Photomechanical movements of cultured embryonic photoreceptors: regulation by exogenous neuromodulators and by a regulable source of endogenous dopamine

Abstract

In the retina of nonmammalian vertebrates, light regulates photoreceptor morphology by causing rod photoreceptor elongation and cone photoreceptor contraction. The opposite photomechanical movements occur in the dark, and proceed with a circadian rhythm in many species in vivo. Using dissociated cultures of embryonic chick retina cells, we have recently demonstrated that photoreceptor cells that differentiate in vitro acquire the capacity of responding to light/dark cycles with photomechanical movements (Stenkamp and Adler, 1993). Here we report that the putative neuromodulators melatonin and dopamine can mimic the effects of darkness and light, respectively, on in vitro photomechanical movement. Pharmacological studies showed that dopamine appears to function by means of a D2-type receptor negatively coupled to adenylate cyclase. The effects of light on the cultured photoreceptors were inhibited by dopamine D2 receptor antagonists, and were attenuated by the dopaminergic neurotoxin 6-hydroxydopamine and by the dopamine synthesis inhibitor alpha-methyl-p-tyrosine. The possible existence of an endogenous source of dopamine in the cultures was also suggested by the presence of tyrosine hydroxylase-like immunoreactivity, and of an Na(+)-dependent mechanism for the accumulation of 3H-dopamine, which was predominantly associated with nonphotoreceptor cells. Additionally, 3H-dopamine release occurred in vitro through a Ca(2+)-dependent mechanism, as well as through reverse function of a nomifensine-sensitive dopamine transporter. Both of these putative release mechanisms appeared to be regulated by light and by melatonin, suggesting a mechanism whereby the putative dopaminergic cells may interact with other cells present in the cultures. These studies suggest that complex paracrine neuromodulatory mechanisms can differentiate in low-density embryonic cell culture, that dopaminergic activities exist in vitro, and that they are important for mediating photomechanical movements.