期刊
JOURNAL OF CELLULAR PHYSIOLOGY
卷 235, 期 3, 页码 2947-2962出版社
WILEY
DOI: 10.1002/jcp.29200
关键词
actin cytoskeleton; cAMP; PKA; corneal endothelium; myosin light chain; plasma membrane depolarization
资金
- Agencia Nacional de Investigacion e Innovacion (ANII), Uruguay [PR_FCE_2009_1_2474]
- Comision Sectorial de Investigacion Cientifica (CSIC), Universidad de la Republica, Uruguay
- Programa de Desarrollo de las Ciencias Basicas (PEDECIBA), Uruguay
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.
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