Increased expression of kisspeptin and GnRH forms in the brain of scombroid fish during final ovarian maturation and ovulation

BACKGROUND: Kisspeptins (Kiss) are prime players in the control of reproductive function through their regulation of gonadotropin-releasing hormone (GnRH) expression in the brain. The experimental scombroid fish, chub mackerel (Scomber japonicus) expresses two kiss (kiss1 and kiss2) and three gnrh (gnrh1, gnrh2, and gnrh3) forms in the brain. In the present study, we analyzed expression changes of kiss and gnrh mRNAs in the brain and corresponding GnRH peptides in the brain and pituitary during final ovarian maturation (FOM) and ovulation. METHODS: Female fish possessing late vitellogenic oocytes were injected with GnRH analogue to induce FOM and ovulation. Fish were observed for daily spawning activities and sampled one week post-injection at germinal vesicle migration (GVM), oocyte hydration, ovulation, and post-ovulatory time periods. Changes in relative mRNA levels of kiss and gnrh forms in the brain were determined using quantitative real-time PCR. Changes in GnRH peptides in the brain and pituitary were analyzed using time-resolved fluoroimmunoassay. RESULTS: Both kiss1 and kiss2 mRNA levels in the brain were low at late vitellogenic stage and increased significantly during the GVM period. However, kiss1 mRNA levels decreased during oocyte hydration before increasing again at ovulatory and post-ovulatory periods. In contrast, kiss2 mRNA levels decreased at ovulatory and post-ovulatory periods. Levels of gnrh1 mRNA in the brain increased only during post-ovulatory period. However, levels of gnrh2 and gnrh3 mRNAs were elevated during GVM and then, decreased during oocyte hydration before increasing again at ovulatory period. During post-ovulatory period, both gnrh2 and gnrh3 mRNA levels declined. Peptide levels of all three GnRH forms in the brain were elevated during GVM and oocyte hydration; their levels were significantly lower during late vitellogenic, ovulatory, and post-ovulatory periods. In contrast, pituitary GnRH peptide levels did not show any significant fluctuations, with the GnRH1 peptide levels being many-fold higher than the GnRH2 and GnRH3 forms. CONCLUSION: The results indicate increased expression of multiple Kiss and GnRH forms in the brain and suggest their possible involvement in the regulation of FOM and ovulation in captive female chub mackerel

Divulging the social sex change mechanism in a unique model system for studying the sexual plasticity of protogynous hermaphrodite fish, three bamboo leaf wrasse (Pseudolabrus sieboldi)

The gonadal sexual fate of vertebrates is either defined by genetics or environment, or a combination of both factors. Interestingly, in sequential hermaphroditism, the animal can undergo natural sex changes from female-to-male, male-to-female, and bidirectional way throughout their lives. This change exhibits the process which shifts between oogenesis and spermatogenesis and is regarded as an ideal instance of sexual plasticity. To develop the experimental model for studying the sexual plasticity of protogynous fish, the social conditions that induce sex changes were defined in wrasse, Pseudolabrus sieboldi. When six females were kept together in a tank, the largest female became a male, whereas a similar conversion did not occur when only two females were present in a tank. A semi-gonadectomy analysis developed in the present study verified the direct relationship between gonadal sex and body coloration. In P. sieboldi, the sex change is controlled by the relative body size of an individual within a group, rather than by absolute body size. When six females were kept in smaller sized tanks, delayed sex change or unchanged individuals was observed. Overall, more than 90% of the largest females demonstrated sex change after being housed with five smaller females in different sizes of tanks ranging from 80 to 500 L. Furthermore, the experiment using a transparent barrier suggested that visual stimuli are one of the major cues to initiate sex change. Our findings on the laboratory conditions leading to the initiation of sex change in wrasse suggest the usefulness of this species as a model organism for comparative studies in molecular, cellular, and physiological mechanisms of sexual plasticity, as well as on social and reproductive behaviors

The Roles of Kisspeptin System in the Reproductive Physiology of Fish With Special Reference to Chub Mackerel Studies as Main Axis

Kisspeptin, a novel neuropeptide product of the Kiss1 gene, activates the G protein-coupled membrane receptor G protein-coupled receptor 54 (now termed Kiss1r). Over the last 15 years, the importance of the kisspeptin system has been the subject of much debate in the mammalian research field. At the heart of the debate is whether kisspeptin is an absolute upstream regulator of gonadotropin-releasing hormone secretion, as it has been proposed to be the master molecule in reproductive events and plays a special role not only during puberty but also in adulthood. The teleostean kisspeptin system was first documented in 2004. Although there have been a number of kisspeptin studies in various fish species, the role of kisspeptin in reproduction remains a subject of controversy and has not been widely recognized. There is an extensive literature on the physiological and endocrinological bases of gametogenesis in fish, largely derived from studying small, model fish species, and reports on non-model species are limited. The reason for this discrepancy is the technical difficulty inherent in developing rigorous experimental systems in many farmed fish species. We have already established methods for the full life-cycle breeding of a commercially important marine fish, the chub mackerel (cm), and are interested in understanding the reproductive function of kisspeptins from various perspectives. Based on a series of experiments clarifying the role of the brain–pituitary–gonad axis in modulating reproduction in cm, we theorize that the kisspeptin system plays an important role in the reproduction of this scombroid species. In this review article, we provide an overview of kisspeptin studies in cm, which substantially aids in elucidating the role of kisspeptins in fish reproduction

Sex Lethal Gene Manipulates Gonadal Development of Medaka, Oryzias latipes, through Estrogenic Interventions

Germ cells are pivotal for gonadal sexuality maintenance and reproduction. Sex lethal (sxl), the somatic sex determining gene of Drosophila, is the known regulator and initiator of germ cell femininity in invertebrates. However, the role of the Sxl homologue has rarely been investigated in vertebrates. So, we used medaka to clarify the role of sxl in vertebrate gonadogenesis and sexuality and identified two Sxl homologues, i.e., Sxl1a and Sxl1b. We found that sxl1a specifically expresses in the primordial germ cells (PGC), ovary, (early gonia and oocytes), while sxl1b distributions are ubiquitous. An mRNA overexpression of sxl1a accelerated germ cell numbers in 10 DAH XY fish, and sxl1a knockdown (KD), on the other hand, induced PGC mis-migration, aberrant PGC structuring and ultimately caused significant germ cell reduction in XX fish. Using an in vitro promoter analysis and in vivo steroid treatment, we found a strong link between sxl1a and estrogenic germ cell-population maintenance. Further, using sxl1a-KD and erβ2-knockout fish, we determined that sxl1 acts through erβ2 and controls PGC sexuality. Cumulatively, our study highlights the novel role of sxl1a in germ cell maintenance and sexual identity assignment and thus might become a steppingstone to understanding the commonalities of animal sexual development

Analysis of the hatching site and migratory behaviour of the swordtip squid (<i>Uroteuthis edulis</i>) caught in the Japan Sea and Tsushima Strait in autumn estimated by statolith analysis

<p>In contrast to the swordtip squid (<i>Uroteuthis edulis</i>) caught in the spring and summer, those caught in the autumn in the Japan Sea and the Tsushima Strait are characterized by thick bodies with large clubs, long tentacles and large suckers. In addition, most females and many males are immature. However, the reasons for these characteristics, as well as the hatching site and migratory behaviour, are not well known. The empirical water temperature of juveniles was estimated at ∼17°C through strontium:calcium (Sr:Ca) ratios in statoliths and information on seawater temperature distribution. The ontogenetic variation in Sr:Ca ratios decreased from hatching to 60 days after hatching and then remained stable. The estimated water temperature was ∼21°C after 60 days. Thus, individuals of the autumn-migrating group most likely hatch in the southern East China Sea in the winter and early spring and then move north-eastwards with currents and pass through the Tsushima Strait into the southern Japan Sea during or after July. However, the squid could be present in these waters without strong currents northeast of Tsushima Island, where thermoclines occur during summer, and become confined in limited layers until vertical mixing occurs in the autumn. The distribution of the squid would thereafter extend to fishing grounds in the southern and south-eastern Japan Sea and then to the Tsushima Strait. This migratory behaviour could be the cause of the observed characteristics because bottom seawater temperatures are much lower in the Japan Sea than in the East China Sea.</p