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Signaling pathways in cytoskeletal responses to plasma membrane depolarization in corneal endothelial cells
Author(s) -
Evans Frances,
Hernández Julio A.,
Chifflet Silvia
Publication year - 2020
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.29200
Subject(s) - rhoa , microbiology and biotechnology , cyclic adenosine monophosphate , protein kinase a , biology , extracellular , rho associated protein kinase , signal transduction , mapk/erk pathway , cytoskeleton , kinase , biochemistry , cell , receptor
In previous work, we reported that plasma membrane potential depolarization (PMPD) provokes cortical F‐actin remodeling in bovine corneal endothelial (BCE) cells in culture, which eventually leads to the appearance of intercellular gaps. In kidney epithelial cells it has been shown that PMPD determines an extracellular‐signal‐regulated kinase (ERK)/Rho‐dependent increase in diphosphorylated myosin light chain (ppMLC). The present study investigated the signaling pathways involved in the response of BCE cells to PMPD. Differently to renal epithelial cells, we observed that PMPD leads to a decrease in monophosphorylated MLC (pMLC) without affecting diphosphorylated MLC. Also, that the pMLC reduction is a consequence of cyclic adenosine 3′,5′‐monophosphate (cAMP)/protein kinase A (PKA) activation. In addition, we found evidence that the cAMP increase mostly depends on soluble adenylyl cyclase activity. Inhibition of this enzyme reduces the effect of PMPD on the cAMP rise, F‐actin remodeling, and pMLC decrease. No changes in phosho‐ERK were observed, although we could determine that RhoA undergoes activation. Our results suggested that active RhoA is not involved in the intercellular gap formation. Overall, the findings of this study support the view that, differently to renal epithelial cells, in BCE cells PMPD determines cytoskeletal reorganization via activation of the cAMP/PKA pathway.