INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2026
·Vol. 27
·期 19
·8588
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IF 5.6
Junqi Qin,Yueying Deng,Yuntao Lu,Yang Zhang,Luping Liu,Qing Luo,Haiyang Liu,Shuzhan Fei,Yuandong Sun,Mi Ou
Anti-Müllerian hormone (Amh) is a key regulator of sex determination and differentiation in fish. In this study, we identified and characterized a single-copyAmhortholog, designatedCmAmh, in blotched snakehead (Channa maculata), an economically important species in China. The cDNA spans 2340 bp and encodes a 545-amino-acid protein with conserved AMH-N and TGF-β domains.CmAmhexhibited pronounced male-biased gonadal expression, with transcript levels in testes markedly exceeding those in ovaries across all developmental stages (45~365 days post-hatching (dph)). Cellular localization revealedCmAmhtranscripts in the Sertoli cells and spermatogonia of the testis and, more weakly, in the oogonia and primary oocytes of the ovary, suggesting a primary role in spermatogenesis and a potential modulatory function in early oogenesis. A strong inverse correlation between promoter CpG methylation and transcriptional activity, hypermethylation in ovaries (82.5~89.7%) versus hypomethylation (15.8~19.1%) in testes, indicated epigenetic regulation, which functionally integrates with hormonal signaling: 17β-estradiol (E2) suppressed, whereas methyltestosterone (MT) upregulatedCmAmhexpression. Long-term E2exposure induced male-to-female secondary sex reversal (SSR) in 26.7% of XY individuals, whereas MT failed to reverse XX fish, indicating thatCmAmhmodulation alone is insufficient to override ovarian fate. Furthermore, CRISPR/Cas9-mediated mutagenesis achieved 40~45% editing efficiency, predominantly generating frameshift alleles, providing a platform for future functional studies. Collectively, these findings positionCmAmhas a hormone-responsive nodal point within a broader sex-differentiation network. This work provides a theoretical foundation for sex-control strategies inC. maculataaquaculture, while future studies should focus on homozygous mutant generation to definitively establish its loss-of-function phenotype and regulatory networks.
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DOI
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 3
·1740
·
IF 4.9
Yonglin Ruan,Xuehui Li,Gang Zhai,Qiyong Lou,Xia Jin,Jiangyan He,Zhan Yin
The mechanism of fish gonadal sex differentiation is complex and regulated by multiple factors. It has been widely known that proper steroidogenesis in Leydig cells and sex-related genes in Sertoli cells play important roles in gonadal sex differentiation. In teleosts, the precise interaction of these signals during the sexual fate determination remains elusive, especially their effect on the bi-potential gonad during the critical stage of sexual fate determination. Recently, all-testis phenotypes have been observed in thecyp17a1-deficient zebrafish and common carp, as well as incyp19a1a-deficient zebrafish. By matingcyp17a1-deficient fish with transgenic zebrafishTg(piwil1:EGFP-nanos3UTR), germ cells in the gonads were labelled with enhanced green fluorescent protein (EGFP). We classified thecyp17a1-deficient zebrafish and their control siblings into primordial germ cell (PGC)-rich and -less groups according to the fluorescence area of the EGFP labelling. Intriguingly, the EGFP-labelled bi-potential gonads incyp17a1+/+ fish from the PGC-rich group were significantly larger than those of thecyp17a1−/− fish at 23 days post-fertilization (dpf). Based on the transcriptome analysis, we observed that thecyp17a1-deficient fish of the PGC-rich group displayed a significantly upregulated expression ofamhandgsdfcompared to that of control fish. Likewise, the upregulated expressions ofamhandgsdfwere observed incyp19a1a-deficient fish as examined at 23 dpf. This upregulation ofamhandgsdfcould be repressed by treatment with an exogenous supplement of estradiol. Moreover, tamoxifen, an effective antagonist of both estrogen receptor α and β (ERα and Erβ), upregulates the expression ofamhandgsdfin wild-type (WT) fish. Using thecyp17a1- andcyp19a1a-deficient zebrafish, we provide evidence to show that the upregulated expression ofamhandgsdfdue to the compromised estrogen signaling probably determines their sexual fate towards testis differentiation. Collectively, our data suggest that estrogen signaling inhibits the expression ofamhandgsdfduring the critical time of sexual fate determination, which may broaden the scope of sex steroid hormones in regulating gonadal sex differentiation in fish.
研究领域:
发育生物学
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内分泌学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
ENVIRONMENT INTERNATIONAL
·
2020
·
IF 7.577
Miaoling Lin,Rui Hua,Jing Ma,Yao Zhou,Pei Li,Xiya Xu,Zhiqiang Yu,Song Quan
Background Bisphenol A (BPA) is a widespread endocrine-disrupting chemical with estrogen like effects, which could interfere with the human reproductive system by disrupting the normal function of granulosa cells (GCs) leading to abnormal ovarian function. However, the mechanism of its toxicity on human GCs has not been clearly described thus far. Methods 106 normogonadotropic infertile women undergoing their first in-vitro fertilization-embryo transfer (IVF-ET) cycle were recruited. Urinary BPA level and the early outcomes of IVF-ET were analysed. Patients were divided to low and high BPA exposure groups using the median urinary BPA concentration as the cut-off value. In-vivo and in-vitro studies were conducted using mice and human granulosa cell line (KGN cells). Female Kunming mice approximately 6–8 weeks of age were poisoned with BPA at different dosages (1, 10 or 100 μg/kg) by oral gavage once daily for 2 weeks, while KGN cells were exposed to BPA at the concentration of 1, 10 or 100 nM for 24 h, 48 h or 72 h. BPA-induced ovarian morphologic changes were analysed by histopathology investigation. Cell viability and apoptosis were evaluated using CCK-8, TUNEL and flowcytometric, respectively. Hormone levels were determined using ELISA and the molecular mechanism studies were conducted using immunofluorescence, RT-PCR and western blots. Results The oocyte retrieval rate, maturation rate and embryo implantation rate significantly decreased with the higher level of urinary BPA concentration. Peak E2 level was lower in high BPA group, but no statistical significance could be observed. In BPA treated mice, cystic dilation of the follicles with a decreased number of GCs could be observed histopathologically. Decreased E2, P4 and AMH level and GCs autophagy could be detected both in-vivo and in-vitro with the activation of AMPK/mTOR/ULK1 signalling pathway. As being confirmed in KGN cells, phosphorylated AMPK and ULK1 increased while phosphorylated mTOR decreased, and by inhibition autophagy using knockdown of AMPK or 3-MA, adverse effects of BPA exposure in-vitro could be reversed. Conclusion BPA exposure might abnormally influence human ovarian functions leading to abnormal folliculogenesis by activation of autophagy in GCs through AMPK/mTOR/ULK1 pathway.
研究领域:
内分泌学
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生殖生物学
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毒理学
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细胞生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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