PMID: 11923256Mar 30, 2002Paper

Alterations in arachidonic acid release and phospholipase C-beta(1) expression in glaucomatous human ciliary muscle cells

Investigative Ophthalmology & Visual Science
Shahid HusainAta A Abdel-Latif

Abstract

Prostaglandin (PG) F(2alpha) and other Ca(2+)-mobilizing agonists, such as carbachol (CCh) and endothelin (ET)-1, induce an increase in uveoscleral outflow, in part through receptor-mediated mechanisms in the ciliary muscle. Because changes in uveoscleral outflow across the ciliary muscle could cause elevation of intraocular pressure (IOP) in patients with glaucoma, the present study was conducted to investigate the possibility that basal and agonist-induced second-messenger formation may be altered in glaucomatous human ciliary muscle (g-HCM) cells compared with normal (n)-HCM cells. Normal and glaucomatous HCM cells were cultured from donor eyes, the cells were identified based on their positive immunostaining with smooth muscle-specific anti-alpha-actin (SM), anti-SM-myosin, and anti-desmin antibodies. Activation of phospholipase A(2) (PLA(2)) was measured by the release of [(3)H] arachidonic acid (AA) into the medium, accumulation of PGE(2) was measured by radioimmunoassay, [(3)H]myo-inositol phosphate production was measured by ion-exchange chromatography, and phospholipase C (PLC)-beta(1) expression was determined by immunoblot analysis with polyclonal antibodies specific for PLC-beta(1). Homogenous primary cell cultures ...Continue Reading

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