绵羊INSL3基因组织表达规律与潜在功能位点分析研究

doi:10.11843/j.issn.0366-6964.2020.04.004

Abstract

Abstract: The aim of this study was to analyze the tissue expression pattern and functional SNP of INSL3 gene in sheep. In this study, three each of male and female 1-year-old Small Tail Han sheep with healthy body condition and normal horn were used to detect tissue expression pattern of INSL3 by RT-PCR and real-time PCR. The gene resequencing data of 10 sheep breeds were used to explore the principal component analysis (PCA) and SNP of INSL3 gene region. The qPCR result showed that the expression level of INSL3 gene was the highest in ovary and testis, and INSL3 gene was also expressed in soft horn. The expression of INSL3 gene in testis was significantly higher than that in ovary (P<0.01), and the expression of INSL3 gene in soft horn of ram was significantly higher than that in ewe (P<0.05). PCA results showed that the INSL3 gene region was clustered according to the presence or absence of horn in a certain extent, which further indicated that this region might be related to horn. Functional site analysis revealed that 4 potential SNP sites were significantly different in distribution in horned and hornless populations. SNP1 was at the transcription factor binding site of KDM1A, SNP3 at the ESR1 binding site, and SNP6 at the terminator region. A missense mutation and a synonymous mutation were also found. This study shows that INSL3 gene may be related to the different horn size between male and female sheep as well as presence or absence of the horn, and multiple potential functional sites have been found. This study provides the basis for the future research on the functional mechanism of INSL3.

Key words: INSL3, real-time PCR, tissue expression pattern, functional SNP, sheep

CLC Number:

S826.2

PAN Zhangyuan, ZHANG Zijie, JI Jiuxiu, LI Huazhen, SUN Anfang, LI Fukuan, WANG Hui, CHU Mingxing, Lü Shenjin. Tissue Expression Pattern and Functional SNP Analysis of INSL3 Gene in Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 679-687.

References 43

[1]	国家畜禽遗传资源委员会.中国畜禽遗传资源志-羊志[M].北京:中国农业出版社,2011:1-16.National Commission of Livestock and Poultry Genetic Resources.Animal genetic resources in China:sheep and goats[M].Beijing:China Agriculture Press,2011:1-16.(in Chinese)
[2]	NEF S,PARADA L F.Cryptorchidism in mice mutant for Insl3[J].Nat Genet,1999,22(3):295-299.
[3]	FENG S,FERLIN A,TRUONG A,et al.INSL3/RXFP2 signaling in testicular descent:mice and men[J].Ann N Y Acad Sci,2009, 1160(1):197-204.
[4]	IVELL R,ANAND-IVELL R.Insulin-like peptide 3(INSL3) is a major regulator of female reproductive physiology[J].Hum Reprod Update,2018,24(6):639-651.
[5]	FERLIN A,DE TONI L,AGOULNIK A I,et al.Protective role of testicular hormone INSL3 from atrophy and weakness in skeletal muscle[J].Front Endocrinol,2018,9:562.
[6]	IVELL R,BATHGATE R A D.Reproductive biology of the relaxin-like factor (RLF/INSL3)[J].Biol Reprod,2002, 67(3):699-705.
[7]	HAMPEL U,KLONISCH T,SEL S,et al.Insulin-like factor 3 promotes wound healing at the ocular surface[J].Endocrinology,2013,154(6):2034-2045.
[8]	IVELL R,ANAND-IVELL R.Biology of Insulin-like Factor 3 in human reproduction[J].Hum Reprod Update, 2009,15(4):463-476.
[9]	FERLIN A,GAROLLA A,RIGON F,et al.Changes in serum insulin-like factor 3 during normal male puberty[J].J Clin Endocrinol Metab,2006,91(9):3426-3431.
[10]	赵丽丽,聂敏,伍学焱.胰岛素样肽3的功能及其研究进展[J].生殖医学杂志,2016,25(4):374-377.ZHAO L L,NIE M,WU X Y.Function insulin-like peptide 3 and its research progress[J].Journal of Reproductive Medicine,2016,25(4):374-377.(in Chinese)
	BAY K,COHEN A S,JØRGENSEN F S,et al.Insulin-like factor 3 levels in second-trimester amniotic fluid[J].J Clin Endocrinol Metab,2008,93(10):4048-4051.
[11]	HARRISON S M,BUSH N C,WANG Y,et al.Insulin-Like Peptide 3(INSL3) serum concentration during human male fetal life[J].Front Endocrinol,2019,10:596.
[12]	BATHGATE R,BALVERS M,HUNT N,et al.Relaxin-like factor gene is highly expressed in the bovine ovary of the cycle and pregnancy:sequence and messenger ribonucleic acid analysis[J].Biol Reprod,1996,55(6):1452-1457.
[13]	BATHGATE R,MONIAC N,BARTLICK B,et al.Expression and regulation of relaxin-like factor gene transcripts in the bovine ovary:differentiation-dependent expression in theca cell cultures[J].Biol Reprod,1999,61(4):1090-1098.
[14]	ANAND-IVELL R,HIENDLEDER S,VIÑOLES C,et al.INSL3 in the ruminant:a powerful indicator of gender- and genetic-specific feto-maternal dialogue[J].PLoS One,2011,6(5):e19821.
[15]	ANAND-IVELL R, HIENDLEDER S, VIÑOLES C, et al. INSL3 in the ruminant:a powerful indicator of gender- and genetic-specific feto-maternal dialogue[J]. PLoS One, 2011,6(5):e19821.
[16]	DOMINIK S,HENSHALL J M,HAYES B J.A single nucleotide polymorphism on chromosome 10 is highly predictive for the polled phenotype in Australian Merino sheep[J].Anim Genet,2012,43(4):468-470.
[17]	JOHNSTON S E,MCEWAN J C,PICKERING N K,et al.Genome-wide association mapping identifies the genetic basis of discrete and quantitative variation in sexual weaponry in a wild sheep population[J].Mol Ecol,2011,20(12):2555-2566.
[18]	PAN Z Y,LI S D,LIU Q Y,et al.Whole-genome sequences of 89 Chinese sheep suggest role of RXFP2 in the development of unique horn phenotype as response to semi-feralization[J].Gigascience,2018,7(4):giy019.
[19]	DOVE W F.The physiology of horn growth:a study of the morphogenesis,the interaction of tissues,and the evolutionary processes of a mendelian recessive character by means of transplantation of tissues[J].J Exp Zool,1935,69(3):347-405.
[20]	FERLIN A,PERILLI L,GIANESELLO L,et al.Profiling insulin like factor 3(INSL3) signaling in human osteoblasts[J].PLoS One,2011,6(12):e29733.
[21]	FERLIN A,SELICE R,CARRARO U,et al.Testicular function and bone metabolism-beyond testosterone[J].Nat Rev Endocrinol,2013,9(9):548-554.
[22]	FERLIN A,SELICE R,DI MAMBRO A,et al.Role of vitamin D levels and vitamin D supplementation on bone mineral density in Klinefelter syndrome[J].Osteoporos Int,2015,26(8):2193-2202.
[23]	FERLIN A,DE TONI L,SANDRI M,et al.Relaxin and insulin-like peptide 3 in the musculoskeletal system:from bench to bedside[J].Brit J Pharmacol,2017,174(10):1015-1024.
[24]	DE TONI L,AGOULNIK A I,SANDRI M,et al.INSL3 in the muscolo-skeletal system[J].Mol Cell Endocrinol, 2019, 487:12-17.
[25]	DI NISIO A,DE TONI L,ROCCA M S,et al.Negative association between sclerostin and INSL3 in isolated human osteocytes and in klinefelter syndrome:new hints for testis-bone crosstalk[J].J Clin Endocrinol Metab, 2018, 103(5):2033-2041.
[26]	潘章源,贺小云,刘秋月,等.绵羊GDF9基因mRNA、DNA和调控区序列克隆及其在11个品种中遗传多态性检测[J].畜牧兽医学报,2016,47(8):1555-1564.PAN Z Y,HE X Y,LIU Q Y,et al.Cloning and genetic polymorphism analysis of mRNA,DNA and regulatory region of ovine GDF9 gene in 11 breeds[J].Acta Veterinaria et Zootechnica Sinica,2016,47(8):1555-1564.(in Chinese)
[27]	卢璐璐,孙晓笛,储明星,等.绵羊AA-NAT基因mRNA表达与常年发情相关性研究[J].畜牧兽医学报,2015,46(4):542-548.LU L L,SUN X D,CHU M X,et al.Association between AA-NAT mRNA expression and year-round estrus in sheep[J].Acta Veterinaria et Zootechnica Sinica,2015,46(4):542-548.(in Chinese)
[28]	潘章源,贺小云,刘秋月,等.全基因组测序(WGS)在畜禽群体进化和功能基因挖掘中的应用[J].农业生物技术学报,2016,24(12):1945-1954.PAN Z Y,HE X Y,LIU Q Y,et al.Application of whole genome sequencing (WGS) in population evolution and gene discovery of domestic animals[J].Journal of Agricultural Biotechnology,2016,24(12):1945-1954.(in Chinese)
[29]	KAWAJI H,LIZIO M,ITOH M,et al.Comparison of CAGE and RNA-seq transcriptome profiling using clonally amplified and single-molecule next-generation sequencing[J].Genome Res,2014,24(4):708-717.
[30]	MERKIN J,RUSSELL C,CHEN P,et al.Evolutionary dynamics of gene and isoform regulation in mammalian tissues[J].Science,2012,338(6114):1593-1599.
[31]	HUNTLEY M A,LOU M,GOLDSTEIN L D,et al.Complex regulation of ADAR-mediated RNA-editing across tissues[J].BMC Genomics,2016,17(1):61.
[32]	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,2014,5:3230.
[33]	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.
[34]	PAN Z Y,LI S D,LIU Q Y,et al.Rapid evolution of a retro-transposable hotspot of ovine genome underlies the alteration of BMP2 expression and development of fat tails[J].BMC Genomics,2019,20(1):261.
	HU H,MIAO Y R,JIA L H,et al.AnimalTFDB 3.0:a comprehensive resource for annotation and prediction of animal transcription factors[J].Nucleic Acids Res,2019,47(D1):D33-D38.
[36]	SPANEL-BOROWSKI K,SCHÄFER I,ZIMMERMANN S,et al.Increase in final stages of follicular atresia and premature decay of corpora lutea in Insl3-deficient mice[J].Mol Reprod Dev,2001,58(3):281-286.
[37]	JOHNSTON S E,GRATTEN J,BERENOS C,et al.Life history trade-offs at a single locus maintain sexually selected genetic variation[J].Nature,2013,502(7469):93-95.
[38]	LÜHKEN G,KREBS S,ROTHAMMER S,et al.The 1.78-kb insertion in the 3'-untranslated region of RXFP2 does not segregate with horn status in sheep breeds with variable horn status[J].Genet Sel Evol,2016,48(1):78.
[39]	SHI Y J,LAN F,MATSON C,et al.Histone demethylation mediated by the nuclear amine oxidase homolog LSD1[J]. Cell,2004,119(7):941-953.
[40]	PEDERSEN M T,HELIN K.Histone demethylases in development and disease[J].Trends Cell Biol,2010,20(11):662-671.
[41]	DAHLMAN-WRIGHT K,CAVAILLES V,FUQUA S A,et al.International union of pharmacology.LXIV.Estrogen receptors[J]. Pharmacol Rev,2006,58(4):773-781.
[42]	DI PAOLA R,FRITTITTA L,MISCIO G,et al.A variation in 3' UTR of hPTP1B increases specific gene expression and associates with insulin resistance[J].Am J Hum Genet,2002,70(3):806-812.
[43]	NOSSENT A Y,HANSEN J L,DOGGEN C,et al.SNPs in microRNA binding sites in 3'-UTRs of RAAS genes influence arterial blood pressure and risk of myocardial infarction[J].Am J Hypertens,2011,24(9):999-1006.

Tissue Expression Pattern and Functional SNP Analysis of INSL3 Gene in Sheep

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