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| Expression Characteristics Analysis of srsf3 Gene in Tissues and Gonad During Development Stages of Chinese Giant Salamander (Andrias davidianus) |
| MENG Fang1,2, FANG Jun-Chao2, TIAN Hai-Feng2, SHAO Peng1*, HU Qiao-Mu2* |
1 College of Aquaculture, Tianjin Agricultural University, Tianjin 300380, China;
2 Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China |
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Abstract There is a significant difference in the growth of male and female giant salamanders (Andrias davidianus), and it has been confirmed that the serine/arginine-rich splicing factor 3 (srsf3) gene is involved in the selective splicing of nuclear receptor subfamily 5 group A member 2 (Nr5a2), a gene related to sex differentiation in giant salamanders. To investigate the mechanism of srsf3 gene in sex differentiation of Chinese giant salamander , qPCR technology was used to analyze the changes in expression levels in different tissues, gonads at different developmental stages, and gonads induced by sex hormones; Fluorescence in situ hybridization (FISH) was used to determine the expression and localization of the srsf3 gene in the testes and ovaries of giant salamanders. Based on the analysis of the transcriptome sequencing results of the giant salamander gonads, the full-length of srsf3 gene was 1 478 bp, with an open reading frame of 504 bp, encoding 167 amino acids. Systematic evolutionary analysis showed that the amino acid sequence of the giant salamander SRSF3 was closely related to the Xenopus tropicalis. The qPCR results showed that the expression level of srsf3 gene was highest in the ovaries of giant salamanders, followed by pituitary tissue, slightly lower in muscle and heart tissues, and low expression in liver, spleen, lung, and stomach tissues. The expression of the srsf3 gene in the gonads of giant salamanders at different stages showed that the ovaries were significantly higher than the testes at ages 2~5 (P<0.05); The expression level of srsf3 gene reached its highest in the ovaries at the age of 2, and then decreased year by year in the ovaries. The expression level tended to stabilize in the ovaries at the age of 4-6. The expression level of the srsf3 gene was highest in sex reversed males, followed by sex reversed females; The expression level of srsf3 gene in the sex reversal ovaries was significantly higher than that in the ovaries (P<0.05), and the expression level in the sex reversal testes was significantly higher than that in the testes (P<0.05). The FISH results showed that the srsf3 gene was expressed in both the ovaries and testes of the giant salamander. The above studies indicated that the srsf3 gene was involved in the gonadal development of giant salamanders and plays an important role in the process of sex reversal in giant salamanders. Functional analysis of the srsf3 gene in this study helps to deepen the understanding of the mechanism of sex differentiation in amphibians.
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Received: 05 July 2024
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Corresponding Authors:
* hqmu0806@163.com;shaopeng@tjau.edu.com
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[1] 蒋瑶, 张荣, 张静怡, 等. 2021.宫颈癌组织YAP1和SRSF3蛋白表达与临床病理参数及预后的关系[J]. 中华实用诊断与治疗杂志, 35: 131-135.(Jiang Y, Zhang R, Zhang J Y, et al.2021. Correlations of the expressions of YAP1 and SRSF3 proteins in cervical cancer tissues withclinicopathological parameters and prognosis[J]. Journal of Chinese Practical Diagnosis and Therapy, 35: 131-135.)
[2] 李成, 江建平, 谢锋, 等. 2023.中国两栖爬行动物多样性监测进展与展望[J]. 生物多样性, 31: 186-198.(Li C, Jiang J P, Xie F, et al.2023. Progress and prospect of Chinese biodiversity monitoring of amphibians and reptiles[J]. Biodiversity Science, 31: 186-198)
[3] 刘淑娟. 2021. miR-486靶向srsf3对奶山羊卵巢颗粒细胞增殖及凋亡调控作用的研究[D]. 硕士学位论文, 西北农林科技大学,导师: 曹斌云, pp.30-42.(Liu S J.2021.Study of mir-486 on the proliferation and apoptosis of goat ovarian granulosa cells by targeting srsf3[D].Thesis for M.S., Northwest A＆F University, Supervisor: Cao B Y, pp. 30-42.)
[4] 王国庆, 特日格乐, 翁雅娟, 等. 2024.剪接体在细胞生命活动领域中的研究进展[J].黑龙江畜牧兽医, (09): 22-29.(Wang G Q, Terigele, Weng Y J, et al. 2024. Research progress of spliceosomes in the field of cell life activities[J]. Heilongjiang Animal Science and Veterinary, (09): 22-29.)
[5] 赵亚楠, 吴思雯, 沙倩倩, 等. 2024. 选择性剪切在动物早期胚胎发育中的作用研究进展[J]. 中国畜牧杂志, 60: 88-92.(Zhao Y N, Wu S W, Sha Q Q,et al.2024. Research progress on the role of alternative splicing in animal early embryonic development[J]. Chinese Journal of Animal Science, 60: 88-92.)
[6] 周璐. 2021. 剪接调控因子SRSF3抑制低氧诱导下自噬的作用和调控机制研究[D]. 硕士学位论文, 武汉大学, 导师: 贾荣, pp. 19-37.(Zhou L.2021. The function and mechanism of splicing factor SRSF3 in hypoxia-induced autophagy[D].Thesis for M.S., Wuhan University,Supervisor: Jia R, pp. 19-37.)
[7] ÄNKö M L.2014. Regulation of gene expression programmes by serine-arginine rich splicing factors[J]. Seminars in Cell & Developmental Biology, 32: 11-21.
[8] Bonnal S C, López I, Valcárcel J.2020. Roles and mechanisms of alternative splicing in cancer-implications for care[J]. Nature Reviews Clinical Oncology, 17: 457-474.
[9] Che Y, Bai M, Lu K, et al.2023. Splicing factor SRSF3 promotes the progression of cervical cancer through regulating DDX5[J]. Molecular Carcinogenesis, 62: 210-223.
[10] Conrad T, Akhtar A.2012. Dosage compensation in Drosophila melanogaster: Epigenetic fine-tuning of chromosome-wide transcription[J]. Nature Reviews Genetics, 13: 123-134.
[11] Du C, McGuffin M E, Dauwalder B, et al.1998. Protein phosphorylation plays an essential role in the regulation of alternative splicing and sex determination in Drosophila[J]. Molecular Cell, 2(6): 741-750.
[12] Ellegren H, Hultin L, Brunström B, et al.2007. Faced with inequality: Chicken do not have a general dosage compensation of sex-linked genes[J]. BMC Biology, 5: 40.
[13] Fu X D.1993. Specific commitment of different pre-mRNAs to splicing by single SR proteins[J]. Nature, 365: 82-85.
[14] Fu X D, Maniatis T.1992. The 35-kDa mammalian splicing factor SC35 mediates specific interactions between U1 and U2 small nuclear ribonucleoprotein particles at the 3' splice site[J]. Proceedings of the National Academy of Sciences of the USA, 89: 1725-1729.
[15] Gross T, Richert K, Mierke C, et al.1998. Identification and characterization of srp1, a gene of fission yeast encoding a RNA binding domain and a RS domain typical of SR splicing factors[J]. Nucleic Acids Research, 26: 505-511.
[16] Heinrichs V, Baker B S.1995. The Drosophila SR protein RBP1 contributes to the regulation of doublesex alternative splicing by recognizing RBP1 RNA target sequences[J]. The EMBO Journal, 14: 3987-4000.
[17] Hu Q M, Chang C, Wang Q, et al.2019a. Genome-wide RAD sequencing to identify a sex-specific marker in Chinese giant salamander Andrias davidianus[J]. BMC Genomics, 20(1): 415.
[18] Hu Q M, Liu Y, Liao X L, et al.2021. A high-density genetic map construction and sex-related loci identification in Chinese Giant salamander[J]. BMC Genomics, 22: 230.
[19] Hu Q M, Meng Y, Wang D, et al.2019b. Characterization and function of the T-box 1 gene in Chinese giant salamander Andrias davidianus[J]. Genomics, 111(6): 1351-1359.
[20] Hu Q M, Tian H F, Li W, et al.2019c. Identification of critical sex-biased genes in Andrias davidianus by de novo transcriptome[J]. Molecular Genetics & Genomics, 294: 287-299.
[21] Hu Q M, Tian H F, Xiao H B.2019d. Effects of temperature and sex steroids on sex ratio, growth, and growth-related gene expression in the Chinese giant salamander Andrias davidianus[J]. Aquatic Biology, 28: 79-90.
[22] Kano S, Nishida K, Kurebe H, et al.2014. Oxidative stress-inducible truncated serine/arginine-rich splicing factor 3 regulates interleukin-8 production in human colon cancer cells[J]. American Journal of Physiology-Cell Physiology, 306: C250-C262.
[23] Krainer A R, Conway G C, Kozak D1990. Purification and characterization of pre-mRNA splicing factor SF2 from HeLa cells[J]. Genes & Development, 4: 1158-1171.
[24] Krzywinska E, Ferretti L, Li J, et al.2021. Femaleless controls sex determination and dosage compensation pathways in females of Anopheles mosquitoes[J]. Current Biology, 31(5): 1084-1091.
[25] Lucchesi J C, Kuroda M I.2015. Dosage compensation in Drosophila[J]. Cold Spring Harbor Perspectives in Biology, 7(5): a019398.
[26] Manolakou P, Lavranos G, Angelopoulou R.2006. Molecular patterns of sex determination in the animal kingdom: A comparative study of the biology of reproduction[J]. Reproductive Biology and Endocrinology, 4: 59.
[27] Ohno D S.1967. Sex Chromosomes and Sex-linked Genes[M].Springer-Verlag Berlin Heidelberg, German, pp.101-112.
[28] Sharma A, Heinze S D, Wu Y, et al.2017. Male sex in houseflies is determined by Mdmd, a paralog of the generic splice factor gene CWC22[J]. Science, 356: 642-645.
[29] Sternburg E L, Karginov F V.2020. Global approaches in studying RNA-binding protein interaction networks[J]. Trends in Biochemical Sciences, 45: 593-603.
[30] Yuan S, Feng S, Li J, et al.2021.hnRNPH1 recruits PTBP2 and SRSF3 to modulate alternative splicing in germ cells[J]. Nature Communications,13: 3588-3590.
[31] Zhang J K, Xia X P, Zhu Y, et al.2022. Potential antagonisticrelationship of fgf9 and rspo1 genes in WNT4 pathway to regulate the sex differentiation in Chinese giant salamander (Andrias davidianus)[J].Frontiers in Molecular Biosciences, 9: 974348. |
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