Battle of the kinases: Integration of adrenal responses to cAMP, DG and Ca2+at the level of steroidogenic cytochromes P450 and 3ßHSD expression in H295R cells.

Abstract

While ACTH receptors (activating the protein kinase A pathway) are expressed throughout the human/bovine/ovine zona glomerulosa (zg) and zona fasciculata (zf), there are clear zonal differences in AII Type-1 receptor levels (activating protein kinase C/Ca²⁺), as well as resting membrane potential. Thus zg is most responsive to AII and K⁺ (Ca²⁺ signalling), while zf is less responsive to AII with no K⁺ response. Zonal function in turn requires differential expression of CYP17/3ßHSD and CYP11B2/CYP11B1. We have used the H295R cell to study how differential activation of kinase A, kinase C and Ca²⁺/calmodulin (CaM) kinases may alter the relative expression of the steroidogenic P450s and 3ßHSDII. While CYP11A, CYP17, 3ßHSDII, CYP21, and CYP11B1 are all induced by increases in cAMP, studies with TPA alone or in combination with forskolin reveal subsets of steroidogenic enzymes regulated either positively (CYP21, 3ßHSDII) or negatively (CYP17, CYP11A) by protein kinase C. Thus adrenal 3ßHSDII and CYP21 expression is high in zg and zf, but CYP17 is not expressed in the zg where AII activation of kinase C is highest. In turn both K⁺ and AII-induced elevation of Ca²⁺ strongly induces CYP11B2 but not CYP11B1, consistent with preferential expression of CYP11B2 in the zg. We conclude that differential signaling through kinase C and CaM kinases in addition to kinase A underlies zonal differences in both the early and late pathways involved in steroid hormone production within the adrenocortical zones.