nr0b1 (DAX1) mutation in zebrafish causes female-to-male sex reversal through abnormal gonadal proliferation and differentiation

Chen, Sijie; Zhang, Hefei; Wang, Fenghua; Zhang, Wei; Peng, Gang

doi:10.1016/j.mce.2016.06.005

Cited by 40 publications

(23 citation statements)

References 81 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Disruption of zebrafish fancl and fancd1 (brca2) gene functions caused female-to-male sex reversal by p53-mediated germ cell apoptosis (Rodríguez-Marí et al 2010. However, nr0b1 (DAX1) mutation in zebrafish resulted in female-to-male sex reversal in a p53-independent manner (Chen et al 2016). Disruption of foxl2a and foxl2b function led to the sex reversal from females to males (Yang et al 2017).…”

Section: Discussionmentioning

confidence: 99%

Distinct and Cooperative Roles of amh and dmrt1 in Self-Renewal and Differentiation of Male Germ Cells in Zebrafish

et al. 2017

View full text Add to dashboard Cite

Spermatogenesis is a fundamental process in male reproductive biology and depends on precise balance between self-renewal and differentiation of male germ cells. However, the regulative factors for controlling the balance are poorly understood. In this study, we examined the roles of and in male germ cell development by generating their mutants with Crispr/Cas9 technology in zebrafish. mutant zebrafish displayed a female-biased sex ratio, and both male and female mutants developed hypertrophic gonads due to uncontrolled proliferation and impaired differentiation of germ cells. A large number of proliferating spermatogonium-like cells were observed within testicular lobules of the -mutated testes, and they were demonstrated to be both Vasa- and PH3-positive. Moreover, the average number of Sycp3- and Vasa-positive cells in the mutants was significantly lower than in wild-type testes, suggesting a severely impaired differentiation of male germ cells. Conversely, all the -mutated testes displayed severe testicular developmental defects and gradual loss of all Vasa-positive germ cells by inhibiting their self-renewal and inducing apoptosis. In addition, several germ cell and Sertoli cell marker genes were significantly downregulated, whereas a prominent increase of Insl3-positive Leydig cells was revealed by immunohistochemical analysis in the disorganized-mutated testes. Our data suggest that might act as a guardian to control the balance between proliferation and differentiation of male germ cells, whereas might be required for the maintenance, self-renewal, and differentiation of male germ cells. Significantly, this study unravels novel functions of gene in fish.

show abstract

Section: Discussionmentioning

confidence: 99%

Distinct and Cooperative Roles of amh and dmrt1 in Self-Renewal and Differentiation of Male Germ Cells in Zebrafish

et al. 2017

View full text Add to dashboard Cite

show abstract

“…Down- and up-regulation of cyp11a expression by cAMP administration indicates that flounder cyp11a expression could also be regulated by cAMP and the regulation was dosage dependent. NR5a2 (i.e., SF-1) and NR0b1 (i.e., Dax-1) are the mainly regulated factors in the steroidogenesis [ 34 ]. These two trans-regulators could regulate cyp11a transcriptional activities through their specific cis-elements or protein–protein interactions [ 11 ].…”

Section: Discussionmentioning

confidence: 99%

“…NR5a2 plays an important role in the expression of tissue-specific genes and regulates the expression of steroidogenic genes, such as cyp11a [ 11 ]. It is mainly expressed in the testis, ovary and adrenal cortex, controlling the tissue-specific transcription of cyp11a in these tissues [ 34 ]. The binding site (5′-CAAGGCC-3′) of NR5a2 was found in the promoter of flounder cyp11a by analyzing its genome sequence [ 19 ].…”

Section: Discussionmentioning

confidence: 99%

Characteristics of Cyp11a during Gonad Differentiation of the Olive Flounder Paralichthys olivaceus

Liang

Fan

Zou

et al. 2018

IJMS

View full text Add to dashboard Cite

The P450 side-chain cleavage enzyme, P450scc (Cyp11a) catalyzes the first enzymatic step for the synthesis of all steroid hormones in fish. To study its roles in gonads of the olive flounder Paralichthys olivaceus, an important maricultured fish species, we isolated the cyp11a genomic DNA sequence of 1396 bp, which consists of 5 exons and 4 introns. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) results indicated that the flounder cyp11a was exclusively expressed in gonad and head kidney tissues. Its expression level in the testis was higher than that in the ovary. According to the in situ hybridization patterns, cyp11a was mainly expressed in the Leydig cells of the testis, and the thecal cells of the ovary. Immunofluorescence analysis showed that Cyp11a was located in the cytoplasm of the cultured flounder testis cells. Further quantitative real-time PCR results presented the cyp11a differential expression patterns during gonad differentiation. Among different sampling points of the 17β-estradiol (E2, 5 ppm) treatment group, cyp11a expression levels were relatively high in the differentiating ovary (30 and 40 mm total length, TL), and then significantly decreased in the differentiated ovary (80, 100 and 120 mm TL, p < 0.05). The pregnenolone level also dropped in the differentiated ovary. In the high temperature treatment group (HT group, 28 ± 0.5 °C), the cyp11a expression level fluctuated remarkably in the differentiating testis (60 mm TL), and then decreased in the differentiated testis (80, 100 mm TL, p < 0.05). In the testosterone (T, 5 ppm) treatment group, the cyp11a was expressed highly in undifferentiated gonads and the differentiating testis, and then dropped in the differentiated testis. Moreover, the levels of cholesterol and pregnenolone of the differentiating testis in the HT and T groups increased. The expression level of cyp11a was significantly down-regulated after the cultured flounder testis cells were treated with 75 and 150 μM cyclic adenosine monophosphate (cAMP), respectively (p < 0.05), and significantly up-regulated after treatment with 300 μM cAMP (p < 0.05). Both nuclear receptors NR5a2 and NR0b1 could significantly up-regulate the cyp11a gene expression in a dosage dependent way in the testis cells detected by cell transfection analysis (p < 0.05). The above data provides evidence that cyp11a would be involved in the flounder gonad differentiation and development.

show abstract

“…A similar knockout strategy was also used in irf6 (Li et al, 2017 ) and hemogen (Peters et al, 2018 ) knockout zebrafish, which showed transcripts destabilization and degradation of truncated Irf6 and Hemogen by NMD, and significantly decreased protein production. Therefore, if a few truncated transcripts were translated, the proteins might have been rapidly degraded (Chen et al, 2016 ; Facchinello et al, 2017 ). However, it cannot exclude a weak dominant-negative effect on other target genes, as Sox18 missing part of the transactivation domain may act in a dominant-negative manner (Pennisi et al, 2000a ; Pennisi et al, 2000b ).…”

Section: Discussionmentioning

confidence: 99%

“…Domesticated zebrafish strains lack sex-linked loci, although natural strains have WZ/ZZ sex chromosomes (Wilson et al, 2014 ). A number of genes involved in gonadal differentiation have been identified in zebrafish, which include female sex-biased genes: mettl3 (methyltransferase like 3) (Xia et al, 2018 ), cyp19a1a (cytochrome P450, family 19, subfamily A, polypeptide 1a) (Lau et al, 2016 ; Yin et al, 2017 ), nr0b1 (nuclear receptor subfamily 0 group B member 1) (Chen et al, 2016 ), foxl2a (forkhead box L2a), foxl2b (forkhead box L2b) (Yang et al, 2017 ), bmp15 (bone morphogenic protein 15) (Dranow et al, 2016 ) and fgf24 (fibroblast growth factor 24) (Leerberg et al, 2017 ), and male sex-biased genes: dmrt1 (doublesex and mab-3 related transcription factor 1) (Guo et al, 2005 ; Lin et al, 2017 ; Webster et al, 2017 ), amh (anti-Mullerian hormone) (Lin et al, 2017 ), sox9a (sex-determining region Y-box 9a) (Sun et al, 2013 ) and ar (androgen receptor) (Crowder et al, 2018 ). As lack of morphological sex chromosome in zebrafish, molecular mechanisms of sex determination and differentiation are probably multigenic (Liew et al, 2012 ) and key genes remain to be identified.…”

Section: Introductionmentioning

confidence: 99%

Loss-of-function of sox3 causes follicle development retardation and reduces fecundity in zebrafish

Liu

Ying

et al. 2018

Protein Cell

View full text Add to dashboard Cite

Folliculogenesis is essential for production of female gametes in vertebrates. However, the molecular mechanisms underlying follicle development, particularly apoptosis regulation in ovary, remain elusive. Here, we generated sox3 knockout zebrafish lines using CRISPR/Cas9. sox3 knockout led to follicle development retardation and a reduced fecundity in females. Comparative analysis of transcriptome between sox3 −/− and wild-type ovaries revealed that Sox3 was involved in pathways of ovarian steroidogenesis and apoptosis. Knockout of sox3 promoted follicle apoptosis and obvious apoptosis signals were detected in somatic cells of stages III and IV follicles of sox3 −/− ovaries. Moreover, Sox3 can bind to and activate the promoter of cyp19a1a . Up-regulation of Cyp19a1a expression promoted 17β-estradiol synthesis, which inhibited apoptosis in follicle development. Thus, Sox3 functions as a regulator of Cyp19a1a expression, via 17β-E2 linking apoptosis suppression, which is implicated in improving female fecundity. Electronic supplementary material The online version of this article (10.1007/s13238-018-0603-y) contains supplementary material, which is available to authorized users.

show abstract

nr0b1 (DAX1) mutation in zebrafish causes female-to-male sex reversal through abnormal gonadal proliferation and differentiation

Cited by 40 publications

References 81 publications

Distinct and Cooperative Roles of amh and dmrt1 in Self-Renewal and Differentiation of Male Germ Cells in Zebrafish

Distinct and Cooperative Roles of amh and dmrt1 in Self-Renewal and Differentiation of Male Germ Cells in Zebrafish

Characteristics of Cyp11a during Gonad Differentiation of the Olive Flounder Paralichthys olivaceus

Loss-of-function of sox3 causes follicle development retardation and reduces fecundity in zebrafish

Contact Info

Product

Resources

About