绵羊精子细胞核质转运蛋白KPNA4的研究

doi:10.11843/j.issn.0366-6964.2022.07.016

Abstract

Abstract: The study aimed to analyze the protein expression of sperm in epididymis at 3 different regions (caput, corpus and cauda) and obtain differentially expressed proteins using proteomics in sheep. The functional enrichment analysis on the data was performed to explore proteins regulating spermatogenesis and maturation. In this study, 3 healthy male Hu sheep around 12 months of age were selected as experimental animals, the epididymis were isolated and sperms were collected by regions. There were 3 groups of samples (caput group, corpus group and cauda group), 3 biological replicates in each group, totaling 9 samples of sheep sperm. Based on TMT-calibrated quantitative protein technology and R software, GO and KEGG enrichment analysis were performed on differentially expressed proteins, and the reliability of the results was verified by Western blot, immunofluorescence and flow cytometry. A total of 616 differential proteins were identified from 22 841 unique peptides. Among them, cauda vs. caput group had 309 differentially expressed proteins(up:213, down:96), cauda vs corpus group had 167 differentially expressed proteins(up:107, down:60), corpus vs caput group had 140 differentially expressed proteins(up:88, down:52). According to the fold change and protein function, the key protein KPNA4, which may be related to sperm maturation and nuclear material transport, was screened out. This study reveals the characteristics and differences of sperm from different regions of the sheep epididymis. These data provide a rich resources for studying the reproductive mechanism and sperm maturation in male sheep.

Key words: sheep, proteomics, sperm maturation

CLC Number:

S826.3

YAN Shuo, ZHAO Shanshan, ZHU Zhendong, PAN Qingjie, DONG Huansheng. Study on Nuclear-plasma Transporter KPNA4 of Sheep Sperm Cell[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2194-2201.

References 35

[1]	BRIZ M D,BONET S,PINART B,et al.Comparative study of boar sperm coming from the caput,corpus,and cauda regions of the epididymis[J].J Androl,1995,16(2):175-188.
[2]	CHOCU S,CALVEL P,ROLLAND A D,et al.Spermatogenesis in mammals:proteomic insights[J].Syst Biol Reprod Med,2012, 58(4):179-190.
[3]	JAMES E R,CARRELL D T,ASTON K I,et al.The role of the epididymis and the contribution of epididymosomes to mammalian reproduction[J].Int J Mol Sci,2020,21(15):5377.
[4]	OZKOCER S E,KONAC E.The current perspective on genetic and epigenetic factors in sperm maturation in the epididymis[J]. Andrologia,2021,53(3):e13989.
[5]	LIU J M,YAO T,WENG X X,et al.Antioxidant properties and transcriptome of cauda epididymis with different levels of fertility in Hu lambs[J].Theriogenology,2022,182:85-95.
[6]	NIXON B,DE IULIIS G N,HART H M,et al.Proteomic profiling of mouse epididymosomes reveals their contributions to post-testicular sperm maturation[J].Mol Cell Proteomics,2019,18(S1):S91-S108.
[7]	TINGARI M.The fine structure of the epithelial lining of the epididymis of the camel (Camelus dromedarius) with special reference to regional differences[J].J Anat,1989,165:201-204.
[8]	CHU C,ZHANG Y L,YU L,et al.Epididymal small non-coding RNA studies:progress over the past decade[J].Andrology, 2019, 7(5):681-689.
[9]	ZHAO W S,AHMED S,LIU J X,et al.Comparative iTRAQ proteomics identified proteins associated with sperm maturation between Yak and Cattleyak epididymis[J].BMC Vet Res,2021,17(1):255.
[10]	NOWICKA-BAUER K,KURPISZ M.Current knowledge of the human sperm proteome[J].Exp Rev Proteomics,2013,10(6):591-605.
[11]	WEBER A,ARGENTI L E,DE SOUZA A P B,et al.Ready for the journey:A comparative proteome profiling of porcine cauda epididymal fluid and spermatozoa[J].Cell Tissue Res,2020,379(2):389-405.
[12]	ZHANG P F,HUANG Y L,FU Q,et al.Integrated analysis of phosphoproteome and ubiquitylome in epididymal sperm of buffalo (Bubalus bubalis)[J].Mol Reprod Dev,2021,88(1):15-33.
[13]	ROWLISON T,CLELAND T P,OTTINGER M A,et al.Novel proteomic profiling of epididymal extracellular vesicles in the domestic Cat reveals proteins related to sequential sperm maturation with differences observed between normospermic and teratospermic individuals[J].Mol Cell Proteomics,2020,19(12):2090-2104.
[14]	SUN Z L,LI S J,YU Y X,et al.Alterations in epididymal proteomics and antioxidant activity of mice exposed to fluoride[J].Arch Toxicol,2018,92(1):169-180.
[15]	HAMPOELZ B,ANDRES-PONS A,KASTRITIS P,et al.Structure and assembly of the nuclear pore complex[J].Annu Rev Biophys,2019,48:515-536.
[16]	SAKUMA S,D'ANGELO M A.The roles of the nuclear pore complex in cellular dysfunction,aging and disease[J].Semin Cell Dev Biol, 2017,68:72-84.
[17]	PACI G,CARIA J,LEMKE E A.Cargo transport through the nuclear pore complex at a glance[J].J Cell Sci,2021,134(2):jcs247874.
[18]	SKERRETT-BYRNE D A,ANDERSON A L,HULSE L,et al.Proteomic analysis of koala (phascolarctos cinereus) spermatozoa and prostatic bodies[J].Proteomics,2021,21(19):2100067.
[19]	KANEHISA M,SATO Y,KAWASHIMA M.KEGG mapping tools for uncovering hidden features in biological data[J].Protein Sci,2022,31(1):47-53.
[20]	SCHROEDER A B,DOBSON E T A,RUEDEN C T,et al.The ImageJ ecosystem:Open-source software for image visualization,processing,and analysis[J].Protein Sci,2021,30(1):234-249.
[21]	TANG B H,PAN Z X,YIN K,et al.Recent advances of deep learning in bioinformatics and computational biology[J].Front Genet,2019,10:214.
[22]	PING X Y,CHENG Y L,BAO J,et al.KPNA4 is involved in cataract formation via the nuclear import of p53[J].Gene,2021,786:145621.
[23]	ITMAN C,MIYAMOTO Y,YOUNG J,et al.Nucleocytoplasmic transport as a driver of mammalian gametogenesis[J].Semin Cell Dev Biol,2009, 20(5):607-619.
[24]	ALBER F,DOKUDOVSKAYA S,VEENHOFF L M,et al.The molecular architecture of the nuclear pore complex[J].Nature, 2007,450(7170):695-701.
[25]	WÄLDE S,KEHLENBACH R H.The Part and the Whole:functions of nucleoporins in nucleocytoplasmic transport[J].Trends Cell Biol,2010,20(8):461-469.
[26]	NEMERGUT M E,MIZZEN C A,STUKENBERG T,et al.Chromatin docking and exchange activity enhancement of RCC1 by histones H2A and H2B[J].Science,2001,292(5521):1540-1543.
[27]	KALITA J,KAPINOS L E,LIM R Y H.On the asymmetric partitioning of nucleocytoplasmic transport-recent insights and open questions[J].J Cell Sci,2021,134(7):jcs240382.
[28]	ROBBINS J,DILWORTHT S M,LASKEY R A,et al.Two interdependent basic domains in nucleoplasmin nuclear targeting sequence:identification of a class of bipartite nuclear targeting sequence[J].Cell,1991,64(3):615-623.
[29]	LINDSAY M E,PLAFKER K,SMITH A E,et al.Npap60/Nup50 is a tri-stable switch that stimulates importin-α:β-mediated nuclear protein import[J].Cell,2002,110(3):349-360.
[30]	GILCHRIST D,MYKYTKA B,REXACH M.Accelerating the rate of disassembly of karyopherin cargo complexes[J].J Biol Chem,2002,277(20):18161-18172.
[31]	MAJOR A T,WHILEY P A F,LOVELAND K L.Expression of nucleocytoplasmic transport machinery:clues to regulation of spermatogenic development[J].Biochim Biophys Acta Biomembr,2011,1813(9):1668-1688.
[32]	HU J J,WANG F C,YUAN Y,et al.Novel importin-α family member Kpna7 is required for normal fertility and fecundity in the mouse[J].J Biol Chem,2010,285(43):33113-33122.
[33]	MIYAMOTO Y,BOAG P R,HIME G R,et al.Regulated nucleocytoplasmic transport during gametogenesis[J].Biochim Biophys Acta Gene Regul Mech,2012,1819(6):616-630.
[34]	MIYAMOTO Y,SASAKI M,MIYATA H,et al.Genetic loss of importin α4 causes abnormal sperm morphology and impacts on male fertility in mouse[J].FASEB J,2020,34(12):16224-16242.
[35]	YOUNG J C,LY-HUYNH J D,LESCESEN H,et al.The nuclear import factor importin α4 can protect against oxidative stress[J].Biochim Biophys Acta Mol Cell Res,2013,1833(10):2348-2356.

Study on Nuclear-plasma Transporter KPNA4 of Sheep Sperm Cell

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 35

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WANG Haibo, ZHAN Jinshun, GU Zhiyong, CHEN Xinfeng, PAN Yue, JIA Haobin, ZHONG Xiaojun, LI Kairong, ZHAO Shengguo, HUO Junhong. Comparative Study on Meat Quality Characteristics of Three-Way Hybrid Sheep Charolais×Duper×Hu and Charolais×Australian White×Hu and Hu Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 110-119.
[2]	DUAN Xiangru, KANG Jia, YANG Ruochen, SHAN Xinyu, LI Taichun, ZHAO Wen, ZHANG Yingjie, LIU Yueqin. Effect of L-cysteine on Proliferation, Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 179-191.
[3]	YAO Ying, ZHOU Yingcong, DU Peiyan, LI Yijuan, QIAN Wenjie, LI Liuyang, YU Zhipeng, CUI Yan, YU Sijiu, FAN Jiangfeng. Proteomic Analysis of Yak Serum During Pregnancy Based on TMT Technology [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 192-206.
[4]	ZHANG Hanyue, ZHAO Dan, LIANG Yu, ZHAO Bishi, FAN Mengdan, QIAO Liying, LIU Jianhua, YANG Kaijie, PAN Yangyang, LIU Wenzhong. miR-150 Regulates Ovine Preadipocyte Differentiation by Targeting AOC3 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3262-3274.
[5]	LIU Linli, PENG Xuelan, LI Bo, CHENG Lefan, CIREN Lamu, ZHANG Enping. Effect of Overexpression of UCP3 Gene on the Differentiation of Sahu Sheep Preadipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3275-3285.
[6]	HE Mingyang, MA Yujing, WANG Yong, YANG Ruochen, LIU Yueqin, ZHANG Yingjie, DUAN Chunhui. Effects of Melatonin on Proliferation, Apoptosis of Ovarian Granulosa Cells, and Its Secretion of Steroid Hormones of Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3313-3324.
[7]	SONG Meijun, HAO Kexing, HAI Siyu, CHEN Yan, WANG Jing, HU Guangdong. Effects of SRIF-14 on Proliferation and Apoptosis of Endometrial Epithelial Cells in Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3325-3334.
[8]	LI Yuexin, LIU Aiju, MA Xiaofei, ZHENG Zhong, HU Boxin, ZHI Yunxia, TIAN Shujun. Effect of TGFβR1 on the Function of Sheep Granulosa Cells Mediated by TGF-β/Smad Signaling Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3335-3347.
[9]	MA Youji, CHEN Pengfei, MA Qing, WU Yi. Effects of Different Exercise Amounts on Rumen Microflora Diversity of Tan Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3393-3405.
[10]	LIU Xinhuan, YUN Jialei, MAO Li, LI Jizong, HAO Fei, HE Miaofeng, YANG Leilei, ZHANG Wenwen, CHENG Zilong, SUN Min, LIU Maojun, WANG Shaohui, BAI Juan, LI Wenliang. Isolation, Identification, Virulence Genes and Drug Resistance Analysis of Escherichia coli Isolated from Diarrheal Goat and Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3445-3454.
[11]	ZHANG Ying, JIN Hua, HAN Yang, SUI Dan, XIAO Xin, HAO Xiujing, LI Min. Analysis of Microbial Diversity in Nasal Cavity of Sheep with Respiratory Disease and Their Environment [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3455-3465.
[12]	XIONG Chengkun, ZHANG Daoliang, YANG Yue, DING Hongyan, ZHAO Jie, LI Yu, WANG Xichun, FENG Shibin, ZHAO Chang, TANG Jishun, WU Jinjie. Effect of Rutin on Rumen Fermentation, Rumen Flora Structure and Antioxidant Properties in Perinatal Hu Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2898-2909.
[13]	DONG Yajie, HAO Xiaojing, WU Jinqiang, WANG Rong, ZHANG Pengxiang, WANG Haidong, HE Xiaoyan. Exploration of the Effect of SHH on Wool Bending through Krox20 Regulation of IGFBP5 Expression Based on Sheep Keratinocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2365-2375.
[14]	ZHAO Donghao, YUAN Meng, MA Kaiteng, DUAN Zhuo, ZHU Yixin, TANG Fang, HAN Keguang, HUO Nairui. Chelating Role of Sheep Bone Collagen Peptide to Cadmium and Its Protection Role against Liver Injuries Induced by Cadmium in Chickens [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2641-2652.
[15]	MA Keyan, HAN Jintao, BAI Yaqin, LI Taotao, MA Youji. Genetic Diversity Analysis of Yongdeng Qishan Sheep Based on Specific-Locus Amplified Fragment Sequencing [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 1939-1950.