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Cellular force assay detects altered contractility caused by a nephritis‐associated mutation in nonmuscle myosin IIA
Author(s) -
Fukuda Shota P.,
Matsui Tsubasa S.,
Ichikawa Takafumi,
Furukawa Taichi,
Kioka Noriyuki,
Fukushima Shuichiro,
Deguchi Shinji
Publication year - 2017
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/dgd.12379
Subject(s) - biology , contractility , myosin , microbiology and biotechnology , endogeny , pathogenesis , regulator , downregulation and upregulation , mutation , hek 293 cells , genetics , gene , immunology , endocrinology
Recent progress in understanding the essential roles of mechanical forces in regulating various cellular processes expands the field of biology to one where interdisciplinary approaches with engineering techniques become indispensable. Contractile forces or contractility—inherently present in proliferative cells due to the activity of ubiquitous nonmuscle myosin II ( NMII )—are one of such mechano‐regulators, but because NMII works downstream of diverse signaling pathways, it is often difficult to predict how the inherent cellular forces change upon perturbations to particular molecules. Here, we determine whether the contractility of individual cells is upregulated or downregulated based on an assay analyzing specific deformations of silicone gel substrates. We focus on the effect of mutations in the human MYH 9 gene that encodes NMIIA , which have been implicated in the pathogenesis of various diseases including nephritis. Our assay equipped with a high‐throughput data analysis capability reveals that a point mutation of E1841K but not I1816V significantly reduces the magnitude of the endogenous forces of human embryonic kidney ( HEK 293) cells. Given the increasingly recognized roles of the endogenous forces as a critical mechano‐regulator as well as that no apparent morphological changes were induced to cells even by introducing the mutations, our findings suggest a possibility that the detected reduction in the force magnitude at the individual cellular level may underlie the pathogenesis of the kidney disease.