水牛水通道蛋白9基因克隆及其表达分析

doi:10.16431/j.cnki.1671-7236.2015.07.001

Abstract

Abstract: Cloning buffalo AQP9 gene and analyzing its expression in buffalo tissues.A pair of primers was designed according to the released bovine AQP9 sequences in GenBank,which was used to clone buffalo AQP9 gene.The AQP9 gene was amplified by RT-PCR,whose nucleotide sequence and protein structure were analyzed by bioinformatics methods.The expression of AQP9 in buffalo tissues was assayed by Real-time quantitative PCR.The expression of AQP9 gene in buffalo ovary and testis tissue was detected by immunohistochemical staining method.The results showed that the cloned ORF length of buffalo AQP9 gene was 888 bp,which coded 295 amino acids.The results of multiple sequence comparison showed that the nucleotide sequence of buffalo AQP9 shared 99%,90%,97% and 88% homologeous compared with that of Bos taurus,Sus scrofa,Ovis ariessis and Homo sapiens,respectively,while shared 99%,86%,97%,83% homologeous for amino acids,respectively.Phylogenetic tree analysis indicated that AQP9 gene was highly conservative in the evolutionary process.Real-time quantitative PCR results showed that AQP9 gene expressed in buffalo liver,lung,brain,skin,testis and ovary tissues with different levels,had the most abundant expression in liver,followed by in skin and testis,less observed in lung and ovary.The results of immunohistochemical staining showed that the expression of AQP9 protein varied with the development of buffalo ovarian tissue,and gradually enhanced with follicle development.In testicular tissue,AQP9 protein expressed in spermatocyte and leydig cells of developmental stage testis.These results indicated that we had successfully cloned buffalo AQP9 gene sequences.The expression and its function of AQP9 in buffalo ovaries and testes might play an important role in follicle development and spermatogenesis.

Key words: buffalo; aquaporins; AQP9 gene; cloning; expression analysis

CLC Number:

QU Chun-feng, LI Sheng, LI Hui, DU Feng-jiao, LEI Wei, WU Zhu-lian, LI Xiang-ping, SHI De-shun. Cloning of Buffalo AQP9 Gene and Analysing its Expression in the Buffalo Tissues[J]. , 2015, 42(7): 1621-1629.

References

[1] Preston G M,Carroll T P,Guggino W B,et al.Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein[J]. Science,1992,256(5055):385-387.
[2] Huang H F,He R H,Sun C C,et al.Function of aquaporins in female and male reproductive systems[J].Human Reproduction Update,2006,12(6):785-795.
[3] Zhang D,Tan Y J,Qu F,et al.Functions of water channels in male and female reproductive systems[J].Mol Aspects Med,2012,33(5-6):676-690.
[4] McConnell N A,Yunus R S,Gross S A,et al.Water permeability of an ovarian antral follicle is predominantly transcellular and mediated by aquaporins[J].Endocrinology,2002,143(8):2905-2912.
[5] 刘絮,张忠芳,赵丹,等.不同发育阶段大鼠睾丸水通道蛋白AQP9的表达研究[J].北华大学学报(自然科学版),2004,6:509-511.
[6] Jablonski E M.Plasma membrane aquaporin activity can affect the rate of apoptosis but is inhibited after apoptotic volume decrease[J].Am J Physiol Cell Physiology,2004,286(4):975C-985C.
[7] Kobayashi K,Yasui M.Cellular and subcellular localization of aquaporins 1,3,8,and 9 in amniotic membranes during pregnancy in mice[J].Cell and Tissue Research,2010,342(2):307-316.
[8] Yeung C H,Callies C,Tüttelmann F,et al.Aquaporins in the human testis and spermatozoa-identification,involvement in sperm volume regulation and clinical relevance[J].International Journal of Andrology,2009,33:629-641.
[9] Tsukaguchi H,Shayakul C,Berger U V,et al.Molecular characterization of a broad selectivity neutral solute channel[J]. J Biol Chem,1998,273(38):24737-24743.
[10] Ishibashi K,Kuwahara M,Gu Y,et al.Cloning and functional expression of a new aquaporin (AQP9) abundantly expressed in the peripheral leukocytes permeable to water and urea,but not to glycerol[J].Biochem Biophys Res Commun,1998,244(1):268-274.
[11] Ko S B,Uchida S,Naruse S,et al.Cloning and functional expression of rAOP9L a new member of aquaporin family from rat liver[J].Biochem Mol Biol Int,1999,47(2):309-318.
[12] Wang S,Chen J,Huang B,et al.Cloning and cellular expression of aquaporin 9 in ovine fetal membranes[J].American Journal of Obstetrics and Gynecology,2005,193(3):841-848.
[13] 李晓峰.猪水-甘油通道家族基因的克隆、染色体定位及组织表达谱[D].武汉:华中农业大学,2008.
[14] Kuriyama H,Kawamoto S,Ishida N,et al.Molecular cloning and expression of a novel human aquaporin from adipose tissue with glycerol permeability[J].Biochem Biophys Res Commun,1997,241(1):53-58.
[15] Tsukaguchi H,Weremowicz S,Morton C C,et al.Functional and molecular characterization of the human neutral solute channel aquaporin-9[J].Am J Physiol,1999,277(2):F685-F696.
[16] 张乐颖,王加启,卜登攀,等.水通道蛋白研究进展[J].中国畜牧兽医,2008,35(11):40-43.
[17] Pastor-Soler N,Isnard-Bagnis C,Herak-Kramberger C,et al.Expression of aquaporin 9 in the adult rat epididymal epithelium is modulated by androgens[J].Biol Reprod,2002,66(6):1716-1722.
[18] Elkjaer M L,Vajda Z,Nejsum L N,et al.Immunolocalization of AQP9 in liver,epididymis,testis,spleen,and brain[J]. Biochemical and Biophysical Research Communications,2000,276(3):1118-1128.
[19] 杨安素,邹双微,华莹,等.子痫前期产妇胎盘胎膜中水通道蛋白8和水通道蛋白9的表达和意义[J].医学研究杂志,2012,41(11):83-86.
[20] Yang J H,Yan C X,Chen X J,et al.Expression of aquaglyceroporins in epithelial ovarian tumours and their clinical significance[J]. J Int Med Res,2011,39(3):702-711.
[21] Saito K,Kageyama Y,Okada Y,et al.Localization of aquaporin-7 in human testis and ejaculated sperm:Possible involvement in maintenance of sperm quality[J].J Urol,2004,172(1):2073-2076.
[22] Sun X,Zhang J,Fan Y,et al.Aquaporin-4 deficiency induces subfertility in female mice[J].Fertility and Sterility,2009,92(5):1736-1743.
[23] Sha X,Xiong Z,Liu H,et al.Pregnant phenotype in aquaporin 8-deficient mice[J].Acta Pharmacologica Sinica,2011,32(6):840-844.
[24] Pastor-Soler N M,Fisher J S,Sharpe R,et al.Aquaporin 9 expression in the developing rat epididymis is modulated by steroid hormones[J].Reproduction,2010,139(3):613-621.
[25] Wang J,Tanji N,Sasaki T,et al.Androgens upregulate aquaporin 9 expression in the prostate[J].International Journal of Urology,2008,15(10):936-941.
[26] Qu F,Wang F F,Lu X E,et al.Altered aquaporin expression in women with polycystic ovary syndrome:Hyperandrogenism in follicular fluid inhibits aquaporin-9 in granulosa cells through the phosphatidylinositol 3-kinase pathway[J].Human Reproduction,2010,25(6):1441-1450.
[27] 李胜,屈春凤,李润,等.水牛水通道蛋白8基因克隆及表达分析[J].中国畜牧兽医,2014,41(3):44-48.

Cloning of Buffalo AQP9 Gene and Analysing its Expression in the Buffalo Tissues

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