动物性别控制技术研究进展

doi:10.11843/j.issn.0366-6964.2024.04.003

摘要/Abstract

摘要： 性别控制是通过人为干预使雌性动物生产出特定性别后代的一项技术。鉴于X精子和Y精子的分离是实现动物性别控制的技术基础，本文基于X精子和Y精子在物理和化学上的特性差异总结和比较了受精前分离精子的新技术，包括流式细胞分离法、免疫学分离法、pH分离法、微流体和纳米技术。免疫学分离法中R848(雷西莫特)激活TLR7/8信号通路是精子分离的潜在方法，旨在为动物性别控制技术提供理论依据。

关键词: 性别控制, X精子, Y精子, 免疫学分离法, TLR7/8

Abstract: Sex control is a technique in which a female animal produces offspring with a specific sex through human intervention. Since the separation of X sperm and Y sperm is the technical basis for the realization of sperm sexing in animals, this paper summarizes and compares the new techniques for the separation of sperm before fertilization, including flow cytometry, immunological approach of sperm sexing, pH separation, microfluidics and nanotechnology. The activation of TLR7/8 signaling pathway by R848 (Resiquimod) in immunological isolation is a potential method for sperm isolation, aiming to provide a theoretical basis for sperm sexing techniques.

Key words: sperm sexing, X sperm, Y sperm, immunological approach of sperm sexing, TLR7/8

中图分类号:

S814

陈虹宇, 魏雅婷, 李若玺, 高留涛, 刘深贺. 动物性别控制技术研究进展[J]. 畜牧兽医学报, 2024, 55(4): 1370-1380.

CHEN Hongyu, WEI Yating, LI Ruoxi, GAO Liutao, LIU Shenhe. Advances in Animals Sperm Sexing Techniques[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1370-1380.

参考文献

[1] MANZOOR A, PATOO R A, AKRAM T, et al. Sperm sexing and its utility in commercial cattle production:a review[J]. Adv Anim Vet Sci, 2017, 5(7):293-298.
[2] GARNER D L. Flow cytometric sexing of mammalian sperm[J]. Theriogenology, 2006, 65(5):943-957.
[3] JOHNSON L A. Sexing mammalian sperm for production of offspring:the state-of-the-art[J]. Anim Reprod Sci, 2000, 60-61:93-107.
[4] ESPINOSA-CERVANTES R, CÓRDOVA-IZQUIERDO A. Sexing sperm of domestic animals[J]. Trop Anim Health Prod, 2013, 45(1):1-8.
[5] JOHNSON L A, WELCH G R. Sex preselection:high-speed flow cytometric sorting of X and Y sperm for maximum efficiency[J]. Theriogenology, 1999, 52(8):1323-1341.
[6] VISHWANATH R, MORENO J F. Semen sexing-current state of the art with emphasis on bovine species[J]. Animal, 2018, 12(S1):s85-s96.
[7] GAUR P, SAINI G, SAHARAN P, et al. Sex sorted semen-methods, constraints and future perspective[J]. Vet Res Int, 2020, 8(4):368-375.
[8] NAIDU S J, ARANGASAMY A, SELVARAJU S, et al. Spermatozoa sorting techniques for the sex pre-selection:a review[J]. Indian J Anim Res, 2021, B(4530):1-10.
[9] SHARPE J C, EVANS K M. Advances in flow cytometry for sperm sexing[J]. Theriogenology, 2009, 71(1):4-10.
[10] NECULAI-VALEANU A S, ARITON A M. Game-changing approaches in sperm sex-sorting:microfluidics and nanotechnology[J]. Animals(Basel), 2021, 11(4):1182.
[11] HE Q F, WU S H, HUANG M, et al. Effects of diluent pH on enrichment and performance of dairy goat X/Y sperm[J]. Front Cell Dev Biol, 2021, 9:747722.
[12] GARNER D L, EVANS K M, SEIDEL G E. Sex-sorting sperm using flow cytometry/cell sorting[M]//CARRELL D T, ASTON K I. Spermatogenesis:Methods and Protocols. Totowa:Humana Press, 2013:279-295.
[13] GONZÁLEZ-MARÍN C, GÓNGORA C E, MORENO J F, et al. Small ruminant SexedULTRA^TM sperm sex-sorting:status report and recent developments[J]. Theriogenology, 2021, 162:67-73.
[14] GONZÁLEZ-MARÍN C, GÓNGORA C E, GILLIGAN T B, et al. In vitro sperm quality and DNA integrity of SexedULTRA^TM sex-sorted sperm compared to non-sorted bovine sperm[J]. Theriogenology, 2018, 114:40-45.
[15] REN F, XI H M, REN Y J, et al. TLR7/8 signalling affects X-sperm motility via the GSK3α/β-hexokinase pathway for the efficient production of sexed dairy goat embryos[J]. J Anim Sci Biotechnol, 2021, 12(1):89.
[16] XIE Y S, XU Z Q, WU Z F, et al. Sex manipulation technologies progress in livestock:a review[J]. Front Vet Sci, 2020, 7:481.
[17] PRASAD S, RANGASAMY S, SATHESHKUMAR S. Sex preselection in domestic animals-current status and future prospects[J]. Vet World, 2010, 3(7):346-348.
[18] XIONG X R, ZHANG Y, XIONG Y, et al. The establishment of X, Y sperm sorting system in yak by flow cytometer[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2):399-407. (in Chinese)
熊显荣, 张雁, 熊燕, 等. 利用流式细胞仪建立牦牛X、Y精子分选体系的研究[J]. 畜牧兽医学报, 2021, 52(2):399-407.
[19] POZDYSHEV D V, KOMBAROVA N A, MURONETZ V I. Biochemical features of X or Y chromosome-bearing spermatozoa for sperm sexing[J]. Biochemistry (Mosc), 2023, 88(5):655-666.
[20] RAHMAN M S, PANG M G. New biological insights on X and Y chromosome-bearing spermatozoa[J]. Front Cell Dev Biol, 2020, 7:388.
[21] YADAV S K, GANGWAR D K, SINGH J, et al. An immunological approach of sperm sexing and different methods for identification of X- and Y-chromosome bearing sperm[J]. Vet World, 2017, 10(5):498-504.
[22] YATA V K. Recent advances and challenges in the development of novel sperm sexing methods[M]//YATA V K. Sperm Sexing and Its Role in Livestock Production. Singapore:Springer, 2022:65-81.
[23] WU C H, CHEN W D, YANG W Z, et al. Expression of Toll-like receptors 7/8 in Boar reproductive system and evaluation of effect of their ligands on boar X/Y sperm sorting[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4):1490-1499. (in Chinese)
吴昌华, 陈伟东, 杨文政, 等. Toll样受体7/8在公猪生殖系统中的表达及其配体对猪X/Y精子分选效果评价[J]. 畜牧兽医学报, 2023, 54(4):1490-1499.
[24] MA S J, XU Y J, HE K, et al. Molecular evolution and expression patterns of a multigene family of Toll-like receptors in ruminants[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9):3722-3734. (in Chinese)
马淑娟, 徐祎洁, 何珂, 等. 反刍动物Toll样受体多基因家族的分子进化及表达模式分析[J]. 畜牧兽医学报, 2023, 54(9):3722-3734.
[25] YATA V K, SINGH S K, KUMAR S, et al. Use of sexed semen for genetic improvement of indigenous dairy cattle and buffaloes productivity[J]. Indian J Anim Sci, 2022, 92(7):797-805.
[26] KRAWCZYK C M, HOLOWKA T, SUN J, et al. Toll-like receptor-induced changes in glycolytic metabolism regulate dendritic cell activation[J]. Blood, 2010, 115(23):4742-4749.
[27] UMEHARA T, TSUJITA N, SHIMADA M. Activation of Toll-like receptor 7/8 encoded by the X chromosome alters sperm motility and provides a novel simple technology for sexing sperm[J]. PLoS Biol, 2019, 17(8):e3000398.
[28] EVERTS B, AMIEL E, HUANG S C C, et al. TLR-driven early glycolytic reprogramming via the kinases TBK1-IKK? supports the anabolic demands of dendritic cell activation[J]. Nat Immunol, 2014, 15(4):323-332.
[29] WEN F, LIU W D, LI Y, et al. TLR7/8 agonist (R848) inhibit bovine X sperm motility via PI3K/GSK3α/β and PI3K/NFκB pathways[J]. Int J Biol Macromol, 2023, 232:123485.
[30] XU Y L, FAN Y, FAN W M, et al. RNASET2 impairs the sperm motility via PKA/PI3K/calcium signal pathways[J]. Reproduction, 2018, 155(4):383-392.
[31] ZHU X X, SHI D Y, LI X Q, et al. TLR signalling affects sperm mitochondrial function and motility via phosphatidylinositol 3-kinase and glycogen synthase kinase-3α[J]. Cell Signal, 2016, 28(3):148-156.
[32] UMEHARA T, TSUJITA N, ZHU Z D, et al. A simple sperm-sexing method that activates TLR7/8 on X sperm for the efficient production of sexed mouse or cattle embryos[J]. Nat Protoc, 2020, 15(8):2645-2667.
[33] LIBERT C, DEJAGER L, PINHEIRO I. The X chromosome in immune functions:when a chromosome makes the difference[J]. Nat Rev Immunol, 2010, 10(8):594-604.
[34] REN F. The research on key molecular mining of spermatogenesis and sex-sorted semen in dairy goats[D]. Yangling:Northwest A&F University, 2021. (in Chinese)
任发. 奶山羊精子发生关键分子挖掘与性控精液研究[D]. 杨凌:西北农林科技大学, 2021.
[35] KWON W S, RAHMAN M S, LEE J S, et al. A comprehensive proteomic approach to identifying capacitation related proteins in boar spermatozoa[J]. BMC Genomics, 2014, 15(1):897.
[36] JOSHI H, MATHUR M, MOHANTY A, et al. Semen sexing in bovine:current status and the need to develop alternative techniques[J]. Indian J Anim Sci, 2021, 1(1):17-31.
[37] SOLEYMANI B, PARVANEH S, MOSTAFAIE A. Goat polyclonal antibody against the sex determining region Y to Separate X- and Y-chromosome bearing spermatozoa[J]. Rep Biochem Mol Biol, 2019, 8(3):326-334.
[38] SOLEYMANI B, MANSOURI K, RASTEGARI-POUYANI M, et al. Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm[J]. Reprod Domest Anim, 2021, 56(2):270-277.
[39] CHOWDHURY M M R, XU L G, KONG R, et al. In vitro production of sex preselected cattle embryos using a monoclonal antibody raised against bull sperm epitopes[J]. Anim Reprod Sci, 2019, 205:156-164.
[40] CHEN X L, ZHU H B, WU C J, et al. Identification of differentially expressed proteins between bull X and Y spermatozoa[J]. J Proteomics, 2012, 77:59-67.
[41] DE CANIO M, SOGGIU A, PIRAS C, et al. Differential protein profile in sexed bovine semen:shotgun proteomics investigation[J]. Mol Biosyst, 2014, 10(6):1264-1271.
[42] SCOTT C, DE SOUZA F F, ARISTIZABAL V H V, et al. Proteomic profile of sex-sorted bull sperm evaluated by SWATH-MS analysis[J]. Anim Reprod Sci, 2018, 198:121-128.
[43] ZHANG J, HE D B, LIU C L, et al. Differential expression proteomics between X and Y spermatozoa of bull[J]. Journal of Inner Mongolia University:Natural Science Edition, 2023, 54(1):80-90. (in Chinese)
张洁, 何定波, 刘春丽, 等. 牛X与Y精子差异蛋白质组学研究[J]. 内蒙古大学学报:自然科学版, 2023, 54(1):80-90.
[44] BYRNE K, LEAHY T, MCCULLOCH R, et al. Comprehensive mapping of the bull sperm surface proteome[J]. Proteomics, 2012, 12(23-24):3559-3579.
[45] LI S F. The membrane protein differences of X/Y spermatozoa in dairy cattle and dairy goats[D]. Hohhot:Inner Mongolia University, 2021. (in Chinese)
李淑芳. 奶牛及奶山羊X、Y精子膜蛋白差异分析[D]. 内蒙古:内蒙古大学, 2021.
[46] SHEN D, ZHOU C H, CAO M Y, et al. Differential membrane protein profile in bovine X-and Y-Sperm[J]. J Proteome Res, 2021, 20(6):3031-3042.
[47] LAXMIVANDANA R, PATOLE C, SHARMA T R, et al. Differential proteins associated with plasma membrane in X-and/or Y-chromosome bearing spermatozoa in indicus cattle[J]. Reprod Domest Anim, 2021, 56(6):928-935.
[48] OYEYIPO I P, VAN DER LINDE M, DU PLESSIS S S. Environmental exposure of sperm sex-chromosomes:a gender selection technique[J]. Toxicol Res, 2017, 33(4):315-323.
[49] WURLINA W, HARIADI M U, SAFITRI E, et al. The effect of crude guava leaf tannins on motility, viability, and intact plasma membrane of stored spermatozoa of Etawa crossbred goats[J]. Vet World, 2020, 13(3):530-537.
[50] MUEHLEIS P M, LONG S Y. The effects of altering the pH of seminal fluid on the sex ratio of rabbit offspring[J]. Fertil Steril, 1976, 27(12):1438-1445.
[51] YOU Y A, KWON W S, SAIDUR RAHMAN M, et al. Sex chromosome-dependent differential viability of human spermatozoa during prolonged incubation[J]. Hum Reprod, 2017, 32(6):1183-1191.
[52] CONTRI A, GLORIA A, ROBBE D, et al. Kinematic study on the effect of pH on bull sperm function[J]. Anim Reprod Sci, 2013, 136(4):252-259.
[53] RIZVI A A, QURAISHI M I, SARKAR V, et al. The effect of pH and viscosity on bovine spermatozoa motility under controlled conditions[J]. Int Urol Nephrol, 2009, 41(3):523-530.
[54] HE Q F, WU S H, GAO F, et al. Diluent pH affects sperm motility via GSK3α/β-hexokinase pathway for the efficient enrichment of X-sperm to increase the female kids rate of dairy goats[J]. Theriogenology, 2023, 201:1-11.
[55] WONGTAWAN T, DARARATANA N, OONKHANOND B, et al. Zeta potential of bovine X and Y sperm and its application for sperm sorting[J]. Thai J Vet Med, 2018, 48(Suppl):O25-O26.
[56] PHIPHATTANAPHIPHOP C, LEKSAKUL K, WANTA T, et al. Antibody-conjugated magnetic beads for sperm sexing using a multi-wall carbon nanotube microfluidic device[J]. Micromachines (Basel), 2022, 13(3):426.
[57] DOMÍNGUEZ E, MORENO-IRUSTA A, CASTEX H R, et al. Sperm sexing mediated by magnetic nanoparticles in donkeys, a preliminary in vitro study[J]. J Equine Vet Sci, 2018, 65:123-127.
[58] BURINARU T A, AVRAM M, AVRAM A, et al. Detection of circulating tumor cells using microfluidics[J]. ACS Comb Sci, 2018, 20(3):107-126.
[59] LEE D, HWANG B, KIM B. The potential of a dielectrophoresis activated cell sorter (DACS) as a next generation cell sorter[J]. Micro Nano Syst Lett, 2016, 4(1):2.
[60] ZHANG Y L, CHEN X Y. Dielectrophoretic microfluidic device for separation of red blood cells and platelets:a model-based study[J]. J Braz Soc Mech Sci Eng, 2020, 42(2):89.
[61] YANG P H, SUN X S, CHIU J F, et al. Transferrin-mediated gold nanoparticle cellular uptake[J]. Bioconjug Chem, 2005, 16(3):494-496.
[62] GAMRAD L, MANCINI R, WERNER D, et al. Triplex-hybridizing bioconjugated gold nanoparticles for specific Y-chromosome sequence targeting of bull spermatozoa[J]. Analyst, 2017, 142(11):2020-2028.
[63] WONGTAWAN T, DARARATANA N, THONGKITTIDILOK C, et al. Enrichment of bovine X-sperm using microfluidic dielectrophoretic chip:a proof-of-concept study[J]. Heliyon, 2020, 6(11):e05483.
[64] DARARATANA N, TUANTRANONT A, WONGTAWAN T, et al. The dielectrophoresis microfluidic chip for cell separation:case study of separation of floating cell and moving cells[C]//Proceedings of the 8th Biomedical Engineering International Conference. Pattaya:IEEE, 2015:1-5.
[65] QUELHAS J, SANTIAGO J, MATOS B, et al. Bovine semen sexing:sperm membrane proteomics as candidates for immunological selection of X-and Y-chromosome-bearing sperm[J]. Vet Med Sci, 2021, 7(5):1633-1641.
[66] SRINGARM K, THONGKHAM M, MEKCHAY S, et al. High-efficiency bovine sperm sexing used magnetic-activated cell sorting by coupling scFv antibodies specific to Y-chromosome-bearing sperm on magnetic microbeads[J]. Biology (Basel), 2022, 11(5):715.
[67] LI X M, YI Z R, ZHANG P Y, et al. Comparative analysis of the application effect of sexually controlled frozen semen and ordinary frozen semen in holstein dairy cows[J]. China Feed, 2023(8):38-41. (in Chinese)
李雪梅, 易宗容, 张平英, 等. 荷斯坦奶牛性控冻精与常规冻精应用效果比较分析[J]. 中国饲料, 2023(8):38-41.
[68] HUANG M, CAO X Y, HE Q F, et al. Alkaline semen diluent combined with R848 for separation and enrichment of dairy goat X-sperm[J]. J Dairy Sci, 2022, 105(12):10020-10032.