犏牛PPP1R11基因的克隆及在睾丸中的表达规律研究

doi:10.11843/j.issn.0366-6964.2022.02.015

Abstract

Abstract: This study was conducted to clone the PPP1R11 gene in cattle-yak, detect the expression and location in testes of cattle-yak at different developmental stages, as to provide a theoretical basis for further research of the mechanism of function in male reproduction. The testis, epididymis, heart, liver, spleen, lung, kidney, large intestine, small intestine, stomach, muscle and adipose tissue of adult cattle-yak were collected (n=3), testicles of cattle-yak and yak in the fetal period (5-6 months old), juvenile period (1-2 years old) and adult period (3-4 years old) (n=3) were collected as experimental materials. The PPP1R11 CDS region of cattle-yak was cloned by RT-PCR and analyzed by bioinformatics softwares. qRT-PCR was used to detect the expression patterns of PPP1R11 mRNA in different tissues of cattle-yak, and compared the expression difference in testes at different development stages between cattle-yak and yak. The cell location and expression of PPP1R11 protein were detected by immunohistochemistry (IHC) staining. The results showed that the CDS region of PPP1R11 gene in cattle-yak was 324 bp, which encoded 107 amino acids. The corresponding PPP1R11 protein of cattle-yak had high homology with other mammals. The protein function prediction showed that PPP1R11 could interact with PPP1R2, PPP1R7, PPP1CB, UTP20 and other proteins, and mainly related to the phosphorylation of proteins, which could regulate biological processes such as testicular development, spermatogenesis and sexual maturity. PPP1R11 gene was wildly expressed in various tissues of cattle-yak, and the relative expression level in testis was significantly higher than other tissues (P<0.01). The expression of PPP1R11 gene in cattle-yak testis increased with age. Furthermore, there was significant difference of the expression level of PPP1R11 (P<0.01) in testis at juvenile and adult stage of cattle-yak compared with the counterpart of yak. In addition, IHC staining result showed that PPP1R11 protein was significantly lower expressed in cattle-yak spermatogonia and sertoli cells, and there was significant difference compared with yak, and the number of primary spermatocytes in cattle-yak was reduced significantly and meiosis arrested at this stage. This study showed that there were temporal and spatial differences in the expression of PPP1R11 between cattle-yak and yak testis, suggesting that PPP1R11 may be related to to male cattle-yak infertility. However, its specific mechanism needs to be further studied.

Key words: cattle-yak, PPP1R11, gene cloning, gene expression, meiosis, testis development

CLC Number:

S823.85

MIN Xingyu, YANG Lixue, YU Hailing, HU Yulei, YANG Manzhen, YANG Luyu, LI Jian, XIONG Xianrong. Cloning and Expression Analysis of Cattle-yak PPP1R11 Gene in Testis[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 481-492.

References 36

[1]	XU C F,SHAH M A,MIPAM T,et al.Bovid microRNAs involved in the process of spermatogonia differentiation into spermatocytes[J].Int J Biol Sci,2020,16(2):239-250.
[2]	WU Y,ZHANG W X,ZUO F,et al.Comparison of mRNA expression from Y-chromosome X-degenerate region genes in taurine cattle,yaks and interspecific hybrid bulls[J].Anim Genet,2019,50(6):740-743.
[3]	SONG C C,HUANG Y Z,YANG Z X,et al.RNA-Seq analysis identifies differentially expressed genes insubcutaneous adipose tissuein Qaidamford cattle,cattle-yak,and angus cattle[J].Animals (Basel),2019,9(12):1077.
[4]	NIAYALE R,CUI Y,ADZITEY F.Male hybrid sterility in the cattle-yak and other bovines:a review[J].Biol Reprod,2021,104(3):495-507.
[5]	阿果约达,熊显荣,王艳,等.犏牛雄性不育相关lncRNA的鉴定与分析[J].畜牧兽医学报,2019,50(3):551-561.A GUO Y D,XIONG X R,WANG Y,et al.Identification and analysis of long non-coding RNA associated with cattle-yak male infertility[J].Acta Veterinaria et Zootechnica Sinica,2019,50(3):551-561.(in Chinese)
[6]	BOLLEN M,PETI W,RAGUSA M J,et al.The extended PP1 toolkit:designed to create specificity[J].Trends Biochem Sci, 2010,35(8):450-458.
[7]	UBERSAX J A,FERRELL JR J E.Mechanisms of specificity in protein phosphorylation[J].Nat Rev Mol Cell Biol,2007,8(7):530-541.
[8]	DUDIKI T,JOUDEH N,SINHA N,et al.The protein phosphatase isoform PP1γ1 substitutes for PP1γ2 to support spermatogenesis but not normal sperm function and fertility[J].Biol Reprod,2019,100(3):721-736.
[9]	DEY S,BROTHAG C,VIJAYARAGHAVAN S.Signaling enzymes required for sperm maturation and fertilization in mammals[J]. Front Cell Dev Biol,2019,7:341.
[10]	CHAKRABARTI R,CHENG L N,PURI P,et al.Protein phosphatase PP1γ2 in sperm morphogenesis and epididymal initiation of sperm motility[J].Asian J Androl,2007,9(4):445-452.
[11]	CHAKRABARTI R,KLINE D,LU J,et al.Analysis of Ppp1cc-null mice suggests a role for PP1 gamma2 in sperm morphogenesis[J]. Biol Reprod,2007,76(6):992-1001.
[12]	LI H H,ROKAVEC M,JIANG L C,et al.Antagonistic effects of p53 and HIF1A on microRNA-34a regulation of PPP1R11 and STAT3 and hypoxia-induced epithelial to mesenchymal transition in colorectal cancer cells[J].Gastroenterology, 2017, 153(2):505-520.
[13]	CHENG L N,PILDER S,NAIRN A C,et al.PP1γ2 and PPP1R11 are parts of a multimeric complex in developing testicular germ cells in which their steady state levels are reciprocally related[J].PLoS One,2009,4(3):e4861.
[14]	HAN Y B,FENG H L,CHEUNG C K,et al.Expression of a novel T-complex testis expressed 5(Tctex5) in mouse testis, epididymis,and spermatozoa[J].Mol Reprod Dev,2007,74(9):1132-1140.
[15]	GOSWAMI S,KORRODI-GREGÓRIO L,SINHA N,et al.Regulators of the protein phosphatase PP1γ2,PPP1R2,PPP1R7,and PPP1R11 are involved in epididymal sperm maturation[J].J Cell Physiol,2019,234(3):3105-3118.
[16]	黄向月,熊显荣,张磊,等.SPATA3基因克隆及其在牦牛和犏牛睾丸中的差异表达研究[J].畜牧兽医学报, 2020,51(2):279-287.HUANG X Y,XIONG X R,ZHANG L,et al.Cloning of SPATA3 gene and its differential expression analysis in yak and cattle-yak testis[J].Acta Veterinaria et Zootechnica Sinica,2019,51(2):279-287.(in Chinese)
[17]	ZHANG J Y,MRUK D D,CHENG C Y.Myotubularin phosphoinositide phosphatases,protein phosphatases,and kinases:their roles in junction dynamics and spermatogenesis[J].J Cell Physiol,2005,204(2):470-483.
[18]	FORGIONE N,VOGL A W,VARMUZA S.Loss of protein phosphatase 1cγ (PPP1CC) leads to impaired spermatogenesis associated with defects in chromatin condensation and acrosome development:an ultrastructural analysis[J].Reproduction, 2010, 139(6):1021-1029.
[19]	ZIMIN A V,DELCHER A L,FLOREA L,et al.A whole-genome assembly of the domestic cow,Bos taurus[J].Genome Biol,2009, 10(4):R42.
[20]	DUDIKI T,KADUNGANATTIL S,FERRARA J K,et al.Changes in carboxy methylation and tyrosine phosphorylation of protein phosphatase PP2A are associated with epididymal sperm maturation and motility[J].PLoS One,2015,10(11):e0141961.
[21]	VAN DEN HAM R,VAN DISSEL-EMILIANI F M F,VAN PELT A M M.Expression of the scaffolding subunit A of protein phosphatase 2A during rat testicular development[J].Biol Reprod,2003,68(4):1369-1375.
[22]	SILVA J V,FREITAS M J,FARDILHA M.Phosphoprotein phosphatase 1 complexes in spermatogenesis[J].Curr Mol Pharmacol, 2014,7(2):136-146.
[23]	MACLEOD G,VARMUZA S.Tandem affinity purification in transgenic mouse embryonic stem cells identifies DDOST as a novel PPP1CC2 interacting protein[J].Biochemistry,2012,51(48):9678-9688.
[24]	PAN X Y,CHEN X,TONG X,et al.Ppp2ca knockout in mice spermatogenesis[J].Reproduction,2015,149(4):385-391.
[25]	CAI X,YU S M,MIPAM T,et al.Comparative analysis of testis transcriptomes associated with male infertility in cattleyak[J]. Theriogenology,2017,88:28-42.
[26]	KORNBLUTH S,FISSORE R.Vertebrate reproduction[J].Cold Spring Harb Perspect Biol,2015,7(10):a006064.
[27]	EITENEUER A,SEILER J,WEITH M,et al.Inhibitor-3 ensures bipolar mitotic spindle attachment by limiting association of SDS22 with kinetochore-bound protein phosphatase-1[J].EMBO J,2014,33(22):2704-2720.
[28]	PENG Q H,WU J G,ZHANG Y,et al.1A6/DRIM,a novel t-UTP,activates RNA polymerase I transcription and promotes cell proliferation[J].PLoS One,2010,5(12):e14244.
[29]	YUE F,CHENG Y,BRESCHI A,et al.A comparative encyclopedia of DNA elements in the mouse genome[J].Nature, 2014,515(7527):355-364.
[30]	YU Y,FUSCOE J C,ZHAO C,et al.A rat RNA-Seq transcriptomic BodyMap across 11 organs and 4 developmental stages[J].Nat Commun,2014,5(1):3230.
[31]	JIANG Y,XIE M,CHEN W B,et al.The sheep genome illuminates biology of the rumen and lipid metabolism[J]. Science,2014,344(6188):1168-1173.
[32]	LIU G S,WU J J,QIAO M,et al.Transcription elements AREB6 and miR-34a affect apoptosis of PAMs by regulating the expression of SS2-related gene PPP1R11[J].Cell Cycle,2019,18(9):1033-1044.
[33]	HAN Y B,SONG X X,FENG H L,et al.Mutations of t-complex testis expressed gene 5 transcripts in the testis of sterile t-haplotype mutant mouse[J].Asian J Androl,2008,10(2):219-226.
[34]	ZOU S,LIU P,YU S,et al.Hsp60 expression and localization in different tissues and testis development of male cattle (cattle-yak and yak)[J].Folia Morphol (Warsz),2021,doi:10.5603/FM.a2020.0126.
[35]	SATO Y,KURIWAKI R,HAGINO S,et al.Abnormal functions of Leydig cells in crossbred cattle-yak showing infertility[J]. Reprod Domest Anim,2020,55(2):209-216.
[36]	LIU P G,YU S J,CUI Y,et al.Regulation by Hsp27/P53 in testis development and sperm apoptosis of male cattle (cattle-yak and yak)[J].J Cell Physiol,2018,234(1):650-660.

Cloning and Expression Analysis of Cattle-yak PPP1R11 Gene in Testis

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 36

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	YANG Yang, YU Qian, LIU Yucheng, YANG Hua, ZHAO Zhuo, WANG Limin, ZHOU Ping, YANG Qingyong, DAI Rong. Identification and Tissue Expression Analysis of the Sheep MYL Gene Family [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1551-1564.
[2]	HUANG Xianpeng, XING Jiayi, BAI Yuanyuan, JIANG Yuting, MA Zhiwei, FU Wei, LAN Daoliang. Cloning of Six Pluripotent Related Transcription Factors OSKMNL in Yak and Construction of Polycistron Lentiviral Vector [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1579-1591.
[3]	KANG Jia, DUAN Xiangru, YIN Xuejiao, YANG Ruochen, LI Taichun, SHAN Xinyu, CHEN Meijing, ZHANG Yingjie, LIU Yueqin. Effects of Cysteine and Methionine on Secondary Hair Follicle Growth and Hair Dermal Papilla Cell Proliferation in vitro in Cashmere Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 515-527.
[4]	SHAO Yuexin, ZHANG Xinyu, GE Liyan, SHI Huaiping. Cloning and RNA Interference Analysis of ATF4 Gene in Xinong Saanen Dairy Goat [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2353-2364.
[5]	CHEN Cancan, JIANG Jing, SUN Xiaoyan, REN Hangxing, LI Jie. Expression of AGRP Gene in Goat Tissue and Its Action Mechanism on Melanin Production [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1441-1451.
[6]	SHAO Jing, ZHANG Ying, TANG Yu, XU Baozeng. Research Progress of ERM Proteins in Mammalian Oocyte Maturation and Fertilization [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4477-4487.
[7]	JI Zhengyu, NI Mengru, ZHANG Zhaobo, ZHAO Gan, HUANG Zan, LI Pinghua, HUANG Ruihua, HOU Liming. Research on Adipogenic Capacity and Gene Expression Pattern of in Vitro FAPs Cells Derived from Longissimus Dorsi Muscle of Suhuai Pig [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4126-4142.
[8]	SANG Lei, CHEN Dongjin, SUN Shikun, GAO Chengfang, WANG Jinxiang, CHEN Yanfeng, XIE Xiping. Cloning and Expression of GnIH Gene and Its Effect on Reproductive Hormones Secretion of Young Male Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 201-212.
[9]	YANG Yang, ZHOU Ziwei, ZHANG Jingyi, YANG Shuo, WANG Boyu, GE Nan, LIN Ye, HOU Xiaoming. Analysis on SP1 Gene Structure and Its Function on Milk Fat Synthesis in Holstein Dairy Cows [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 2970-2981.
[10]	LUORENG Zhuoma, WANG Jinpeng, JIAO Peng, LI Yanxia, DONG Yiwen, WEI Dawei, WNAG Xingping. Construction of Dairy Cow Mastitis Model and Analysis of mRNA Transcription Level of Inflammation Related Cytokine Genes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2763-2772.
[11]	YANG Hongzao, ZHU Jie, GUO Hongyuan, TANG Aoxing, CHEN Shaoyu, LIU Chuncao, ZHANG Da, YUAN Ligang, LIU Guangqing. Expression and Subcellular Localization Analysis of Felis catus C11orf96 [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2390-2395.
[12]	LIU Kehan, WANG Yong, LI Yanyan, WANG Chongyang, ZHU Jiangjiang, XIONG Yan, LI An, LIN Yaqiu. Effects of SRSF10 on the Differentiation of Intramuscular Preadipocytes in Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1768-1778.
[13]	YU Yongtao, MAO Yanni, ZHAO Qingmei, LI Jinrong, BAI Xiaonan, XUE Jiaqi, ZHANG Haodong. Expression Pattern Analysis of Related Genes in SWN Gene Cluster of Alternaria oxytropis Mutated by Ethyl Methylate [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1241-1251.
[14]	LI Yan, LAN Tingying, PANG Bo, YANG Xiaonong, HUANG Jian. Expression of NLRP3 Inflammasome Signal Associated Genes in Canine Mammary Tumors and Clinical Significance [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1252-1258.
[15]	LIU Yue, XUE Xianglan, LI Xiaobo, JIANG Lin, PU Yabin, HE Xiaohong, MA Yuehui, ZHAO Qianjun. Research Progress of the Relationship between Chromatin Accessibility and Animal Embryo Development [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 680-687.