Metabolism of phosphoinositides and inositol polyphosphates in rabbit corneal epithelium

Abstract

The synthesis of phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2) was studied in the rabbit cornea using 32P and [3H]myo-inositol as precursors. Also, the formation of water-soluble products of the degradation of inositol lipids, inositol-1-phosphate, -1,4-bisphosphate and -1,4,5-trisphosphate, was shown. Corneal epithelium displayed the most active inositol lipid metabolism and endothelium the least when the cornea layers were separately incubated (stroma had intermediate values). In corneal epithelium incubated with [3H]myo-inositol, tritiated glycerophosphorylinositol and water-soluble inositol phosphates were formed. 10 mM LiCl promotes an increase in the inositol phosphates consistent with an inhibitory effect on inositol phosphatase. Cell-free preparations of epithelium incubated with [gamma-32P]ATP detected the presence of diacylglycerol kinase, PI kinase and PIP kinase. Endogenous PI was rapidly phosphorylated to PIP within 1 min of incubation, whereas PIP was phosphorylated more slowly. In conclusion, the components of the inositol lipid cycle are present in the cornea, particularly in the epithelium. It is proposed that the control of these pathways may be involved in the transduction of cell signals through the plasma membrane, intracellular calcium ionization and epithelial cell proliferation and differentiation, particularly in wound healing.