Fish sleeping under sandy bottom: interplay of melatonin and clock genes.

Wrasse species exhibit a definite daily rhythm in locomotor activity and bury themselves in the sand at the bottom of the ocean at night. It remains unclear how their behavior in locomotor activity is endogenously regulated. The aim of the present study was to clarify the involvement of melatonin and clock genes (Per1, Per2, Bmal1, and Cry1) in daily and circadian rhythms of the threespot wrasse, Halichoeres trimaculatus, which is a common species in coral reefs. Daily and circadian rhythms in locomotor activity were monitored under conditions of light-dark cycle (LD=12:12), constant light (LL), and darkness (DD). Daily rhythms in locomotor activity were observed under LD and persisted under LL and DD. Melatonin from a cultured pineal gland showed daily variations with an increase during the nighttime and a decrease during daytime, which persisted under DD. Melatonin treatment induced decreases in locomotor activity and respiratory rate, suggesting that melatonin has a sleep-inducing effect. Per1 and Per2 mRNA abundance in the brain under LD showed daily rhythms with an increase around lights on. Robust oscillation of Per1 and Per2 mRNA expression persisted under DD and LL, respectively. Expression of Bmal1 and Cry1 mRNA also showed daily and circadian patterns. These results suggest that clock genes are related to circadian rhythms in locomotor activity and that melatonin plays a role in inducing a sleep-like state after fish bury themselves in the sand. We conclude that the sleep-wake rhythm of the wrasse is regulated by a coordination of melatonin and clock genes.

Dopaminergic activity in the brain of a tropical wrasse in response to changes in light and hydrostatic pressure.

Many tropical wrasses show a daily pattern of spawning with gamete release typically near daytime high tide. The environmental cues the fish obtains from day-night and tidal cycles to ensure spawning synchrony and how those cues are transduced, however, are not fully understood. To gain insight into these issues, the involvement of monoamines in mediating endogenous day-night and tidal rhythms in the threespot wrasse, Halichoeres trimaculatus, were examined. Levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC, a metabolite of DA), serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA, a metabolite of 5-HT) in the brain of the fish were measured with high-performance liquid chromatography and electrochemical detection. DOPAC and the metabolic rate of DA activity (DOPAC/DA) were found to increase during the day and decrease during the night for fish held under a natural photoperiod. Fish acclimated to a 12:12 light-dark cycle and to constant dark conditions exhibited similar changes, whereas fish acclimated to constant light conditions exhibited little or no change. Intraperitoneal injection of melatonin resulted in a significant reduction in DOPAC/DA. Furthermore, DOPAC/DA was significantly lower in fish held at 3m compared to 0m depth, suggesting that hydrostatic pressure influences DA metabolic rate. These results indicate that light and hydrostatic pressure control dopaminergic turnover in the brain of threespot wrasse. Day-night and tidal changes in these two factors therefore may be the main environmental cues the fish uses to synchronize its spawning activity.

Role of the tidal cycle in the gonadal development and spawning of the tropical wrasse Halichoeres trimaculatus.

Many wrasses on coral reefs exhibit daily spawning that peaks around daytime high tides. In this study, we examined tidal-related ovarian development in the threespot wrasse, Halichoeres trimaculatus, a species common on coral reefs in the Indo-Pacific Ocean. When the fish were collected in the morning at different tidal phases, the gonadosomatic index (GSI) and ovarian histology changed; concomitant with increases in GSI towards high tide, a clutch of the most advanced oocytes developed from vitellogenic to maturation stages. Ovulated eggs and post-ovulated follicles (POF) existed in most ovaries around high tide, but only POF remained around ebb tide, suggesting that spawning occurred during or after high tide. We noticed that tidal-related spawning was considerable in the morning and that most ovaries collected on the afternoon high tide exhibited post-spawning features. This suggests that certain labrid species possess plasticity with regard to their spawning time and utilize potent environmental cues to ensure their reproductive success. When pieces of ovary were incubated with precursor steroids, high conversion of testosterone to 17beta-estradiol occurred during high and ebb tides, while that of 17alpha-hydroxyprogesterone to 17alpha,20beta-dihydroxy-4-pregnen-3-one and 17alpha,20beta,21-trihydroxy-4-pregnen-3-one was observed during low and flood tides. Incubation of pieces of ovary with human chorionic gonadotropin resulted in similar fluctuations in the steroid hormones with tidal phase. Production of these steroid hormones correlated with oocyte development in the ovaries and was probably regulated by gonadotropin. These results demonstrate that the daily cycle is fundamental for oocyte development, and that the tidal cycle is superimposed on this process.