Pituitary gonadotropin and ovarian gonadotropin receptor transcript levels: seasonal and photoperiod-induced changes in the reproductive physiology of female Atlantic salmon (Salmo salar).

In female Atlantic salmon kept at normal light conditions, pituitary follicle-stimulating hormone beta (fshb) transcript levels were transiently elevated one year before spawning, re-increased in February, and remained high during spawning in November and in post-ovulatory fish in December. The first increase in plasma 17b-estradiol (E2), testosterone (T) and gonadosomatic index (GSI) was recorded in January; E2 rose up to one month prior to ovulation, while T and GSI kept increasing until ovulation. Pituitary luteinizing hormone beta (lhb) transcript levels peaked at the time of ovulation. Except for transient changes before and after ovulation, ovarian follicle stimulating hormone receptor (fshr) transcript amounts were relatively stable at a high level. By contrast, luteinizing hormone receptor (lhcgr) transcript levels started out low and increased in parallel to GSI and plasma E2 levels. Exposure to continuous light (LL) induced a bimodal response where maturation was accelerated or arrested. The LL-arrested females showed previtellogenic oil droplet stage follicles or primary yolk follicles only, and fshb and E2 plasma levels collapsed while fshr increased. The LL-accelerated females showed elevated lhb transcript levels and slightly elevated E2 levels during early vitellogenesis, and significantly elevated lhcgr E2 and GSI levels in late vitellogenesis. We conclude that Fsh-dependent signaling stimulates recruitment into and the sustained development through vitellogenesis. Up-regulation of lhcgr gene expression during vitellogenesis may reflect an estrogenic effect, while elevated fshr gene expression following ovulation or during LL-induced arrestment may be associated with ovarian tissue remodeling processes.

Pituitary and plasma growth hormone dynamics during sexual maturation of female Atlantic salmon.

Growth hormone in fish regulates many important physiological processes including growth, metabolism and potentially reproduction. In salmonid fish, GH secretion is episodic with irregularly spaced GH peaks. Plasma GH reflects secretion episodes as well as the clearance rate of the hormone, and plasma levels may thus not always reflect the level of activation of the GH axis. This study measured the production dynamics of GH over a 17-month period in sexually maturing female Atlantic salmon which included final maturation and spawning. For the first time, the level of pituitary GH mRNA, pituitary GH protein and plasma GH protein were analyzed concurrently in the same individuals. mRNA and protein were extracted in parallel from the same samples with subsequent real time quantitative PCR to measure mRNA transcripts and radioimmunoassay to measure pituitary and plasma GH protein. Further, the effects of photoperiod manipulation on these parameters were studied. The results show no correlation between mRNA and protein levels except at some time points, and indicate that it is inappropriate to correlate pooled temporal data and averages in time series unless the relationship among the variables is stable over time. The results indicate complex and shifting relationships between pituitary GH mRNA expression, pituitary GH content and plasma GH levels, which could result from changes in clearance rather than secretion rate at different times and its episodic secretion. The study also suggests that there is a functionally important activation of the GH system during spring leading up to maturation and spawning.

Cloning of somatolactin alpha, beta forms and the somatolactin receptor in Atlantic salmon: seasonal expression profile in pituitary and ovary of maturing female broodstock.

BACKGROUND: Somatolactin (Sl) is a fish specific adenohypophyseal peptide hormone related to growth hormone (Gh). Some species, including salmonids, possess two forms: Sl alpha and Sl beta. The somatolactin receptor (slr) is closely related to the growth hormone receptor (ghr). Sl has been ascribed many physiological functions, including a role in sexual maturation. In order to clarify the role of Sl in the sexual maturation of female Atlantic salmon (Salmo salar), the full length cDNAs of slr, Sl alpha and Sl beta were cloned and their expression was studied throughout a seasonal reproductive cycle using real-time quantitative PCR (RTqPCR). METHODS: Atlantic salmon Sl alpha, Sl beta and slr cDNAs were cloned using a PCR approach. Gene expression of Sl alpha, SL beta and slr was studied using RTqPCR over a 17 month period encompassing pre-vitellogenesis, vitellogenesis, ovulation and post ovulation in salmon females. Histological examination of ovarian samples allowed for the classification according to the degree of follicle maturation into oil drop, primary, secondary or tertiary yolk stage. RESULTS: The mature peptide sequences of Sl alpha, Sl beta and slr are highly similar to previously cloned salmonid forms and contained the typical motifs. Phylogenetic analysis of Atlantic salmon Sl alpha and Sl beta shows that these peptides group into the two Sl clades present in some fish species. The Atlantic salmon slr grouped with salmonid slr amongst so-called type I ghr. An increase in pituitary Sl alpha and Sl beta transcripts before and during spawning, with a decrease post-ovulation, and a constant expression level of ovarian slr were observed. There was also a transient increase in Sl alpha and Sl beta in May prior to transfer from seawater to fresh water and ensuing fasting. CONCLUSION: The up-regulation of Sl alpha and Sl beta during vitellogenesis and spawning, with a subsequent decrease post-ovulation, supports a role for Sl during gonadal growth and spawning. Sl could also be involved in calcium/phosphate mobilization associated with vitellogenesis or have a role in energy homeostasis associated with lipolysis during fasting. The up-regulation of both Sl alpha and Sl beta prior to fasting and freshwater transfer, suggests a role for Sl linked to reproduction that may be independent of the maturation induced fasting.

A peak in gh-receptor expression is associated with growth activation in Atlantic salmon vertebrae, while upregulation of igf-I receptor expression is related to increased bone density.

Growth hormone (GH) and insulin-like growth factor-I (IGF-I) play major roles in the endocrine regulation of fish growth, but their interdependency and mode of action has not been well elucidated. The GH-IGF-I system is essential for normal vertebral growth in mouse, but this has not been studied in fish. To study the interplay between GH, IGF-I, and their receptors, postsmolt Atlantic salmon were studied during spring growth (January-June 2003). From January to June, fish were sampled regularly for plasma and vertebral bone. The vertebra was collected from the same anterior-posterior position. The growth hormone receptor (ghr) (There is no determined nomenclature of salmon genes but we stick to the nomenclature which is consequent for zebrafish, where all gene names are named with small letters and in italic.) expression in the vertebrae peaked in the end of February coinciding with high levels of plasma GH and IGF-I, and an increase of vertebral growth rate. From April to June, plasma IGF-I levels decreased together with ghr expression in the vertebrae, while plasma GH did not decrease. In May and June, expression of the igf-I receptor (igf-Ir) increased 4- to 5-fold, which coincided with an increase in bone density. The changes seen in gene expression of the IGF-I and GH receptors suggest that these hormones are involved in vertebral growth and bone density.