Immunofluorescent detection of the activation of the small GTPase Rac1 in mouse skeletal muscle fibers

被引:12
作者
Takenaka, Nobuyuki [1 ]
Nihata, Yuma [1 ]
Satoh, Takaya [1 ]
机构
[1] Osaka Prefecture Univ, Grad Sch Sci, Dept Biol Sci, Cell Biol Lab,Naka Ku, Sakai, Osaka 5998531, Japan
关键词
Activated form; Immunofluorescent microscopy; Rac1; Skeletal muscle; Small GTPase; Subcellular localization; TRANSLOCATION;
D O I
10.1016/j.ab.2014.09.013
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The small GTPase Rac1 acts as a molecular switch of intracellular signaling in mammals. For understanding the regulatory mechanism, it is important to identify subcellular locations in which Rac1 is activated following multiple extracellular stimuli. However, it is difficult to detect Rac1 activation in situ in animal tissues, and thus a novel method is highly desirable. Here, we report a simple method to visualize the activation of endogenous Rac1 in mouse skeletal muscle fibers. In this assay, specific interaction between activated Rac1 and a binding polypeptide is detected by immunofluorescent microscopy. This approach is readily applicable to other small GTPases. (C) 2014 Elsevier Inc. All rights reserved.
引用
收藏
页码:5 / 7
页数:3
相关论文
共 13 条
[1]   Rac1 signalling towards GLUT4/glucose uptake in skeletal muscle [J].
Chiu, Tim T. ;
Jensen, Thomas E. ;
Sylow, Lykke ;
Richter, Erik A. ;
Klip, Amira .
CELLULAR SIGNALLING, 2011, 23 (10) :1546-1554
[2]   The GLUT4 glucose transporter [J].
Huang, Shaohui ;
Czech, Michael P. .
CELL METABOLISM, 2007, 5 (04) :237-252
[3]   Role of the guanine nucleotide exchange factor Ost in negative regulation of receptor endocytosis by the small GTPase Rac1* [J].
Ieguchi, Katsuaki ;
Ueda, Shuji ;
Kataoka, Tohru ;
Satoh, Takaya .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (32) :23296-23305
[4]   Impaired vascular development in the yolk sac and allantois in mice lacking RA-GEF-1 [J].
Kanemura, Hoshimi ;
Satoh, Takaya ;
Bilasy, Shymaa E. ;
Ueda, Shuji ;
Hirashima, Masanori ;
Kataoka, Tohru .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2009, 387 (04) :754-759
[5]   Cell migration: Rho GTPases lead the way [J].
Raftopoulou, M ;
Hall, A .
DEVELOPMENTAL BIOLOGY, 2004, 265 (01) :23-32
[6]  
Satoh Takaya, 2014, Small GTPases, V5, pe28102, DOI 10.4161/sgtp.28102
[7]   Small GTP-binding proteins [J].
Takai, Y ;
Sasaki, T ;
Matozaki, T .
PHYSIOLOGICAL REVIEWS, 2001, 81 (01) :153-208
[8]   A critical role of the small GTPase Rac1 in Akt2-mediated GLUT4 translocation in mouse skeletal muscle [J].
Takenaka, Nobuyuki ;
Izawa, Rumi ;
Wu, Junyuan ;
Kitagawa, Kaho ;
Nihata, Yuma ;
Hosooka, Tetsuya ;
Noguchi, Tetsuya ;
Ogawa, Wataru ;
Aiba, Atsu ;
Satoh, Takaya .
FEBS JOURNAL, 2014, 281 (05) :1493-1504
[9]   A new Rac target POSH is an SHS-containing scaffold protein involved in the JNK and NF-κB signalling pathways [J].
Tapon, N ;
Nagata, K ;
Lamarche, N ;
Hall, A .
EMBO JOURNAL, 1998, 17 (05) :1395-1404
[10]   Role of the Sec14-like domain of Dbl family exchange factors in the regulation of Rho family GTPases in different subcellular sites [J].
Ueda, S ;
Kataoka, T ;
Satoh, T .
CELLULAR SIGNALLING, 2004, 16 (08) :899-906