Abstract:Abstract The aim of the study was to clone the CDS of retinol binding protein 4 gene (RBP4), and to determine its expression in different months old testis of sheep (Ovis aires), which might provide basis for its functional research on testis development. Sheep testis was used as material in this study. The sequence of RBP4 gene was cloned by RT-PCR, and structure of RBP4 protein was analyzed with relevant bioinformatics softwares. The mRNA and protein expression of RBP4 was determined using qRT-PCR and immunohistochemistry, respectively. The results showed that the CDS length of RBP4 gene was 606 bp, and it encoded goat (Capra hircus), cow (Bos taurus), pig (Sus scrofa), human (Homo sapiens) and mouse (Mus musculus), respectively. The sheep RBP4 gene was the closest with that of goat, followed by cow, pig, human and mouse. RBP4 mRNA expressed in all different months old and the expression was firstly decreased and then increased with month increasing. The difference between the adjacent two groups were extremely significant (P<0.01). RBP4 was mainly localized in the primary spermatocytes, spermatid, spermatozoon and seminiferous tubule. The positive reaction integrated optical density showed a trend of first increasing and then decreasing with month increasing, which was opposite to the expression of RBP4 mRNA. The study result indicated that the RBP4 gene may participate in the control of sheep testis development, which provides reference to further exploration of the influence of RBP4 in sheep reproductive process.
[1]Newcomer, M. E.; Ong, D. E.Plasma retinol binding protein: structure and function of the prototypic lipocalin. [J].Biochimica Et Biophysica Acta, 2000, 1482((1-2)):57-64
[2]Smith, C. W.; Patton, J. G.; Nadal-Ginard, B. Alternative splicing in the control of gene expression[J].Genetics, 1989, 23((23)):527-577
[3]Kanai, M.; Raz, A.; Goodman, D. S.Retinol-binding protein: the transport protein for vitamin A in human plasma[J].Journal of Clinical Investigation, 1968, 47( (9)):2025-2044
[4]Gursoy, A. Y.; Aynaoglu, G.; Caglar, G. S.; Soylemez, F.Early second trimester retinol-binding protein-4 values in cases with or without gestational diabetes mellitus risk factors: a cross-sectional study [J].Journal of Obstetrics and Gynaecology Research, 2015, 41( (1)): 55-61
[5]Mallat, Z.; Simon, T.; Benessiano, J.; Clément, K.; Taleb, S.; Wareham, N. J.; Luben, R.; Khaw, K. T.; Tedgui, A.; Boekholdt, S. M.Retinol-binding protein 4 and prediction of incident coronary events in healthy men and women[J].Journal of Clinical Endocrinology & Metabolism, 2009, 94( (1)):255-260
[6]Tacke, F.; Weiskirchen, R.; Trautwein, C.Liver function critically determines serum retinol-binding protein 4 (RBP4) levels in patients with chronic liver disease and cirrhosis. [J].Hepatology, 2008, 48( (5)):1724-1725
[7]罗仍卓么; 王立贤; 孙世铎.北京黑猪RBP4基因与繁殖性状的关联分析[J].畜牧兽医学报, 2008 , 39 ((5)):536-539
[8] 刘璐; 梁永晔; 吴正常; 苏先敏; 郑先瑞; 王瑾; 吴圣龙; 包文斌.大白猪、长白猪和杜洛克猪RBP4基因多态性及其与繁殖性能的相关分析[J].中国畜牧兽医, 2012., 39( (10)):169-172
[9]Rajguru, S. U.; Kang, Y. H.; Ahluwalia, B. S.Localization of retinol (vitamin A) in rat testes[J].Journal of Nutrition, 1982, 112((10)):1881-1891
[10]Schmitt, M. C.; Ong, D. E. Expression of cellular retinol-binding protein and lecithin-retinol acyltransferase in developing rat testis. [J].Biology of Reproduction, 1993, 49 ( (5)):972-979
[11]Moura, A. A.; Jr, D. J.; Eberhardt, D. M.; Erickson, B. H.; Godkin, J. D.Immunolocalization of retinol-binding protein and profiles of its mRNA as related to testicular development in the beef bull.[J].Domestic Animal Endocrinology, 1998, 15( (1)):35-44
[12]Wemheuer, W.; Steinbrink, J.; Fuhrmann, H.; Schmidt, F. W.; Sallmannn, H. P.The effect of vitamin A and beta-carotene on the vitamin E status, ejaculation parameters and health of boar used for insemination].[J].Dtw Deutsche Tierrztliche Wochenschrift, 1996, 103 ((10) ):431-437
[13]Lin, C. L.; Ponsuksili, S.; Tholen, E.; Jennen, D. G. J.; Schellander, K.; Wimmers, K..Candidate gene markers for sperm quality and fertility of boar.[J].Animal Reproduction Science, 2006, 92( (3–4)):349-363
[14]Cheng, J.; Song, Z. Y.; Pu, L.; Yang, H.; Zheng, J. M.; Zhang, Z. Y.; Shi, X. E.; Yang, G. S.Retinol binding protein 4 affects the adipogenesis of porcine preadipocytes through insulin signaling pathways.[J].Biochemistry and cell biology = Biochimie et biologie cellulaire ;, 2013, 91( (4)):236-243
[15] 孙丽娜.视黄醇结合蛋白4的体外表达及其对猪卵母细胞成熟的影响[J].吉林大学, 2015, :1-40
[16]Livera, G.; Rouiller-Fabre, V.; Pairault, C.; Levacher, C.; Habert, R.Regulation and perturbation of testicular functions by vitamin A.[J].Reproduction, 2002, 124( (2)):173-180
[17]Livera, G.; Rouillerfabre, V.; Durand, P.; Habert, R.Multiple effects of retinoids on the development of Sertoli, germ, and Leydig cells of fetal and neonatal rat testis in culture.[J].Biology of Reproduction, 2009, 62 ((5)):1303-1314
[18]Livera, G.; Rouiller-Fabre, V.; Habert, R.Retinoid receptors involved in the effects of retinoic acid on rat testis development.[J].Biology of Reproduction, 2001, 64 ((5), ):1307-1314
[19]Mackenzie, S. H.; Roberts, M. P.; Liu, K. H.; Doré, J. J.; Godkin, J. D. Bovine endometrial retinol-binding protein secretion, messenger ribonucleic acid expression, and cellular localization during the estrous cycle and early pregnancy.[J].Biology of Reproduction ., 1998, 57 ((6)):1445-1450
[20]Abreu, R. D. S.; Penalva, L. O.; Marcotte, E. M.; Vogel, C.Global signatures of protein and mRNA expression levels.[J].Molecular Biosystems, 2009, 5 ((12)):1512-1526