低压低氧对小鼠精子发生及其小RNA表达的影响
收稿日期: 2021-12-22
录用日期: 2022-07-05
网络出版日期: 2022-09-21
基金资助
国家自然科学基金(81960287);中国科学院青年创新促进会(2021432);青海省重大专项“三江源区代表性动物基因资源保护与应用”
Effects of hypobaric hypoxia on spermatogenesis and the expression of small RNA in mice
Received date: 2021-12-22
Accepted date: 2022-07-05
Online published: 2022-09-21
作为青藏高原最为关键的环境因子，低压低氧对高原非习服动物繁殖和生殖系统功能有不利影响。已有的研究表明，低氧环境会导致雄性生殖细胞凋亡、精子畸形率升高、精子质量下降，进而影响受精和早期胚胎发育，但目前缺乏低氧损伤精子功能的机理研究。小RNA (small RNA) 是在转录后及翻译水平上调控基因表达的重要功能分子，不同类型的small RNA通过诱导基因沉默或调控翻译等方式参与调节精子发生。本研究通过低压氧舱模拟海拔5 000 m处理4周建立缺氧小鼠模型，发现低氧处理导致小鼠曲精细管中生精细胞排列紊乱，精子数量未发生显著变化但畸形率增加17.5倍 (P < 0.001)。通过small RNA测序发现，低氧组小鼠精子中的small RNA碱基偏好性与对照组一致，第一碱基对尿嘧啶 (U) 有很强的偏好性。低氧组小鼠精子中21 nt长度的small RNA比例显著减少4.4% (P < 0.05)。低氧组小鼠精子中piRNA、tsRNA表达无差异，但miRNA表达上调21个，下调58个。对差异miRNAs靶基因与相同低氧处理小鼠睾丸组织差异基因比对，共比对到831个差异表达基因，其中上调miRNAs的靶基因中有429个差异表达基因；下调miRNAs的靶基因中有813个差异表达基因。此外，上调miRNAs的靶基因富集在FoxO信号通路、甲状腺激素信号通路、类固醇生物合成和HIF?1信号等通路，而下调miRNAs的靶基因富集在脂肪酸代谢等通路。本研究获得了缺氧小鼠精子small RNA变化图谱，对人类和其他动物在缺氧环境下精子表观遗传修饰变化的研究具有参考价值。
李双 , 贾功雪 , 陶海萍 , 王玉军 , 李斌业 , 杨其恩 . 低压低氧对小鼠精子发生及其小RNA表达的影响[J]. 兽类学报, 2022 , 42(5) : 579 -589 . DOI: 10.16829/j.slxb.150649
As the most crucial environmental factors, low atmospheric pressure and low oxygen content have adverse effects on the reproductive system of non-adapted animals living in the Qinghai?Tibet Plateau. Compelling evidence has shown that hypoxia exposure caused increased germ cell apoptosis and lower sperm quality, leading to defects in fertilization and preimplantation embryo development. Currently, how hypoxia affects testicular gene expression and sperm function is not well-understood. small RNA regulates gene expression at the post-transcriptional and translational level and participates in spermatogenesis by inducing gene silencing or transcription. In this study, a hypoxia mouse model was established by simulating the environment at an altitude of 5 000 m in a hypobaric oxygen chamber for 4 weeks. The results showed that the germ cells within seminiferous tubules of hypoxic mice were arranged disorderly. Although the total number of sperm did not change significantly, the proportion of abnormal sperm had a 17.5 fold increase in Hypo-4W animals (P < 0.001). small RNA sequencing revealed that the number of small RNAs with a length of 21 nt in hypoxic mouse sperm was reduced by 4.4% (P < 0.05), and the first base had a strong preference for Uracil (U). There were no significant differences in piRNA and tsRNA in sperm of control and hypoxia exposed animals, however, 21 miRNAs were up-regulated and 58 were down-regulated. Furthermore, we analyzed the target genes of these miRNAs and differentially expressed genes in the hypoxia-treated testicular tissues of mice. The results showed that 429 target genes of up-regulated miRNAs and 813 target genes of down-regulated miRNAs were differentially expressed. Finally, enrichment analysis revealed that the target genes of up-regulated miRNAs were enriched in FoxO signaling pathway, thyroid hormone signaling pathway, steroid biosynthesis and HIF-1 signaling pathway, while the targets of down-regulated miRNAs were enriched in fatty acid metabolism. In summary, the miRNA expression dynamics of hypoxic mouse sperm obtained in the present study provides an important reference value for further dissecting changes in sperm epigenome in humans and other animals under hypoxic environment.
Key words： Hypobaric hypoxia; Sperm; small RNA; Target genes
| null | Ayman A H, Anas A O, Yasser A A, Masad A M, Abdullah A S, Mazen A, Salih A G, Saad A S, Abdulraheem A, AbdulRhman A G, Al-Gthami O. 2021. Levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone and prolactin at moderate altitude inhabitant male. Pakistan Journal of Biological Sciences, 24 (2): 188-192. |
| null | Bale T L. 2015. Epigenetic and transgenerational reprogramming of brain development. Nature Reviews Neuroscience, 16 (6): 332-344. |
| null | Chen Q, Yan M H, Cao Z H, Li X, Zhang Y F, Shi J, Feng G H, Peng H Y, Zhang X D, Zhang Y, Qian J J, Duan E K, Zhai Q W, Zhou Q. 2016. Sperm tsRNAs contribute to intergenerational inheritance of an acquired metabolic disorder. Science, 351 (6271): 397-400. |
| null | Chen X X, Li X L, Guo J Y, Zhang P F, Zeng W X. 2017. The roles of microRNAs in regulation of mammalian spermatogenesis. Journal of Animal Science and Biotechnology, 8 (1): 35-42. |
| null | Cheng S K, Yu J Y, Si B H, Xiao Q L, Liang Z J. 2001. Study on the relationship of high hematocrit with changes of the blood viscosity during in the environment of altitude hypoxia. Chinese Journal of Applied Physiology, 17 (3): 231-235. (in Chinese) |
| null | Cui P, Xu D, Xing T, Ren G H, Ma S M. 2020. Relation of ASXL2 gene mutation with clinical characteristics, prognosis and C-KIT gene mutation in AML patients with AML1-ETO fusion gene. Journal of Experimental Hematology, 28 (1): 125-129.(in Chinese) |
| null | Deng C Y, Liu N N, Fan Y Y, Chi H B, Zu L Y.2020. Research progress on the effect and mechanism of altitude hypoxic environment on male semen quality. Chinese Journal of Andrology, 34 (3): 72-75, 79. (in Chinese) |
| null | Dweep H, Gretz N, Sticht C. 2014. miRWalk database for miRNA-target interactions. Methods in Molecular Biology, 1182 (2014):289-305. |
| null | Felli N, Fontana L, Pelosi E, Botta R, Bonci D, Facchiano F, Liuzzi F, Lulli V, Morsilli O, Santoro S, Valtieri M, Calin G A, Liu C G, Sorrentino A, Croce C M, Peschle C. 2005. MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down?modulation. Proceedings of the National Academy of Sciences of the United States of America, 102 (50): 18081-18086. |
| null | Feng E Z, Dai S G, Yang S Y. 2014. Research progress of hypoxic pulmomary artery hypertension. Chinese Journal of Lung Diseases, 7 (3): 84-87. (in Chinese) |
| null | H?pfl G, Ogunshola O, Gassmann M. 2003. Hypoxia and high altitude. The molecular response. Advances in Experimental Medicine and Biology, 543 (1): 89-115. |
| null | Huang F M. 2011. Effects of simulated altitude hypoxia environment at an altitude of 5000 meters on sexual function and behavior of male rats and mice.Master thesis. Beijing: Peking Union Medical College.(in Chinese) |
| null | Huang P, Zhou Z G, Shi F X, Shao G B, Wang R, Wang J T, Wang K X, Ding W. 2016. Effects of the IGF-1/PTEN/Akt/FoxO signaling pathway on male reproduction in rats subjected to water immersion and restraint stress. Molecular Medicine Reports, 14 (6): 5116-5124. |
| null | Huszar J M, Payne C J. 2013. MicroRNA 146 (Mir146) modulates spermatogonial differentiation by retinoic acid in mice. Biology of Reproduction, 88 (1): 1-10, 15. |
| null | Kuramochi?Miyagawa S, Watanabe T, Gotoh K, Totoki Y, Toyoda A, Ikawa M, Asada N, Kojima K, Yamaguchi Y, Ijiri T W, Hata K, Li E, Matsuda Y, Kimura T, Okabe M, Sakaki Y, Sasaki H, Nakano T. 2008. DNA methylation of retrotransposon genes is regulated by Piwi family members MILI and MIWI2 in murine fetal testes. Genes & Development, 22 (7): 908-917. |
| null | Li C H, Yang B, Pan P, Ma Q, Wu Y, Zhang Z, Guo X, Ye J, Gui Y T. 2018. MicroRNA?130a inhibits spermatogenesis by directly targeting androgen receptor in mouse Sertoli cells. Molecular Reproduction & Development, 85 (10): 768-777. |
| null | Li Q F, You Z B, Chen X G, Du J Z. 1986. Effects of chronic hypoxia on the liver of ochotona curzoniae and rats. Acta Theriologica Scnica, 6 (4): 261-266. (in Chinese) |
| null | Li S, Yang Q E. 2022. Hypobaric hypoxia exposure alters transcriptome in mouse testis and impairs spermatogenesis in offspring. Gene, 823: 146390. |
| null | Liao W G, Cai M C, Chen J, Huang J, Liu F Y, Jiang C H, Gao Y Q. 2010. Hypobaric hypoxia causes deleterious effects on spermatogenesis in rats. Reproduction, 139 (6): 1031-1038. |
| null | Liao W G, Gao Y Q, Wu Y, Cai M C, Fan Y M. 2006. Effect of hypoxia on epididymis of rat. Journal of Reproductive Medicine, 15 (4): 252-256. (in Chinese) |
| null | Liu X Y, Liu S Y, Xu C L. 2020. Effects of leptin on HPG axis and reproductive function in male rat in simulated altitude of 5500 m hypoxia environment. Biochemical and Biophysical Research Communications, 529 (1): 104-111. |
| null | Love M I, Huber W, Anders S. 2014. Moderated estimation of fold change and dispersion for RNA?seq data with DESeq2. Genome Biology, 15 (12): 550-570. |
| null | Noblanc A, Damon?Soubeyrand C, Karrich B, Henry?Berger J, Cadet R, Saez F, Guiton R, Janny L, Pons?Rejraji H, Alvarez J G, Drevet J R, Kocer A. 2013. DNA oxidative damage in mammalian spermatozoa: where and why is the male nucleus affected? Free Radical Biology and Medicine, 65 (1): 719-723. |
| null | Nozik?Grayck E, Shimoda L A. 2020. Heart of the Matter: Divergent Roles of HIFs in Hypoxia-induced RV Hypertrophy. American Journal of Respiratory Cell and Molecular Biology, 63 (5):549-550. |
| null | Quarato P, Singh M, Bourdon L, Cecere G. 2022. Inheritance and maintenance of small RNA-mediated epigenetic effects. Bioessays, e2100284. |
| null | Sato Y, Asahina K, Yoshiike M, Nozawa S, Otoi T, Iwamoto T. 2020. A change in the steroid metabolic pathway in human testes showing deteriorated spermatogenesis. Reproductive Biology, 20 (2): 210-219. |
| null | Sharma U, Conine C C, Shea J M, Boskovic A, Derr A G, Bing X Y, Belleannee C, Kucukural A, Serra R W, Sun F, Song L, Carone B R, Ricci E P, Li X Z, Fauquier L, Moore M J, Sullivan R, Mello C C, Garber M, Rando O J. 2016. Biogenesis and function of tRNA fragments during sperm maturation and fertilization in mammals. Science, 351 (6271): 391-396. |
| null | Tao H P, Jia G X, Zhang X N, Wang Y J, Li B Y, Yang Q E. 2021. Paternal hypoxia exposure impairs fertilization process and preimplantation embryo development. Zygote, 23 (1): 1-9. |
| null | Tong H, Zhang M, Wang S, Bai S L.2011. Involvement of sleep intermittent hypoxia in remodeling, apoptosis and expression of Rho kinase in rat myocardial cells. Acta Anatomica Sinica, 42 (4): 470-474. (in Chinese) |
| null | Vlachos I S, Zagganas K, Paraskevopoulou M D, Georgakilas G, Karagkouni D, Vergoulis T, Dalamagas T, Hatzigeorgiou A G. 2015. DIANA?miRPath v3.0: deciphering microRNA function with experimental support. Nucleic Acids Research, 43 (W1): W460-W466. |
| null | Wan L, Wang Y M, Lei J, Lan T, Chang D H, Zhang B. 2012. Expression of CatSperl mRNA and its significance in spermatozoa of healthy males living in high altitude about 6 months. Chinese Journal of Andrology, 26 (12): 27-29, 38. (in Chinese) |
| null | Wang J S, Gong X F, Meng F C, Deng S, Dai H H, Bao B H, Feng J L, Li H S, Wang B. 2020a. Biological network model of effect of chronic intermittent hypoxia on spermatogenesis in rats. Medical Science Monitor, 26 (3): e925579-1-e925579-11. |
| null | Wang X L, Lv C Y, Guo Y, Yuan S Q. 2020b. Mitochondria associated germinal structures in spermatogenesis: piRNA pathway regulation and beyond. Cells, 9 (2): 399-419. |
| null | Wenda J M, Homolka D, Yang Z, Spinelli P, Sachidanandam R, Pandey R R, Pillai R S. 2017. Distinct roles of RNA helicases MVH and TDRD9 in PIWI slicing?triggered mammalian piRNA biogenesis and function. Developmental Cell, 41 (6): 623-637. |
| null | Yamasaki S, Ivanov P, Hu G F, Anderson P. 2009. Angiogenin cleaves tRNA and promotes stress?induced translational repression. Journal of Cell Biology, 185 (1): 35-42. |
| null | Yang Q E, Racicot K E, Kaucher A V, Oatley M J, Oatley J M. 2013. MicroRNAs 221 and 222 regulate the undifferentiated state in mammalian male germ cells. Development, 140 (2): 280-290. |
| null | Yusuke O, Yasushi T, Atsushi T, Toshiaki W, Yasuhiro Y, Katsushi T, Yoshiyuki S, Hiroyuki S, Hirohiko H. 2010. Small RNA class transition from siRNA/piRNA to miRNA during pre?implantation mouse development. Nucleic Acids Research, 38 (15): 5141-5151. |
| null | Zhang L M.2009.Effect of hypoxia stress during pregnancy on reproductive ability of offspring male rats. Master thesis. Hangzhou: Hangzhou Normal University. (in Chinese) |
| null | Zhang N, Yang Q L, Sun Y P. 2016. Role of microRNA in spermatogenesis. Reproduction & Contraception, 35 (1): 37-41. (in Chinese) |
| null | Zhao S, Liu M F. 2010. piRNA and PIWI in animal germ cells. Chinese Bulletin of Life Sciences, 22 (7): 623-627. (in Chinese) |
| null | 万霖, 王养民, 雷静, 蓝天, 常德辉, 张斌. 2012. 移居高原6月后精子CatSperlmRNA的表达及意义. 中国男科学杂志, 26 (12): 27-29, 38. |
| null | 邓琛耀, 刘娜娜, 范媛媛, 迟洪滨, 祖凌云. 2020. 高原低氧环境对男性精液质量影响及其机制的研究进展. 中国男科学杂志, 34 (3): 72-75, 79. |
| null | 冯恩志, 戴胜归, 杨生岳. 2014. 低氧性肺动脉高压研究进展. 中华肺部疾病杂志, 7 (3): 84-87. |
| null | 杜继曾, 张家兴. 2004. 低氧下生长发育抑制与认知功能促进的调节机制. 珠海: 中国生理学会第五届比较生理学学术会议, 16-17. |
| null | 李庆芬, 尤治秉, 陈晓光, 杜继曾. 1986. 慢性高原低氧对高原鼠兔和大鼠肝脏的作用. 兽类学报, 6 (4): 261-266. |
| null | 佟浩, 张曼, 王实, 柏树令. 2011. 睡眠间歇低氧大鼠心室重塑与心肌细胞凋亡和Rho激酶表达的关系. 解剖学报, 42 (4): 470-474. |
| null | 张楠, 杨庆岭, 孙莹璞. 2016. microRNA在精子发生中的作用. 生殖与避孕, 35 (1): 37-41. |
| null | 殷骏, 倪兵, 张梦洁. 2021. ASXL2在高原低氧下调控精子形成基因表达中的应用.专利, CN 113201589A. |
| null | 赵爽, 刘默芳. 2010. piRNA和PIWI蛋白的功能机制研究进展. 生命科学, 22 (7): 623-627. |
| null | 黄付敏. 2011. 模拟海拔5000米高原低氧环境对雄性大小鼠性功能及行为学的影响. 北京: 北京协和医学院硕士学位论文. |
| null | 崔鹏, 徐冬, 邢恬, 任国华, 马尚民. 2020. ASXL2基因突变与伴AML1-ETO融合基因的AML患者的临床特征、预后及C-KIT基因突变的关系.中国实验血液学杂志, 28 (1): 125-129. |
| null | 章丽梅.2009. 孕期低氧应激对子代雄性大鼠生殖能力的影响. 杭州: 杭州师范大学硕士学位论文. |
| null | 程守科, 于军一, 司本辉, 肖庆林, 梁子钧. 2001. 高原低氧环境下红细胞增多和血液粘度间关系的研究. 中国应用生理学杂志, 17 (3): 231-235. |
| null | 廖卫公, 高钰琪, 吴艺, 蔡明春, 范有明. 2006. 低氧对大鼠附睾功能的影响. 生殖医学杂志, 15 (4): 252-256. |
/
| 〈 |
|
〉 |
青公网安备 63010402000199号 青ICP备05000010号-2
