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

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