Yong Wu, Wen-Lin Chang, Yuan Tian, Ming Ye, Ya-Wei Zhang, Zeng Zhang
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Phylogenetic analysis showed that the D930020B18Rik protein sequence was highly conserved in mammals. Gene set enrichment analysis indicated that D930020B18Rik and its homologous protein might be involved in regulating spermatogenesis of mammals by participating in nucleoplasmic condensation (normalized enrichment score ［NES］ = 1.652, P < 0.01, false discovery rate ［FDR］ = 0.153), meiosis (NES = 1.960, P < 0.01, FDR = 0.001) and formation of microtubule cytoskeleton during mitosis (NES = 1.903, P < 0.01, FDR = 0.009). Dual-luciferase reporter assay revealed that the transcription factors klf5 and foxo1 could identify and bind D930020B18Rik promoters and perform the function of positive or negative transcriptional regulation.</p><p><strong>Conclusion: </strong>The D930020B18Rik gene is expressed in the mouse testis in a time- and location-specific manner, highly associated with spermiogenesis, mainly localized in the nuclei of germ cells, and may be involved in the meiosis of spermatocytes and spermiogenesis.</p>","PeriodicalId":24012,"journal":{"name":"中华男科学杂志","volume":"30 1","pages":"9-17"},"PeriodicalIF":0.0000,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Expression of the D930020B18Rik gene in the mouse testis during spermatogenesis: Characteristics and potential role].\",\"authors\":\"Yong Wu, Wen-Lin Chang, Yuan Tian, Ming Ye, Ya-Wei Zhang, Zeng Zhang\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To investigate the expression pattern of the D930020B18Rik gene in the testis of the mouse in different stages of development and its possible role in spermatogenesis.</p><p><strong>Methods: </strong>Using gene expression profile microarray, we identified highly expressed D930020B18Rik in the mouse testis and analyzed the expression pattern of the gene by qPCR, immunohistochemistry, Western blot and immunofluorescence staining, and verified its function and molecular mechanism using bioinformatics analysis, dual-luciferase reporter assay and cell cycle synchronization.</p><p><strong>Results: </strong>The expression of the D930020B18Rik gene remained low in the testis of the mouse and mainly localized in the cytoplasm of spermatogonia during the first 2 postnatal weeks (PNW), increased from the 3rd PNW to sexual maturity, localized in the cytoplasm of spermatogonia and the nuclei of round and elongated spermatids, but was absent in the nuclei of mature sperm. Phylogenetic analysis showed that the D930020B18Rik protein sequence was highly conserved in mammals. Gene set enrichment analysis indicated that D930020B18Rik and its homologous protein might be involved in regulating spermatogenesis of mammals by participating in nucleoplasmic condensation (normalized enrichment score ［NES］ = 1.652, P < 0.01, false discovery rate ［FDR］ = 0.153), meiosis (NES = 1.960, P < 0.01, FDR = 0.001) and formation of microtubule cytoskeleton during mitosis (NES = 1.903, P < 0.01, FDR = 0.009). 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引用次数: 0
摘要
目的研究D930020B18Rik基因在小鼠睾丸不同发育阶段的表达模式及其在精子发生过程中可能发挥的作用：方法：利用基因表达谱芯片鉴定了小鼠睾丸中高表达的D930020B18Rik基因，并通过qPCR、免疫组化、Western印迹和免疫荧光染色等方法分析了该基因的表达模式，同时利用生物信息学分析、双荧光素酶报告实验和细胞周期同步化等方法验证了该基因的功能和分子机制：结果表明：D930020B18Rik基因在小鼠睾丸中的表达量一直较低，在出生后2周内主要定位于精原细胞的胞浆中，从出生后第3周至性成熟前表达量增加，定位于精原细胞的胞浆和圆形精子及长精子的细胞核中，但在成熟精子的细胞核中没有表达。系统进化分析表明，D930020B18Rik 蛋白序列在哺乳动物中高度保守。基因组富集分析表明，D930020B18Rik及其同源蛋白可能通过参与核质凝聚而参与哺乳动物精子发生的调控（归一化富集得分 ［NES］ = 1.652, P < 0.01, false discovery rate ［FDR］ = 0.153）、减数分裂（NES = 1.960, P < 0.01, FDR = 0.001）和有丝分裂过程中微管细胞骨架的形成（NES = 1.903, P < 0.01, FDR = 0.009）。双荧光素酶报告实验表明，转录因子klf5和foxo1可识别并结合D930020B18Rik启动子，发挥正负转录调控功能：结论：D930020B18Rik基因在小鼠睾丸中的表达具有时间和位置特异性，与精子形成高度相关，主要定位于生精细胞核中，可能参与精母细胞的减数分裂和精子形成。
[Expression of the D930020B18Rik gene in the mouse testis during spermatogenesis: Characteristics and potential role].
Objective: To investigate the expression pattern of the D930020B18Rik gene in the testis of the mouse in different stages of development and its possible role in spermatogenesis.
Methods: Using gene expression profile microarray, we identified highly expressed D930020B18Rik in the mouse testis and analyzed the expression pattern of the gene by qPCR, immunohistochemistry, Western blot and immunofluorescence staining, and verified its function and molecular mechanism using bioinformatics analysis, dual-luciferase reporter assay and cell cycle synchronization.
Results: The expression of the D930020B18Rik gene remained low in the testis of the mouse and mainly localized in the cytoplasm of spermatogonia during the first 2 postnatal weeks (PNW), increased from the 3rd PNW to sexual maturity, localized in the cytoplasm of spermatogonia and the nuclei of round and elongated spermatids, but was absent in the nuclei of mature sperm. Phylogenetic analysis showed that the D930020B18Rik protein sequence was highly conserved in mammals. Gene set enrichment analysis indicated that D930020B18Rik and its homologous protein might be involved in regulating spermatogenesis of mammals by participating in nucleoplasmic condensation (normalized enrichment score ［NES］ = 1.652, P < 0.01, false discovery rate ［FDR］ = 0.153), meiosis (NES = 1.960, P < 0.01, FDR = 0.001) and formation of microtubule cytoskeleton during mitosis (NES = 1.903, P < 0.01, FDR = 0.009). Dual-luciferase reporter assay revealed that the transcription factors klf5 and foxo1 could identify and bind D930020B18Rik promoters and perform the function of positive or negative transcriptional regulation.
Conclusion: The D930020B18Rik gene is expressed in the mouse testis in a time- and location-specific manner, highly associated with spermiogenesis, mainly localized in the nuclei of germ cells, and may be involved in the meiosis of spermatocytes and spermiogenesis.
期刊介绍：
National journal of andrology was founded in June 1995. It is a core journal of andrology and reproductive medicine, published monthly, and is publicly distributed at home and abroad. The main columns include expert talks, monographs (basic research, clinical research, evidence-based medicine, traditional Chinese medicine), reviews, clinical experience exchanges, case reports, etc. Priority is given to various fund-funded projects, especially the 12th Five-Year National Support Plan and the National Natural Science Foundation funded projects. This journal is included in about 20 domestic databases, including the National Science and Technology Paper Statistical Source Journal (China Science and Technology Core Journal), the Source Journal of the China Science Citation Database, the Statistical Source Journal of the China Academic Journal Comprehensive Evaluation Database (CAJCED), the Full-text Collection Journal of the China Journal Full-text Database (CJFD), the Overview of the Chinese Core Journals (2017 Edition), and the Source Journal of the Top Academic Papers of China's Fine Science and Technology Journals (F5000). It has been included in the full text of the American Chemical Abstracts, the American MEDLINE, the American EBSCO, and the database.
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