TY - JOUR
T1 - Receptor-mediated endocytosis generates nanomechanical force reflective of ligand identity and cellular property
AU - Zhang, Xiao
AU - Ren, Juan
AU - Wang, Jingren
AU - Li, Shixie
AU - Zou, Qingze
AU - Gao, Nan
N1 - Funding Information:
Directorate for Biological Sciences BIO/IDBR, Grant numbers: 1353890, CMMI-1200557; National Cancer Institute, Grant number: CA178599; American Cancer Society, Grant number: RSG-15-060-01-TBE; National Institute of Diabetes and Digestive and Kidney Diseases, Grant number: DK102934
Funding Information:
This work is supported by a National Science Foundation (NSF) grant BIO/IDBR 1353890, a Charles and Johanna Busch Memorial grant to Q.Z. and N.G; an NSF grant (CMMI-1200557) to Q.Z.; National Institutes of Health (NIH) grants (R01DK102934, R21CA178599), a Research Scholar Grant (RSG-15-060-01-TBE) from the American Cancer Society, and an Initiative for Multidisciplinary Research Teams (IMRT) award to N.G.
Publisher Copyright:
© 2017 Wiley Periodicals, Inc.
PY - 2018/8
Y1 - 2018/8
N2 - Whether environmental (thermal, chemical, and nutrient) signals generate quantifiable, nanoscale, mechanophysical changes in the cellular plasma membrane has not been well elucidated. Assessment of such mechanophysical properties of plasma membrane may shed lights on fundamental cellular process. Atomic force microscopic (AFM) measurement of the mechanical properties of live cells was hampered by the difficulty in accounting for the effects of the cantilever motion and the associated hydrodynamic force on the mechanical measurement. These challenges have been addressed in our recently developed control-based AFM nanomechanical measurement protocol, which enables a fast, noninvasive, broadband measurement of the real-time changes in plasma membrane elasticity in live cells. Here we show using this newly developed AFM platform that the plasma membrane of live mammalian cells exhibits a constant and quantifiable nanomechanical property, the membrane elasticity. This mechanical property sensitively changes in response to environmental factors, such as the thermal, chemical, and growth factor stimuli. We demonstrate that different chemical inhibitors of endocytosis elicit distinct changes in plasma membrane elastic modulus reflecting their specific molecular actions on the lipid configuration or the endocytic machinery. Interestingly, two different growth factors, EGF and Wnt3a, elicited distinct elastic force profiles revealed by AFM at the plasma membrane during receptor-mediated endocytosis. By applying this platform to genetically modified cells, we uncovered a previously unknown contribution of Cdc42, a key component of the cellular trafficking network, to EGF-stimulated endocytosis at plasma membrane. Together, this nanomechanical AFM study establishes an important foundation that is expandable and adaptable for investigation of cellular membrane evolution in response to various key extracellular signals.
AB - Whether environmental (thermal, chemical, and nutrient) signals generate quantifiable, nanoscale, mechanophysical changes in the cellular plasma membrane has not been well elucidated. Assessment of such mechanophysical properties of plasma membrane may shed lights on fundamental cellular process. Atomic force microscopic (AFM) measurement of the mechanical properties of live cells was hampered by the difficulty in accounting for the effects of the cantilever motion and the associated hydrodynamic force on the mechanical measurement. These challenges have been addressed in our recently developed control-based AFM nanomechanical measurement protocol, which enables a fast, noninvasive, broadband measurement of the real-time changes in plasma membrane elasticity in live cells. Here we show using this newly developed AFM platform that the plasma membrane of live mammalian cells exhibits a constant and quantifiable nanomechanical property, the membrane elasticity. This mechanical property sensitively changes in response to environmental factors, such as the thermal, chemical, and growth factor stimuli. We demonstrate that different chemical inhibitors of endocytosis elicit distinct changes in plasma membrane elastic modulus reflecting their specific molecular actions on the lipid configuration or the endocytic machinery. Interestingly, two different growth factors, EGF and Wnt3a, elicited distinct elastic force profiles revealed by AFM at the plasma membrane during receptor-mediated endocytosis. By applying this platform to genetically modified cells, we uncovered a previously unknown contribution of Cdc42, a key component of the cellular trafficking network, to EGF-stimulated endocytosis at plasma membrane. Together, this nanomechanical AFM study establishes an important foundation that is expandable and adaptable for investigation of cellular membrane evolution in response to various key extracellular signals.
KW - atomic force microscope
KW - endocytosis
KW - mechanical force
KW - plasma membrane
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U2 - 10.1002/jcp.26400
DO - 10.1002/jcp.26400
M3 - Article
C2 - 29243828
AN - SCOPUS:85042547593
SN - 0021-9541
VL - 233
SP - 5908
EP - 5919
JO - Journal of Cellular Physiology
JF - Journal of Cellular Physiology
IS - 8
ER -