Abstract
Purpose::
To study the effects of hydrostatic pressure (HP) on optic nerve head (ONH) astrocytes by focusing on gap junctions built of connexin-43 (Cx43), which form a functional syncytium allowing communication and control of ionic and metabolic homeostasis of retinal ganglion cells (RGCs) axon.
Methods::
We examined gap junction intercellular communication (GJIC) by scrape loading assays in human ONH astrocytes exposed to hydrostatic (HP) or ambient pressure (CP) in vitro. Immunostaining, immunoprecipitation and immunoblots were used to detect Cx43 distribution and phosphorylation in astrocytes exposed to HP with/without EGF receptor (EGFR) tyrosine kinase inhibitors AG1478 and AG82 and MAPK inhibitors U0126, PD98059 and SB203580.
Results::
Upon exposure to HP, astrocytes decrease GJIC and exhibit altered cellular localization and phosphorylation of Cx43. Inhibition of EGFR blocked the effects of HP on GJIC and HP-induced Cx43 tyrosine phosphorylation. Inhibitors of MAPK- ERK1/2 and -p38 caused partial closure of GJIC under CP and HP, which was maintained for 6h. Inhibition of Big Mitogen-Activated Kinase 1/ERK5 (BMK1/ERK5) caused partial closure under CP and HP followed by full recovery after 6h. Inhibition of MAPK did not affect the HP-induced increase in Cx43 serine 279/282 phosphorylation.
Conclusions::
Activation of the EGFR pathway in response to HP leads to decrease of GJIC via tyrosine phosphorylation of Cx43 in ONH astrocytes. In glaucoma under conditions of elevated intraocular pressure (IOP), astrocytes may lose GJIC altering the homeostasis of RGC axons, adopting the reactive phenotype, contributing to glaucomatous neuropathy.
Keywords: astrocytes: optic nerve head • signal transduction • gap junctions/coupling