Laminin regulates mouse embryonic stem cell migration: involvement of Epac1/Rap1 and Rac1/cdc42

被引:28
作者
Suh, Han Na [1 ]
Han, Ho Jae [1 ]
机构
[1] Chonnam Natl Univ, Coll Vet Med, Dept Vet Physiol, Biotherapy Human Resources Ctr BK 21, Kwangju 500757, South Korea
来源
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY | 2010年 / 298卷 / 05期
基金
新加坡国家研究基金会;
关键词
cAMP/Epac1/Rap1; Rac1/cdc42/p21-activated kinase; E-cadherin; FOCAL ADHESION KINASE; RAC; INTEGRINS; RAP1; EXPRESSION; BINDING; DIFFERENTIATION; RECEPTOR; EMBRYOGENESIS; PLURIPOTENCY;
D O I
10.1152/ajpcell.00496.2009
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Suh HN, Han HJ. Laminin regulates mouse embryonic stem cell migration: involvement of Epac1/Rap1 and Rac1/cdc42. Am J Physiol Cell Physiol 298: C1159-C1169, 2010. First published January 20, 2010; doi:10.1152/ajpcell.00496.2009.-Laminin is the first extracellular matrix (ECM) component to be expressed in the developing mammalian embryo. However, the roles of laminin or the related signal pathways are not well known in mouse embryonic stem cells (mESCs). Presently, we examined the effect of laminin on mESC migration. Laminin (10 mu g/ml) decreased cell aggregation, whereas migration was increased. Laminin bound alpha 6 beta 1 integrin and laminin receptor 1 (LR1), decreasing their mRNA levels. Laminin increased focal adhesion kinase (FAK) and paxillin phosphorylation, cAMP intracellular concentration, and the protein levels of exchange factor directly activated by cAMP (Epac1) and Rap1. These increases were completely blocked by alpha 6 beta 1 integrin and LR1 neutralizing antibody, indicating that laminin-bound LR1 assists laminin-induced alpha 6 beta 1 integrin activity and initiates signal. As a downstream signal molecule, laminin activated small G protein such as Rac1/cdc42 and its effector protein p21-activated kinase (PAK). Subsequently, laminin stimulated E-cadherin complex disruption. Inhibition of each pathway such as those for alpha 6 beta 1 integrin and LR1, FAK, Rap1, and PAK1 blocked laminin-induced migration. We conclude that laminin binds both alpha 6 beta 1 integrin and LR1 and induces signaling FAK/paxillin and cAMP/Epac1/Rap1. These signaling merge at Rac1/cdc42 subsequently activate PAK1. Activated PAK1 enhances E-cadherin complex disruption and finally increases mESCs migration.
引用
收藏
页码:C1159 / C1169
页数:11
相关论文
共 66 条
[1]   RAC1 regulates adherens junctions through endocytosis of E-cadherin [J].
Akhtar, N ;
Hotchin, NA .
MOLECULAR BIOLOGY OF THE CELL, 2001, 12 (04) :847-862
[2]  
Alenghat Francis J, 2002, Sci STKE, V2002, ppe6, DOI 10.1126/stke.2002.119.pe6
[3]   Mechanical Control of cAMP Signaling Through Integrins Is Mediated by the Heterotrimeric Gαs Protein [J].
Alenghat, Francis J. ;
Tytell, Jessica D. ;
Thodeti, Charles K. ;
Derrien, Alexandrine ;
Ingber, Donald F. .
JOURNAL OF CELLULAR BIOCHEMISTRY, 2009, 106 (04) :529-538
[4]   β1 integrin deficiency impairs migration and differentiation of mouse embryonic stem cell derived neurons [J].
Andressen, C ;
Arnhold, S ;
Puschmann, M ;
Bloch, W ;
Hescheler, J ;
Fässler, R ;
Addicks, K .
NEUROSCIENCE LETTERS, 1998, 251 (03) :165-168
[5]  
Ardini E, 1997, J BIOL CHEM, V272, P2342
[6]   What mouse mutants teach us about extracellular matrix function [J].
Aszodi, A. ;
Legate, Kyle R. ;
Nakchbandi, I. ;
Faessler, R. .
ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY, 2006, 22 :591-621
[7]   A simplified laminin nomenclature [J].
Aumailley, M ;
Bruckner-Tuderman, L ;
Carter, WG ;
Deutzmann, R ;
Edgar, D ;
Ekblom, P ;
Engel, J ;
Engvall, E ;
Hohenester, E ;
Jones, JCR ;
Kleinman, HK ;
Marinkovich, MP ;
Martin, GR ;
Mayer, U ;
Meneguzzi, G ;
Miner, JH ;
Miyazaki, K ;
Patarroyo, M ;
Paulsson, M ;
Quaranta, V ;
Sanes, JR ;
Sasaki, T ;
Sekiguchi, K ;
Sorokin, LM ;
Talts, JF ;
Tryggvason, K ;
Uitto, J ;
Virtanen, I ;
von der Mark, K ;
Wewer, UM ;
Yamada, Y ;
Yurchenco, PD .
MATRIX BIOLOGY, 2005, 24 (05) :326-332
[8]   Rap1 signalling: Adhering to new models [J].
Bos, JL ;
de Rooij, J ;
Reedquist, KA .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2001, 2 (05) :369-377
[9]   Linking Rap to cell adhesion [J].
Bos, JL .
CURRENT OPINION IN CELL BIOLOGY, 2005, 17 (02) :123-128
[10]   Feeder-free culture of human embryonic stem cells in conditioned medium for efficient genetic modification [J].
Braam, Stefan R. ;
Denning, Chris ;
Matsa, Elena ;
Young, Lorraine E. ;
Passier, Robert ;
Mummery, Christine L. .
NATURE PROTOCOLS, 2008, 3 (09) :1435-1443