The application of this technique will enable investigations of the different determinants of LC(20) phosphorylation in afferent and efferent arterioles and provide insights into the signaling pathways that regulate LC(20) phosphorylation in the renal microvasculature under physiological and pathophysiological conditions.    

発表雑誌等の名称

American Journal of Physiology Renal Physiology

出版者

294

6

開始ページ

F1487

終了ページ

F1492

発行又は発表の年月

2008/06

査読の有無

有り

招待の有無

無し

記述言語

英語

掲載種別

研究論文(学術雑誌)

ISSN

ID:DOI

10.1152/ajprenal.00060.2008

ID:NAID(CiNiiのID)

ID:PMID

18400874

URL

JGlobalID

arXiv ID

ORCIDのPut Code

DBLP ID