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The importance of light in the temporal organization of photoreceptor proteins and melatonin-producing system has been investigated for the first time in the pineal of a tropical fish. In this study, an identical experimental paradigm was followed during the four distinct phases of an annual cycle in adult carps (Catla catla) maintained either under natural photoperiod (NP) or continuous illumination (LL) or darkness (DD) for 30 days. At the end of each experiment, the pineal from fish in each experimental group was collected either at 06:00, 12:00, 18:00, or 24:00 in a daily cycle and assessed by Western blot analysis for pineal rod-like opsin, alpha-transducin, and AANAT. The same animals were also used for measurement of serum melatonin levels, and the serum as well as intra-pineal Ca(++) levels at each timepoint. The study revealed a daily rhythmicity with a peak at 12:00 h and nadir at 24:00 h in the band intensity of pineal rod-like opsin and alpha-transducin in NP fish, while the band intensities of these photo-pigment proteins remained high under LL and low under DD, irrespective of clock hour during the 24 h cycle. The band intensity of pineal AANAT, levels of serum melatonin, and both serum Ca(++) and intra-pineal Ca(++) were maximum at 24:00 h and minimum at 12:00h in NP fish, and they were significantly lower under LL and higher under DD at each point of study. The results showed loss of daily rhythm in each studied variable in both LL and DD carps, suggesting that their circadian organization is dependent on the external light-dark conditions, rather than an endogenous circadian oscillator in the pineal.

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In the present in vitro study on the pineal in carp Catla catla, specific agonist and antagonists of receptors for different neuronal signals and regulators of intra-cellular Ca(++) and cAMP were used to gather basic information on the neuronal signal transduction cascade mechanisms in the photo-induced expression of rod-like opsin and alpha-transducin-like proteins in any fish pineal. Western-blot analysis followed by quantitative analysis of respective immunoblot data for both the proteins revealed that photo-induced expression of each protein was stimulated by cholinergic (both nicotinic and muscarinic) agonists and a dopaminergic antagonist, inhibited by both cholinergic antagonists and a dopaminergic agonist, but not affected by any agonists or antagonists of adrenergic (alpha(1), alpha(2) and beta(1)) receptors. Moreover, expression of each protein was stimulated by voltage gated L type calcium channel blocker, adenylate cyclase inhibitor and phosphodiesterase activator; but suppressed by the activators of both calcium channel and adenylate cyclase, and by phosphodiesterase inhibitor. Collectively, we report for the first time that both cholinergic and dopaminergic signals play an important, though antagonistic, role in the photo-induced expression of photoreceptor proteins in the fish pineal through activation of a signal transduction pathway in which both calcium and cAMP may act as the intracellular messengers.