Munehico Iwata

Circadian Changes in Serum Concentrations of Steroids in Japanese Char Salvelinus leucomaenis at the Stage of Final Maturation

Abstract Circadian changes in serum concentrations of testosterone (T), 11-ketotestosterone (11KT), estradiol-17β (E2), 17α,20β-dihydroxy-4-pregnen-3-one (DHP), 17α-hydroxyprogesterone (OHP), cortisol (F) and progesterone (P) were investigated in the spermiated/ovulated Japanese char Salvelinus leucomaenis for over three days using newly developed time-resolved fluoroimmunoassays. Testosterone and DHP in both sex and 11KT in male showed significantly (P<0.05) higher serum levels just before/after onset of darkness (15:00 or 18:00), and the levels during night and daytime were significantly (P<0.05) lower than those of the peak levels. Serum F levels in both sex during dark phase were significantly (P<0.05) higher than those levels during daytime. A surge of serum OHP concentrations in both sexes was observed at the time of twilight (03:00). The peak time of serum T, 11KT and DHP levels were approximately 6 hours prior to those of serum F and OHP levels. Serum E2 in female and P in both sex fluctuated intensely during sampling period, and did not show remarkable changes. These results strongly suggest the existence of circadian-like diel changes in serum T, DHP, F and OHP levels in both sex and 11KT in male, and no variations in serum E2 in female and P in both sex in spermiated/ovulated Japanese char under the stage of final maturation. Furthermore, relationship between circadian rhythms of steroid hormones and spawning behaviors are discussed in the present study.

Gonadotropin-releasing hormone (GnRH): expression during salmon migration

In the seaward migrating chum salmon, immunocytochemical and in situ hybridization techniques revealed isolated GnRH neurons at the base of the nasal epithelium, along the nervus terminalis and as ganglia at the rostroventral (gROB) and caudalmost (GT) olfactory bulb. A novel GnRH ganglion was seen at the cribriform bone (gCB). GnRH immunoreactivity but not the hybridization signal was detected in the midbrain neurons. During the migratory period, there were trends towards an increase in GnRH mRNA in the gCB and the gROB and a significant surge in plasma thyroid hormones was also evident. Therefore, we hypothesise thyroid hormones might be crucial for the increased tendency of GnRH expression and the migratory behavior of chum salmon.

Effect of thyroxine on the decrease of aggressive behaviour of four salmonids during the parr-smolt transformation

Abstract The effect of thyroid hormone on the aggressive behaviour of four salmonid species from three genera was investigated during the pre-migratory period. One-year-old pre-smolts of sea-run brown trout ( Salmo trutta ), steelhead trout ( Oncorhynchus mykiss ), and masu salmon ( Onchorynkus masou ), and one-year-old non-anadromous brook charr ( Salvelinus fontinalis ) were administered thyroxine (T 4 ;1 mg/g food) over three days. T 4 -treated brown trout, steelhead trout and masu salmon showed a significant reduction in aggressive behaviour compared to control fish. Non-anadromous brook charr showed no change in aggression after T 4 treatment. These data suggest that plasma T 4 elevation during smolting reduces aggressive behaviour and downstream migration, but T 4 does not affect the aggressive behaviour of non-anadromous brook charr.

Influence of modern irrigation, drainage system and water management on spawning migration of mud loach, Misgurnus anguillicaudatus C.

We investigated the influence of human activities on the spawning migration of the mud loach, Misgurnus anguillicaudatus. Mud loaches migrate from rivers to paddy fields for spawning. In this study, the number of adult loaches in paddy fields showed a peak in late May and rapidly decreased in July. In early July, water is discharged from paddy fields in order to allow the fields to dry for a few weeks, and water is again supplied later. The spawning season of the loaches probably spans from mid May to August; this was concluded by observing physiological indicators such as the gonad somatic index, oocyte maturity and serum levels of testosterone, estradiol-17β and 17α, 20β-dihydroxy-4-pregnen-3-one. When the loaches mature, they migrated to paddy fields if they could swim upward through the drainage ditches. However, the drainage ditches of modern paddy fields were reconstructed to deepen the water levels. The artificial drought of the rice culture management led to a discharge of water and loaches from the paddy fields during their spawning season. These results indicate that modern rice cultivation causes obstruction of the migration route and loss of spawning grounds; consequently, loaches use limited space and period of the paddy field for their spawning.

Function and trigger of thyroxine surge in migrating chum salmon Oncorhynchus keta fry

Abstract Chum salmon, Oncorhynchus keta, fry were reared in the Otsuchi Hatchery, fed for 5 weeks after yolk absorption, and then released at 09:30 to a hatchery pond. Diel rhythm of plasma thyroxine (T4) was not observed in fresh water or in seawater. Plasma T4 levels were less than 3 ng/ml in fish reared in well water in the hatchery raceway, whereas the level was in a range of 2–6 ng/ml and showed a T4 surge of 14.3±1.4 ng/ml when they were reared in stream water. Fry released from the raceway began to form schools, and then actively swam downstream headfirst. The T4 levels of early migrants increased from 2–7 ng/ml to 20–35 ng/ml during downstream migration in the afternoon in three study years. In 2001, we sampled two groups of fry that spent 2 h for school formation in the hatchery pond, and a T4 surge was observed. In both cases, the T4 levels decreased in the late afternoon during downstream migration to the river mouth. Plasma T4 increased again in the Fisherman Port before departure to the Otsuchi Bay. Another T4 increase was detected in the bay before offshore emigration. Stimuli such as rainfall, turbid water and cold water also triggered T4 surges. The duration of the T4 surge was short, usually less than two h. These results suggest that T4 surges occur when fry schools change their habitats.

INTRACEREBRAL EXPRESSION OF GONADOTROPIN-RELEASING HORMONE AND GROWTH HORMONE-RELEASING HORMONE IS DELAYED UNTIL SMOLTIFICATION IN THE SALMON

A developmental strategy was employed to investigate the functional assembly of neuropeptidergic systems in the migratory species of chum salmon Oncorhynchus keta. Using immunocytochemistry we have demonstrated that different groups of gonadotropin-releasing hormone- (GnRH)- and growth hormone-releasing hormone- (GHRH)-synthesizing neurons emerged according to very different developmental timetables. From the eye pigmentation stage (23 +/- 2 days after fertilisation (DAF)) through to the pre-smoltification stage (136 DAF), salmon-GnRH neurons originating from the olfactory placodes remained restricted to the extracerebral course of the terminal nerve. At the climax of smoltification (downstream migration 167 DAF), basal forebrain and midbrain GnRH neurons with elaborate neurite outgrowths in the brain and the pituitary became detectable. The GnRH neuroanatomical organization in the post-smoltification stage (197 DAF) was similar to that in the smoltification stage (167 DAF). In contrast to the case for other teleosts, chicken-GnRHII neurons were not found in the midbrain but were localized along the medial regions of the olfactory nerve. Growth hormone-releasing hormone immunoreactivity in the olfactory apparatus (21 DAF), and fibers along the basal telencephalon and hypothalamus and in the pituitary were observed during early embryogenesis (51 DAF) and in cells in the preoptic area on 167 DAF. The intracerebral expression of GnRH and GHRH was not detected until the peak of smoltification, which coincided with a peak in thyroid hormones, and precisely with downstream migratory behavior.

A comparative analysis of aggression in migratory and non-migratory salmonids

This paper represents a comprehensive test of the hypothesis that aggression in salmonids increases with the duration of stream residence. The intraspecific aggression of eleven juvenile salmonids was compared with their normal duration of stream residence. Salmonids maintained in 1 metre tanks and observed over two days could be separated into four groups based on statistical differences between the frequencies of aggressive behaviour. Non-anadromous Salvelinus fontinalis showed the highest levels of aggressive behaviour, followed by Oncorhynchus masou and O. mykiss (rainbow and steelhead trout). Least aggressive were the early migrants O. nerka (kokanee and sockeye salmon) and O. keta. The remaining species, O. rhodurus, O. kisutch, S. leucomaenis pluvius and Salmo trutta formed a moderately aggressive group. Aggressiveness was significantly and positively correlated with the duration of stream residence.

Lack of Circadian Regulation of Melatonin Rhythms in the Sockeye Salmon (Oncorhynchus nerka) in vivo and in vitro

Abstract Melatonin profiles were determined in the plasma in vivo and in the pineal organ in vitro of the sockeye salmon (Oncorhynchus nerka) under various light conditions to test whether they are under circadian regulation. When serial blood samples were taken at 4-h intervals for 3 days via a cannula inserted into the dorsal aorta, plasma melatonin exhibited significant fluctuation under a light-dark cycle, with higher levels during the dark phase than during the light phase. No rhythmic fluctuations persisted under either constant dark or constant light, with constant low and high levels, respectively. Melatonin release from the pineal organ in flow-through culture exhibited a similar pattern in response to the change in light conditions, with high and low release associated with the dark and light phases, respectively. These results indicate that melatonin production in the sockeye salmon is driven by light and darkness but lacks circadian regulation.

Central administration of growth hormone-releasing hormone and corticotropin-releasing hormone stimulate downstream movement and thyroxine secretion in fall-smolting coho salmon (Oncorhynchus kisutch)

The ultimate signal triggering downstream migration in anadromous salmonids is unknown. A plasma surge of T(4) (T(4) surge) occurs during downstream migration in salmonids; however, the causal relationship between migratory behavior and the T(4) surge is not well known. We first examined the progression of smolt indicators (skin silvering, condition factor (CF), gill Na(+), K(+)-ATPase (NKA) activity and plasma T(4) levels) in underyearling, fall-smolting coho salmon (Oncorhynchus kisutch) from August to December. In November, the fish showed the characteristics of fully developed smolts, i.e. the skin completely covered with silvery scales, CF at a nadir, and peak NKA activity and plasma T(4) levels. Based on these results, we examined the effects of four neuropeptides, thyrotropin-releasing hormone (TRH), corticotropin-releasing hormone (CRH), growth hormone-releasing hormone (GHRH), and gonadotropin-releasing hormone (GnRH), on the downstream movement (negative rheotaxis) and T(4) surge in fully smoltified underyearling coho salmon. The experiment was run in circular-shaped channel tanks and the neuropeptide treatment was performed as intracerebroventricular (ICV) injections. ICV injection of GHRH and CRH stimulated both downstream movement and plasma T(4) level. TRH injection stimulated plasma T(4) level but suppressed downstream movement. GnRH injection had no effect. It is hypothesized that GHRH and CRH play key roles in triggering downstream migration of anadromous salmonids, and that the accompanying T(4) surge is a consequence of the neuroendocrine processes that trigger migration.

Central administration of growth hormone-releasing hormone triggers downstream movement and schooling behavior of chum salmon (Oncorhynchus keta) fry in an artificial stream.

Anadromous salmonids migrate downstream to the ocean (downstream migration). The neuroendocrine mechanism of triggering the onset of downstream migration is not well known. We investigated the effects of 14 chemicals, including neuropeptides, pineal hormones, neurotransmitters, and neuromodulators (growth hormone-releasing hormone: GHRH, thyrotropin-releasing hormone, corticotropin-releasing hormone: CRH, gonadotropin-releasing hormone, melatonin, N-acetyl serotonin, serotonin, beta-endorphin, enkephalin, dopamine, norepinephrine, epinephrine, acetylcholine, and histamine) on the onset of downstream migration in chum salmon (Oncorhynchus keta) fry. We defined downstream migration as a downstream movement (negative rheotaxis) with schooling behavior and counted the number of downstream movements and school size in experimental circulation tanks. An intracerebroventricular injection of GHRH, CRH, melatonin, N-acetyl serotonin, or serotonin stimulated the number of downstream movements. However, GHRH was the only chemical that also stimulated an increase in schooling behavior. These results suggest that CRH, melatonin, N-acetyl serotonin, and serotonin are involved in the stimulation of downstream movement in chum salmon, while GHRH stimulates both downstream movement and schooling behavior.