Effect of intracerebroventricular administration of two molecular forms of sulfated CCK octapeptide on anxiety-like behavior in the zebrafish danio rerio.

Cholecystokinin octapeptide with sulfate (CCK-8s) regulates feeding behavior and psychomotor activity. In rodents and goldfish, intracerebroventricular (ICV) injection of CCK-8s decreases food intake and also induces anxiety-like behavior. The zebrafish has several merits for investigating the psychophysiological roles of neuropeptides. However, little is known about the brain localization of CCK and the behavioral action of CCK-8s in this species. Here we investigated the brain localization of CCK-like immunoreactivity and found that it was distributed throughout the brain. As CCK-like immunoreactivity was particularly evident in the ventral habenular nucleus, the interpeduncular nucleus and superior raphe, we subsequently examined the effect of zebrafish (zf) CCK-8s on psychomotor control. Since the zebrafish possesses two molecular forms of zfCCK-8s (zfCCKA-8s and zfCCKB-8s), two synthetic peptides were administered intracerebroventricularly at 1, 5 and 10 pmol g-1 body weight (BW). As the zebrafish shows a greater preference for the lower area of a tank than for to the upper area, we used this preference for assessment of anxiety-like behavior. ICV administration of zfCCKA-8 s or zfCCKB-8s at 10 pmol g-1 BW significantly shortened the time spent in the upper area. The actions of these peptides mimicked that of the central-type benzodiazepine receptor inverse agonist FG-7142 (an anxiogenic agent) at 10 pmol g-1 BW. The anxiogenic-like action of the two peptides was attenuated by treatment with the CCK receptor antagonist proglumide at 200 pmol g-1 BW. These results indicate that zfCCKA-8s and zfCCKB-8s potently induce anxiety-like behavior via the CCK receptor-signaling pathway in the zebrafish brain.

Neuroendocrine control of feeding behavior and psychomotor activity by neuropeptideY in fish.

Neuropeptide Y (NPY) is a neuropeptide distributed widely among vertebrates. In mammals, NPY and its related peptides such as pancreatic polypeptide and peptide YY (PYY) are distributed throughout the brain and gastrointestinal tissues, and are centrally involved in many physiological functions such as the regulation of food intake, locomotion and psychomotor activities through their receptors. With regard to non-mammalian vertebrates, there has also been intensive study aimed at the identification and functional characterization of NPY, PYY and their receptors, and recent investigations of the role of NPY have revealed that it exerts several behavioral effects in goldfish and zebrafish. Both of these species are excellent teleost fish models, in which it has been demonstrated that NPY increases food consumption as an orexigenic factor and reduces locomotor activity, as is the case in mammals. This paper reviews current knowledge of NPY derived from studies of teleost fish, as representative non-mammals, focusing particularly on the role of the NPY system, and examines its significance from a comparative viewpoint.

Gonadotropin-releasing hormone 2 suppresses food intake in the zebrafish, Danio rerio.

Gonadotropin-releasing hormone (GnRH) is an evolutionarily conserved neuropeptide with 10 amino acid residues, of which several structural variants exist. A molecular form known as GnRH2 ([His(5) Trp(7) Tyr(8)]GnRH, also known as chicken GnRH II) is widely distributed in vertebrates except for rodents, and has recently been implicated in the regulation of feeding behavior in goldfish. However, the influence of GnRH2 on feeding behavior in other fish has not yet been studied. In the present study, therefore, we investigated the role of GnRH2 in the regulation of feeding behavior in a zebrafish model, and examined its involvement in food intake after intracerebroventricular (ICV) administration. ICV injection of GnRH2 at 0.1 and 1 pmol/g body weight (BW) induced a marked decrease of food consumption in a dose-dependent manner during 30 min after feeding. Cumulative food intake was significantly decreased by ICV injection of GnRH2 at 1 pmol/g BW during the 30-min post-treatment observation period. The anorexigenic action of GnRH2 was completely blocked by treatment with the GnRH type I receptor antagonist Antide at 25 pmol/g BW. We also examined the effect of feeding condition on the expression level of the GnRH2 transcript in the hypothalamus. Levels of GnRH2 mRNA obtained from fish that had been provided excess food for 7 days were higher than those in fish that had been fed normally. These results suggest that, in zebrafish, GnRH2 acts as an anorexigenic factor, as is the case in goldfish.

Stimulatory effect of intracerebroventricular administration of orexin A on food intake in the zebrafish, Danio rerio.

Orexin is a potent orexigenic neuropeptide implicated in feeding regulation of mammals. However, except for the case of goldfish, the involvement of orexin in the feeding behavior of teleost fish has not well been studied. Therefore, we investigated the role of orexin on food intake using a zebrafish (Danio rerio) model. We examined the effect of feeding status on orexin-like immunoreactivity and the expression level of orexin transcript in the brain. The number of neuronal cells showing orexin-like immunoreactivity in the hypothalamic region, including the posterior tuberal nucleus, was significantly increased in fish fasted for 7days. Orexin precursor mRNA levels in the brain obtained from fish fasted for 7 days were higher than those in fish that had been fed normally. We then investigated the effect of intracerebroventricular (ICV) administration of orexin A on food intake. Cumulative food intake was significantly increased by ICV administration of orexin A (at 0.3 and 3 pmol/g body weight, BW) during a 60-min observation period after treatment. The orexin A-induced orexigenic action (at 0.3 pmol/g BW) was blocked by treatment with an orexin receptor antagonist, SB334867, at 10 pmol/g BW. These results indicate that orexin A acts as feeding regulator in the zebrafish.