TY - JOUR
T1 - Rho/ROCK mechanosensor in adipocyte stiffness and traction force generation
AU - Bouzid, Tasneem
AU - Esfahani, Amir Monemian
AU - Safa, Bahareh Tajvidi
AU - Kim, Eunju
AU - Saraswathi, Viswanathan
AU - Kim, Jason K.
AU - Yang, Ruiguo
AU - Lim, Jung Yul
N1 - Funding Information:
Funding supports from National Science Foundation Graduate Research Fellowship (GRFP 1610400 , T.B.); Department of Veterans Affairs (V.S.); National Institutes of Health ( 5U2C-DK093000 , J.K.K.); National Science Foundation ( 1826135 ; 1936065 , R.Y.); National Institute of General Medical Sciences Nebraska Center for Integrated Biomolecular Communication ( P20GM113126 , R.Y.); Nebraska Collaborative Initiative (R.Y.); National Institute of General Medical Sciences Nebraska Center for the Prevention of Obesity Diseases ( P20GM104320 , J.Y.L.); and Nebraska Collaborative Initiative (J.Y.L.).
Publisher Copyright:
© 2022 The Authors
PY - 2022/5/28
Y1 - 2022/5/28
N2 - It is increasingly recognized that interaction of adipose cells with extracellular mechanophysical milieus may play a role in regulating adipogenesis and differentiated adipocyte function and such interaction can be mediated by the mechanics of adipose cells. We measured the stiffness and traction force of adipose cells and examined the role of Rho/ROCK, the upstream effector of actin cytoskeletal contractility, in affecting these mechanical properties. Cellular Young's modulus obtained from atomic force microscopy (AFM) was significantly reduced by ROCK inhibitor (Y-27632) but elevated by Rho activator (CN01), for both preadipocytes and differentiated adipocytes. Immunofluorescent imaging suggested this could be attributed to the changes in Rho/ROCK-induced stressed actin filament formation. AFM also confirmed that differentiated adipocytes had higher stiffness than preadipocytes. On the other hand, traction force microscopy (TFM) revealed differentiated adipocytes exerted lower traction forces than preadipocytes. Traction forces of both preadipocytes and adipocytes were decreased by ROCK inhibition, but not significantly altered by Rho activation. Notably, an increasing trend of traction force with respect to cell spreading area was detected, and this trend was substantially amplified by Rho activation. Such traction force-cell area correlation was an order-of-magnitude smaller for differentiated adipocytes relative to preadipocytes, potentially due to disrupted force transmission through cytoskeleton-focal adhesion linkage by lipid droplets. Our work provides new data evidencing the Rho/ROCK control in adipose cell mechanics, laying the groundwork for adipocyte mechanotransduction studies on adipogenesis and adipose tissue remodeling.
AB - It is increasingly recognized that interaction of adipose cells with extracellular mechanophysical milieus may play a role in regulating adipogenesis and differentiated adipocyte function and such interaction can be mediated by the mechanics of adipose cells. We measured the stiffness and traction force of adipose cells and examined the role of Rho/ROCK, the upstream effector of actin cytoskeletal contractility, in affecting these mechanical properties. Cellular Young's modulus obtained from atomic force microscopy (AFM) was significantly reduced by ROCK inhibitor (Y-27632) but elevated by Rho activator (CN01), for both preadipocytes and differentiated adipocytes. Immunofluorescent imaging suggested this could be attributed to the changes in Rho/ROCK-induced stressed actin filament formation. AFM also confirmed that differentiated adipocytes had higher stiffness than preadipocytes. On the other hand, traction force microscopy (TFM) revealed differentiated adipocytes exerted lower traction forces than preadipocytes. Traction forces of both preadipocytes and adipocytes were decreased by ROCK inhibition, but not significantly altered by Rho activation. Notably, an increasing trend of traction force with respect to cell spreading area was detected, and this trend was substantially amplified by Rho activation. Such traction force-cell area correlation was an order-of-magnitude smaller for differentiated adipocytes relative to preadipocytes, potentially due to disrupted force transmission through cytoskeleton-focal adhesion linkage by lipid droplets. Our work provides new data evidencing the Rho/ROCK control in adipose cell mechanics, laying the groundwork for adipocyte mechanotransduction studies on adipogenesis and adipose tissue remodeling.
KW - Adipocyte
KW - Mechanotransduction
KW - Rho/ROCK
KW - Traction force
KW - Young's modulus
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U2 - 10.1016/j.bbrc.2022.03.078
DO - 10.1016/j.bbrc.2022.03.078
M3 - Article
C2 - 35339750
AN - SCOPUS:85126861318
SN - 0006-291X
VL - 606
SP - 42
EP - 48
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
ER -