Neural regulation of dark-induced abundance of arylalkylamine N-acetyltransferase (AANAT) and melatonin in the carp (Catla catla) pineal: an in vitro study.

In all the vertebrates, synthesis of melatonin and its rhythm-generating enzyme arylalkylamine N-acetyltransferase (AANAT) reaches its peak in the pineal during the night in a daily light-dark cycle, but the role of different neuronal signals in their regulation were unknown for any fish. Hence, the authors used specific agonist and antagonists of receptors for different neuronal signals and regulators of intracellular calcium (Ca(2+)) and adenosine 3',5'-cyclic monophosphate (cAMP) in vitro to study their effects on the abundance of AANAT and titer of melatonin in the carp (Catla catla) pineal. Western blot analysis followed by quantitative analysis of respective immunoblot data for AANAT protein, radioimmunoassay of melatonin, and spectrophotometric analysis of Ca(2+) in the pineal revealed stimulatory effects of both adrenergic (α(1) and β(1)) and dopaminergic (D(1)) agonists and cholinergic (both nicotinic and muscarinic) antagonists, inhibition by both adrenergic and dopaminergic antagonists and cholinergic agonists, but independent of the influence of any agonists or antagonists of α(2)-adrenergic receptors.

Importance of light in temporal organization of photoreceptor proteins and melatonin-producing system in the pineal of carp Catla catla.

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.

Neuronal regulation of photo-induced pineal photoreceptor proteins in carp Catla catla.

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.

Photoreceptor proteins and melatonin rhythm generating AANAT in the carp pineal: Temporal organization and correlation with natural photo-thermal cues.

We studied temporal organization of both the photoreceptor (rod-like opsin, alpha subunit of the G protein transducin or alpha-TD) and melatonin generating (AANAT) proteins in the same pineal of a tropical surface dwelling free-living carp Catla catla, and analyzed possible correlation between them as well as with natural photo-thermal variables in an annual cycle. The pineal from individual fish was collected at four different time points (06.00 h, 12.

Localization and dynamics of Mel(1a) melatonin receptor in the ovary of carp Catla catla in relation to serum melatonin levels.

We studied the localization, sub-cellular distribution and daily rhythms of a 37 kDa melatonin receptor (Mel(1a)R) in the ovary to assess its temporal relationship with the serum melatonin levels in four different reproductive phases in carp Catla catla. Our immunocytochemical study accompanied by Western blot analysis of Mel(1a)R in the ovary revealed that the expression of this 37-kDa protein was greater in the membrane fraction than in the cytosol. Ovarian Mel(1a)R protein peaked at midnight and fell at midday in each reproductive phase.

Influence of serotonin on the action of melatonin in MIH-induced meiotic resumption in the oocytes of carp Catla catla.

The influences of serotonin (5-hydroxytryptamine) on the action of melatonin (N-acetyl-5-methoxytryptamine) in MIH (maturation inducing hormone)-induced meiotic resumption were evaluated in the oocytes of carp Catla catla using an in vitro model. Oocytes from gravid female carp were isolated and incubated separately in Medium 199 containing either (a) only melatonin (MEL; 100 pg/mL), or (b) only serotonin (SER; 100 pg/mL), or (c) only MIH (1 microg/mL), or (d) MEL and MIH (e) or MEL (4 h before) and MIH, or (f) MEL and SER, (g) or SER and MIH, or (h) SER (4 h before) and MIH, or (i) luzindole (L-antagonist of MEL receptors; 10 microM) and MEL, or (j) MEL, L and MIH, or (k) MEL (4 h before), L and MIH, or (l) metoclopramide hydrochloride (M-antagonist of SER receptors; 10 microM) and SER, or (m) M, MEL, SER, or (n) M, SER and MIH, or (o) M, SER (4 h before) and MIH, or (p) M, MEL SER and MIH, or (q) MEL, L, SER and M, or (r) MEL, L, SER, M, and MIH, or (s) MEL, SER, L and MIH. Control oocytes were incubated in the medium alone.