青鳉nup58的表达及其在性别分化中的潜在作用

doi:10.16088/j.issn.1001-6600.2023012701

摘要/Abstract

摘要： 核孔复合体是物质穿梭核膜的通道。核孔蛋白能影响核质和胞质间的物质交换,而核质运输具有调控生殖细胞进入雌性或雄性分化途径的能力。性腺体细胞衍生因子(gonadal soma-derived factor, gsdf)能调控青鳉Oryzias latipes雄性生殖细胞,并在有丝分裂最后一轮之前开启或关闭配子发生中起关键作用。通过RT-qPCR实验发现nup58(nucleoporin 58)在野生型青鳉精巢中的表达高于卵巢。对青鳉性腺转录组数据分析发现,nup58在gsdf敲除后精巢中的表达会显著升高。免疫荧光实验结果显示,nup58在配子发生不同阶段的所有生殖细胞中都有表达。我们的研究显示Nup58作为一种核孔蛋白,受gsdf信号的调控,并与雄性生殖细胞的增殖密切相关。由Nup58核孔蛋白协调的核质运输在生殖细胞性别分化中的作用和动态变化值得进一步探讨。

关键词: 青鳉, 性腺, 性别分化, 转录组

Abstract: Nuclear pore complexes (NPCs) form channels to transport materials across the nuclear membrane. Nucleoporin can affect the material exchange between nucleoplasm and cytoplasm, and the nucleoplasm transport has the ability to coordinates germ cells into female or male differentiation pathways. Gonadal soma-derived factor (Gsdf) can regulate the male germ cells of Oryzias latipes and play a key role in switching on/off gametogenesis before the last turn of mitosis. The expression of nup58 (nucleoporin 58) was higher in normal testis than that in normal ovary detected by RT-qPCR. Moreover, compared with normal testicular transcriptome data of medaka(Oryzias latipes), the expression of nup58 in gsdf deficient testis was found to significantly increase by the RNA-seq. The results of immunofluorescence assay showed that nup58 was expressed in all germ cells at different stages of gametogenesis in both wild type and gsdf-knockout medaka gonads. This research shows that Nup58, as a nucleoporin, is regulated by gsdf signal and is closely related to the proliferation of male germ cells. Nup58 participates in dynamic events of nuclear cytoplasmic transport during sexual differentiation, and its synergistic effects need to be explored.

Key words: medaka, gonad, sex differentiation, transcriptome

中图分类号: Q786; S917

李雅园, 王光幸, 李新闻, 郭振华, 关桂君. 青鳉nup58的表达及其在性别分化中的潜在作用[J]. 广西师范大学学报（自然科学版）, 2024, 42(1): 168-179.

LI Yayuan, WANG Guangxing, LI Xinwen, GUO Zhenhua, GUAN Guijun. Expression of nup58 in Medaka and Its Potential Role in Sex Differentiation[J]. Journal of Guangxi Normal University(Natural Science Edition), 2024, 42(1): 168-179.

参考文献

[1] SICINSKI P, DONAHER J L, GENG Y, et al. Cyclin D2 is an FSH-responsive gene involved in gonadal cell proliferation and oncogenesis[J]. Nature, 1996,384(6608):470-474. DOI: 10.1038/384470a0.
[2] DONG P, ZHANG C, PARKER B T, et al. Cyclin D/CDK4/6 activity controls G1 length in mammalian cells[J]. PLoS One, 2018,13(1):e0185637.DOI: 10.1371/journal.pone.0185637.
[3] ALT J R, GLADDEN A B, DIEHL J A. p21^Cip1 promotes cyclin D1 nuclear accumulation via direct inhibition of nuclear export[J]. The Journal of Biological Chemistry, 2002,277(10):8517-8523. DOI: 10.1074/jbc.M108867200.
[4] GASCA S, CANIZARES J, DE SANTA BARBARA P, et al. A nuclear export signal within the high mobility group domain regulates the nucleocytoplasmic translocation of SOX9 during sexual determination[J]. Proceedings of the National Academy of Sciences of the United States of America, 2002,99(17):11199-11204. DOI: 10.1073/pnas.172383099.
[5] PENRAD-MOBAYED M, PERRIN C, L’HÔTE D, et al. A role for SOX9 in post-transcriptional processes: insights from the amphibian oocyte[J]. Scientific Reports, 2018,8(1):7191. DOI: 10.1038/s41598-018-25356-1.
[6] HAMPOELZ B, ANDRES-PONS A, KASTRITIS P, et al. Structure and assembly of the nuclear pore complex[J]. Annual Review of Biophysics, 2019,48:515-536. DOI: 10.1146/annurev-biophys-052118-115308.
[7] AKEY C W, SINGH D, OUCH C, et al. Comprehensive structure and functional adaptations of the yeast nuclear pore complex[J]. Cell, 2022,185(2):361-378. DOI: 10.1016/j.cell.2021.12.015.
[8] LIN D H, HOELZ A. The Structure of the nuclear pore complex (an update)[J]. Annual Review of Biochemistry, 2019,88:725-783. DOI: 10.1146/annurev-biochem-062917-011901.
[9] 王萍, 余自华. 核孔复合体的结构及其功能[J]. 现代生物医学进展, 2019,19(3):566-571. DOI: 10.13241/j.cnki.pmb.2019.03.040.
[10] GOMAR-ALBA M, MENDOZA M. Modulation of cell identity by modification of nuclear pore complexes[J]. Frontiers in Genetics, 2020,10:1301. DOI: 10.3389/fgene.2020.01301.
[11] SHARMA A, SOLMAZ S R, BLOBEL G, et al. Ordered regions of channel nucleoporins Nup62, Nup54, and Nup58 form dynamic complexes in solution[J]. The Journal of Biological Chemistry, 2015,290(30):18370-18378. DOI: 10.1074/jbc.M115.663500.
[12] SONAWANE P J, DEWANGAN P S, MADHESHIYA P K, et al. Molecular and structural analysis of central transport channel in complex with Nup93 of nuclear pore complex[J]. Protein Science, 2020,29(12):2510-2527. DOI: 10.1002/pro.3983.
[13] FORWOOD J K, HARLEY V, JANS D A. The C-terminal nuclear localization signal of the sex-determining region Y (SRY) high mobility group domain mediates nuclear import through importin beta 1[J]. The Journal of Biological Chemistry, 2001,276(49):46575-46582. DOI: 10.1074/jbc.M101668200.
[14] HARLEY V R, LAYFIELD S, MITCHELL C L, et al. Defective importin beta recognition and nuclear import of the sex-determining factor SRY are associated with XY sex-reversing mutations[J]. Proceedings of the National Academy of Sciences of the United States of America, 2003,100(12):7045-7050. DOI: 10.1073/pnas.1137864100.
[15] FOX M S, ARES V X, TUREK P J, et al. Feasibility of global gene expression analysis in testicular biopsies from infertile men[J]. Molecular Reproduction and Development, 2003,66(4):403-421. DOI: 10.1002/mrd.10364.
[16] CHEN Y A, CHOUFANI S, FERREIRA J C, et al. Sequence overlap between autosomal and sex-linked probes on the Illumina HumanMethylation27 microarray[J]. Genomics, 2011,97(4):214-222. DOI: 10.1016/j.ygeno.2010.12.004.
[17] McCARTHY N S, MELTON P E, CADBY G, et al. Meta-analysis of human methylation data for evidence of sex-specific autosomal patterns[J]. BMC Genomics, 2014,15(1):981. DOI: 10.1186/1471-2164-15-981.
[18] 姜玲. 核孔蛋白NPP-3在线虫胚胎染色体空间分布上的功能研究[D]. 哈尔滨:哈尔滨工业大学, 2019.
[19] CHEN F, JIAO X F, ZHANG J Y, et al. Nucleoporin35 is a novel microtubule associated protein functioning in oocyte meiotic spindle architecture[J]. Experimental Cell Research, 2018,371(2): 435-443. DOI: 10.1016/j.yexcr.2018.09.004.
[20] 陈矾. 核膜孔蛋白Nup35和Rae1在小鼠卵母细胞减数分裂成熟过程中的作用和机制研究[D]. 武汉:华中农业大学, 2021.
[21] LI Y P, XU J N, XIONG H, et al. Cancer driver candidate genes AVL9, DENND5A and NUPL1 contribute to MDCK cystogenesis[J]. Oncoscience, 2014,1(12):854-865. DOI: 10.18632/oncoscience.107.
[22] HARTONO, HAZAWA M, LIM K S, et al. Nucleoporin Nup58 localizes to centrosomes and mid-bodies during mitosis[J]. Cell Division, 2019,14: 7. DOI: 10.1186/s13008-019-0050-z.
[23] WANG X, RAMAT A, SIMONELIG M, et al. Emerging roles and functional mechanisms of PIWI-interacting RNAs[J]. Nature Reviews Molecular Cell Biology, 2023,24(2):123-141. DOI: 10.1038/s41580-022-00528-0.
[24] MUNAFÒ M, LAWLESS V R, PASSERA A, et al. Channel nuclear pore complex subunits are required for transposon silencing in Drosophila[J]. eLife, 2021,10:e66321. DOI: 10.7554/eLife.66321.
[25] OKAZAKI R, YAMAZOE K, INOUE Y H. Nuclear export of cyclin B mediated by the Nup62 complex is required for meiotic initiation in Drosophila males[J]. Cells, 2020,9(2): 270. DOI: 10.3390/cells9020270.
[26] ZHANG X, GUAN G J, LI M Y, et al. Autosomal gsdf acts as a male sex initiator in the fish medaka[J]. Scientific Reports, 2016,6:19738. DOI: 10.1038/srep19738.
[27] YANG J Y, YAN R, ROY A X, et al. The I-TASSER Suite: protein structure and function prediction[J]. Nature Methods, 2015,12(1):7-8.DOI: 10.1038/nmeth.3213.
[28] BECK M, HURT E. The nuclear pore complex: understanding its function through structural insight[J]. Nature Reviews Molecular Cell Biology, 2017,18(2):73-89. DOI: 10.1038/nrm.2016.147.
[29] FAGERBERG L, HALLSTRÖM B M, OKSVOLD P, et al. Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics[J]. Molecular & Cellular Proteomics, 2014,13(2):397-406. DOI: 10.1074/mcp.M113.035600.
[30] YU Y, FUSCOE J C, ZHAO C, et al. A rat RNA-Seq transcriptomic BodyMap across 11 organs and 4 developmental stages[J]. Nature Communications, 2014,5:3230. DOI: 10.1038/ncomms4230.
[31] BRAWAND D, WAGNER C E, LI Y I, et al. The genomic substrate for adaptive radiation in African cichlid fish[J]. Nature, 2014,513(7518):375-381. DOI: 10.1038/nature13726.
[32] TZAVLAKI K, MOUSTAKAS A. TGF-β signaling[J]. Biomolecules, 2020,10(3): 487. DOI: 10.3390/biom10030487.
[33] 徐疏梅, 张颖庆, 关桂君. 青鳉(Oryzias latipes)smad3的克隆与表达分析[J]. 基因组学与应用生物学, 2021,40(2):599-606. DOI: 10.13417/j.gab.040.000599.
[34] XU L, KANG Y B, CÖL S, et al. Smad2 nucleocytoplasmic shuttling by nucleoporins CAN/Nup214 and Nup153 feeds TGFbeta signaling complexes in the cytoplasm and nucleus[J]. Molecular Cell, 2002,10(2):271-282. DOI: 10.1016/s1097-2765(02)00586-5.
[35] XU L, ALARCÓN C, CÖL S, et al. Distinct domain utilization by Smad3 and Smad4 for nucleoporin interaction and nuclear import[J]. The Journal of Biological Chemistry, 2003,278(43):42569-42577.DOI: 10.1074/jbc.M307601200.
[36] AVIDOR-REISS T, CARR A, FISHMAN E L. The sperm centrioles[J]. Molecular and Cellular Endocrinology, 2020,518:110987. DOI: 10.1016/j.mce.2020.110987.
[37] KALOUS J, ALESHKINA D. Multiple roles of PLK1 in mitosis and meiosis[J]. Cells, 2023,12(1): 187. DOI: 10.3390/cells12010187.
[38] MARTINO L, MORCHOISNE-BOLHY S, CHEERAMBATHUR D K, et al. Channel nucleoporins recruit PLK-1 to nuclear pore complexes to direct nuclear envelope breakdown in C. elegans[J]. Developmental Cell, 2017,43(2):157-171.DOI: 10.1016/j.devcel.2017.09.019.
[39] LI X, LI X W, LI W H, et al. Sex-specific meiosis responses to Gsdf in medaka (Oryzias latipes)[J]. The FEBS Journal, 2023,290(10): 2760-2779. DOI: 10.1111/febs.16701.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed