Regulation of tyrosine hydroxylase activity in retinal cell suspensions: Effects of potassium and 8-bromo cyclic AMP

Abstract

Tyrosine hydroxylase (TH) contained in dopamine (DA) neurons of rat retina is activated in vivo as a consequence of photic stimulation. Experiments were conducted to test the effects of changes of membrane potential and of cyclic AMP-dependent protein phosphorylation on TH activity of these retinal neurons. Retinas were dissociated into suspensions of apparently viable cells to test the direct effects of pharmacological manipulations of TH activity in the absence of trans-synaptic influences. To test the effects of changes of membrane potential on TH activity we examined the effects of a depolarizing agent, potassium. Incubation of cell suspensions in Ringer's solution containing a depolarizing concentration of K + (52 mM) resulted in a significant increase of TH activity, suggesting that membrane depolarization may trigger a series of molecular events that leads to TH activation. Incubation of cell suspensions in the presence of 8-bromo cyclic AMP, a cyclic AMP analog that is known to activate cyclic AMP-dependent processes following extra-cellular application, resulted in a significant activation of TH that was comparable to that produced in vitro by cyclic AMP-dependent protein phosphorylation. These data support the hypothesis that membrane potential plays a role in the regulation of TH activity, and indicate that cyclic AMP-dependent phosphorylation can activate retinal TH in situ . The apparent viability of the retinal cells in suspension suggests that this preparation may be useful for studying synaptic regulatory mechanisms.