Regulation of melatonin and dopamine biosynthesis in chick retina: The role of GABA

Melatonin biosynthesis in chick retina occurs as a circadian rhythm. Biosynthesis of the neurohormone is highest at night in darkness, and is suppressed by light. The role of gamma-aminobutyric acid (GABA) in the nocturnal regulation of melatonin synthesis was examined. Systemic or intravitreal administration of muscimol, a GABA-A receptor agonist, to light-exposed chicks at the beginning of the dark phase of the light/dark cycle increased retinal melatonin levels and the activity of serotonin N-acetyltransferase (NAT), a key regulatory enzyme of the melatonin biosynthetic pathway. Baclofen, a GABA-B receptor agonist, also increased NAT activity of light-exposed retinas, but muscimol was approximately 40-fold more potent than baclofen. Effects of both muscimol and baclofen on NAT activity were inhibited by GABA-A antagonists, bicuculline and picrotoxin, and the effect of baclofen was unaffected by the GABA-B selective antagonist, CGP 35348. Thus, activation of GABA-A receptors appears to be associated with increased melatonin biosynthesis. The GABA-uptake inhibitor, nipecotic acid, and the GABA-transaminase inhibitor, aminooxyacetic acid, also increased NAT activity of light-exposed retinas. The high levels of NAT activity associated with exposure to darkness were unaffected by either muscimol or baclofen, but picrotoxin and bicuculline significantly inhibited retinal NAT activity in darkness.The rate of dopamine synthesis, estimated from in situ tyrosine hydroxylase activity, was higher in light-exposed retinas than in darkness. Muscimol inhibited dopamine synthesis in light, and picrotoxin stimulated dopamine synthesis in darkness. The stimulation of melatonin synthesis by muscimol in light-exposed retinas appears to be related to inhibition of retinal dopamine neurons. The increase of NAT activity elicited by muscimol in light-exposed retinas was inhibited by administration of the dopamine receptor agonists apomorphine and quinpirole. Blocking dopamine receptors with spiperone or inhibiting dopamine biosynthesis with α-methyl-ρ tyrosine also increased NAT activity in light, and the effects of the dopamine antagonists and muscimol were not additive. The decrease of NAT activity elicited by GABA antagonists in darkness was inhibited by spiperone. Thus, GABA may indirectly regulate retinal melatonin biosynthesis, by inhibiting dopaminergic activity in retina.