Melatonin signal transduction in the goldfish, Carassius auratus.

Generation and reception of melatonin signals in the goldfish, Carassius auratus, are reviewed. The photoreceptive pineal gland of the goldfish generates circulating melatonin rhythms according to a given photoperiod under light-dark cycles and in a circadian manner under continuous dark conditions. Melatonin is also produced in the retina in a similar fashion. Melatonin produced in the pineal gland and retina is considered to act as internal zeitgeber in the brain and retina, respectively, controlling various physiological events via specific melatonin binding sites that are coupled with G protein. The goldfish exhibit clear diurnal locomotor activity rhythms under light-dark cycles and free-running rhythms under constant conditions. However, the relationship between melatonin and locomotor activity rhythms in the goldfish remains unclear. Further studies should be required to demonstrate the roles of melatonin in the circadian system in this species.

Melatonin secretion from goldfish pineal gland in organ culture.

Pineal glands were removed from goldfish reared under 12L-12D at 25 degrees for 2 weeks. These were incubated for 6 days under (1) normal 12L-12D (lights on 0600-1800 hr), (2) reversed 12L-12D (lights on 1800-0600 hr), (3) continuous dark, or (4) continuous light condition at 25 degrees. The incubation medium was changed at 12-hr intervals (0600-1800 and 1800-0600 hr) and secreted melatonin (MLT) was measured by RIA. Under 12L-12D or reversed 12L-12D, MLT secretion was active in the dark phase and was suppressed in the light phase of a given photoperiod. Under a continuous dark condition, a large amount of MLT was secreted into the medium, although the amount gradually decreased. The MLT secretion was more active in the period corresponding to the dark phase of the acclimatory photoperiod than in the period corresponding to the light phase. This pattern in secretion remained for 4 days. Under a continuous light condition, MLT secretion was suppressed, but the secretion was rapidly increased after changing the photoperiod from the light to the dark condition. These findings clearly indicate that MLT secretion in the organ-cultured pineal gland is photosensitive. It is active under dark and inactive under light conditions. The existence of a circadian rhythm in MLT secretion is also suggested.

Daily cycles in plasma melatonin levels under long or short photoperiod in the common carp, Cyprinus carpio.

A radioimmunoassay (RIA) for plasma melatonin (MLT) was simplified for use with the common carp (Cyprinus carpio). Plasma was partially purified with Sep-Pak C18 cartridge before RIA. The inhibition curves for the Sep-Pak C18 fraction from the plasma of carp, goldfish (Carassius auratus), yellow tail (Seriola quinqueradiata), Japanese eel (Anguilla japonica), bora (Mugil cephalus cephalus), kisu (Sillago japonica), ishigarei (Kareius bicoloratus), and shimaisaki (Rhyncopelates oxyrhynchus) were parallel with the MLT standard curve. There was a highly significant correlation between MLT added to carp plasma and that which was recovered (r = 0.997, P less than 0.01). Intraassay coefficients of variation at low, medium, and high levels were 4.7, 5.2, and 6.4, respectively. Interassay coefficients of variation at low, medium, and high levels were 11.8, 8.2, and 24.1%, respectively. The lower limit of detection was 11 pg/tube. Plasma MLT levels were investigated every 2 hr in carp under 16L-8D and 8L-16D at 24 degrees. Under both photoperiods, MLT levels showed marked daily patterns, i.e., the levels were high (220-540 pg/ml) during the dark phase and low (23-104 pg/ml) during the light phase. Tricaine methanesulfonate markedly interfered with the MLT measurements, while ethylcarbamate did not show any significant influence. Sexual difference in plasma MLT levels in carp was not observed. These observations suggest that MLT is an important hormone in photoperiodism and/or a circadian rhythm in fish.

Circadian rhythms of melatonin secretion from superfused goldfish (Carassius auratus) pineal glands in vitro.

A flow-through, whole-organ culture (superfusion) system was developed, and goldfish pineal glands were maintained at 25 degrees under light-dark (LD) 12:12 cycles, reversed LD 12:12 cycles, continuous dark (DD), or continuous light (LL) conditions for 48 hr. Under LD 12:12 and reversed LD 12:12 cycles, superfused pineal glands showed a rhythmic melatonin secretion: Scotophase was associated with high titers and photophase with low titers. The melatonin secretion rhythms persisted for two cycles under DD conditions, whereas nocturnal rises were suppressed under LL conditions. After the transition from LL to DD conditions on the third day, melatonin showed a nocturnal increase. These results indicate that melatonin secretion from the superfused goldfish pineal gland is directly photosensitive and that the goldfish pineal gland harbors a circadian oscillator which generates melatonin secretion rhythms.