gonadectomy-blood-sampling-procedures-small-size-teleost-model

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka (<em>Oryzias latipes</em>)

Sex steroids, produced by the gonads, play an essential role in brain and pituitary tissue plasticity and in the neuroendocrine control of reproduction in ...

University of Macau

Sex determination and differentiation are complex processes. As a juvenile hermaphrodite or undifferentiated gonochorist, zebrafish undergo a special juvenile ovarian phase during sex differentiation, making it an excellent model for studying early oogenesis and folliculogenesis. We provide lines of evidence at morphological, molecular, and genetic levels for roles of factor in the germline α (Figla), an oocyte-specific transcription factor, in early zebrafish gonadogenesis. As in mammals, Figla/figla was also expressed in the gonads and its expression in the ovary was also restricted to early oocytes. Disruption of figla gene by CRISPR/Cas9 led to an all-male phenotype in the mutant. Detailed analysis of early gonadal development showed that the germ cells in the mutant were clustered in cysts and underwent meiosis, forming oocytes at prefollicular chromatin nucleolar (CN) stage (stage IA). However, the subsequent transition from cystic CN oocytes to individual follicular perinucleolar oocytes (stage IB) was blocked, resulting in an all-male phenotype in the mutant. The phenotype of figla mutant could not be rescued by estrogen treatment, in contrast to cyp19a1a mutant, and introduction of tp53 mutation also had no effect, unlike in fancd1 and fancl mutants. Transcriptome analysis revealed that many biological processes and pathways related to germ cell development, especially oogenesis, were upregulated in the presence of Figla and that the regulation of figla expression may involve heat shock proteins. Our results strongly suggest important roles for Figla in juvenile ovary development, especially the formation of individual follicles from cystic oocytes.

Roles of Figla/figla in Juvenile Ovary Development and Follicle Formation During Zebrafish Gonadogenesis.

Bisphenol A (BPA) can induce endocrine disorders in humans and animals. In this study, we used several zebrafish mutants deficient in estrogen production and signalling, which could be valuable for evaluating estrogenic activities and mechanisms of EDCs. With low endogenous estrogens, the all-male aromatase mutant (cyp19a1a) is expected to be more responsive to estrogenic exposure, and mutants of nuclear estrogen receptors (nERs; esr1, esr2a and esr2b) alone or in combination would allow us to evaluate the action mechanisms of estrogenic EDCs. Exposure to BPA could rescue the all-male phenotype of the cyp19a1a mutant, delayed gonadal development in both sexes, resulting in infertility or subfertility, and caused follicle atresia in females and impairment of spermatogenesis in males. To understand the mechanisms of these effects, we tested BPA in cyp19a1a and nER mutants of different combinations. The feminizing effect of BPA on sexual differentiation was dependent on nERs, in particular esr2a. As for males, nERs were also involved in BPA-induced impairment of spermatogenesis. Taken together, with genome editing technology our study provides the most comprehensive genetic evidence for estrogenic activities of BPA in zebrafish and its action mechanisms. This study also establishes a powerful platform for studying other EDCs with estrogenic activity.

Genetic evidence for estrogenicity of bisphenol A in zebrafish gonadal differentiation and its signalling mechanism.

Sex determination and differentiation are complex processes controlled by many different factors; however, the relationships among these factors are poorly understood. Zebrafish gonadal differentiation exhibits high plasticity involving multiple factors and pathways, which provides an excellent model for investigating the interactions between them. Ovarian aromatase () and  are key factors in directing vertebrate ovary and testis differentiation, respectively. Knockout of zebrafish  leads to all-male offspring, whereas the loss of  results in a female-biased sex ratio. In the present study, we established  ; double mutant zebrafish and discovered that the introduction of the  mutation into the  mutant could rescue the all-male phenotype of the latter. Interestingly, despite the lack of aromatase/estrogens, the follicles in the ovary of the rescued  mutant could develop normally up to the previtellogenic stage. Further evidence suggested the ovarian aromatase directed ovarian differentiation by suppressing  expression via nuclear estrogen receptors (nERs). Our results provide solid evidence for an interaction between  and  in zebrafish gonadal differentiation, and for the dispensability of estrogens in controlling early folliculogenesis.

Disruption of  rescues the all-male phenotype of the  mutant in zebrafish  a novel insight into the roles of aromatase/estrogens in gonadal differentiation and early folliculogenesis.

Weiyi Song

Centre of Reproduction,
Development and Aging (CRDA),
Faculty of Health Sciences

University of Macau

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka (Oryzias latipes)

Weiyi Song

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Centre of Reproduction,Development and Aging (CRDA),Faculty of Health Sciences

University of Macau

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka (Oryzias latipes)

Centre of Reproduction,
Development and Aging (CRDA),
Faculty of Health Sciences