Protein kinase A activation of glucocorticoid-mediated signaling in the developing retina.

This report establishes that increasing the activity of cyclic AMP-dependent protein kinase (protein kinase A; PKA) potentiates glucocorticoid-mediated signaling in embryonic day 5.5 (E5.5) chicken retina. Expression of a glutamine synthetase-chloramphenicol acetyltransferase (CAT) fusion gene is not induced by treatment with glucocorticoid hormone in transfected E5.5 retina. However, treatment of the retina with forskolin, an activator of adenyl cyclase, or cotransfection with an expression vector encoding PKA is sufficient to render the fusion gene hormonally responsive. Similar results are obtained after forskolin treatment of E5.5 retina that have been transfected with a plasmid that contains the CAT reporter gene under transcriptional control by the thymidine kinase promoter and a 46-nucleotide enhancer with two glucocorticoid response elements (GREs). In contrast, forskolin augments but is not required to achieve glucocorticoid-inducible CAT gene expression in E5.5 retina transfected with a plasmid that contains the reporter driven by a minimal promoter with six juxtaposed GREs. Based on these results, we postulate that E5.5 retina contain glucocorticoid receptors whose signal transduction properties are enhanced by PKA. Unlike the transiently expressed glutamine synthetase fusion gene, however, activation of PKA does not render the endogenous glutamine synthetase gene glucocorticoid-inducible. Thus, its expression appears to be subject to an additional level of control in the developing retina.