Figure 7.
EtOH-stimulated cAMP-dependent protein kinase (PKA) activity requires the activation of cGMP-dependent protein kinase (PKG) in BBECs. Cells were treated with media only (white bars), 100 mmol/L of EtOH (black bars), or the combination of both 1 nmol/L of 8Br-cAMP and 1 nmol/L of 8Br-cGMP (gray bars), for 1 hour in the presence or absence of a 1-hour pretreatment with either 1 μmol/L of KT5720, 0.1 μmol/L of 8-pCPT-cGMPS, or media only and PKA activity assayed. EtOH or the combination of cyclic nucleotides stimulated a significant increase in PKA activity. EtOH-stimulated PKA activity was blocked by pretreatment of BBECs with 8-pCPT-cGMPS. PKA was expressed as fold increase over baseline media control cells. Bars represent SEM of separate experiments performed in triplicate (n = 9). Significance (P ≤ 0.05) indicated by an asterisk.