The aim of this study was to investigate the hypotensive effect of the melatonin analogue, N-butanoyl-2-(2-methoxy-6H-isoindolo[2,1-a]indol-11-yl)ethanamine (IIK7), through MT(2)-melatonin receptors and the involvement of the sympathetic nervous system in this action in New Zealand rabbit eyes. The topical application of melatonin or IIK7 produced a reduction in intraocular pressure of 20.2 +/- 5.3% and 38.5 +/- 3.2% respectively. This effect was concentration-dependent; it was blocked by selective MT(2) receptor antagonists and was severely diminished after chemical sympathectomy. Immunohistochemistry and western blot analysis showed the ciliary processes as the site of this action and no co-localization of MT(2)-melatonin receptor with the sympathetic nervous system was observed. The beta-adrenergic agonists, terbutaline and salbutamol, potentiated the hypotensive effect of IIK7 reducing intraocular pressure (IOP) 41.75 +/- 4.26% and 44.7 +/- 5.6% respectively. Also, IIK7 in presence of the nonspecific protein phosphatase inhibitor okadaic acid, lowered IOP 32.2 +/- 4.5% and in presence of forskolin plus 3-isobutyl-1-methylxanthine decreased IOP in 32.2 +/- 5.47%. These data suggest that the melatonin agonist IIK7 reduces intraocular pressure by acting through MT(2)-melatonin receptors presumably decreasing aqueous humour formation. Also, in the presence of beta-adrenoceptor agonists MT(2)-melatonin receptors activity increase their ability to reduce IOP.