Effects of fasting on the central expression of appetite-regulating and reproductive hormones in wild-type and Casper zebrafish (Danio rerio)

Abstract

Appetite and reproduction are closely related functions that are both regulated by brain hormones. Appetite stimulators include orexin and neuropeptide Y (NPY), and reproductive hormones include gonadotropin-releasing hormone (GnRH), gonadotropin-inhibitory hormone (GnIH), kisspeptin, and neurokinin B (NKB). GnRH stimulates the secretion of pituitary gonadotropes, and kisspeptin and GnIH modulate this action. Kisspeptin secretion is further controlled by neurokinin B (NKB) and dynorphin A (Dyn). To better understand the mechanisms regulating appetite and reproduction in fish, we examined the effects of fasting, reproductive stage, gender, and strain on the brain mRNA expression of appetite (orexin and NPY) and reproductive (GnRH, kisspeptin, GnIH, and NKB) hormones in zebrafish. In order to compare strains, we used both wild-type and transparent Casper zebrafish. In female wild-type zebrafish, fasting increased the expression of all hormones investigated, with the exception of Kiss2. Only NPY and Kiss2 were increased in male wild-type zebrafish during fasting. In Casper zebrafish, only GnIH and NKB in males were affected by fasting, suggesting that Casper fish may be more resistant to fasting than wild fish. Fasting increased expressions of orexin, GnRH2, Kiss1, GnIH and NKB in wild-type females with more eggs or larger eggs relative to body weight, compared to those with fewer or smaller eggs, suggesting that more mature females are more affected by fasting. No significant interactions of fasting and reproductive stage were noted in female Casper fish. To investigate whether differences between Casper and wild-type fish were due to genes involved in pigmentation, we compared the brain mRNA expressions of enzymes involved in melanin synthesis (tyrosinase and tyrosine hydroxylase - TH), melanocortin receptors (MC3R and MC4R), and the melanocortin precursor (proopiomelanocortin - POMC) between the two strains. Casper zebrafish had lower levels of MC3R, tyrosinase, TH1, TH2, and POMC than wild-type fish. Overall, our results suggest the existence of gender- and reproductive stage-specific, as well as strain-specific variations in the mechanisms regulating feeding and reproduction in zebrafish, and that the melanocortin system and melanin pathways may be in part responsible for these differences between strains.

Keywords: Appetite; Endocrine regulation; Melanocortin; Reproduction; Zebrafish; mRNA expression.