Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (21): 4465-4474.doi: 10.3864/j.issn.0578-1752.2012.21.015

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Cloning, Expression and Bioinformatics Analysis of Porcine CatSperB and CatSperG Genes

 SONG  Cheng-Yi, ZHOU  Jia-Qing, FENG  Xiao-Jun, XIE  Yu-Xiu, LI  Qing-Ping, WU  Han, GAO  Bo, WANG  Xiao-Yan   

  1. 1.扬州大学动物科学与技术学院，江苏扬州225009
  • Received:2012-07-11 Online:2012-11-01 Published:2012-07-27

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Abstract: 【Objective】The aim of the current study is to confirm the existence of porcine CatSperB and CatSperG genes, and investigate the protein structures, evolutionary relationship and the spatial-temporal expression profiles of CatSperB and CatSperG. 【Method】The in silico and molecular cloning was used to identify the full length cDNAs of porcine CatSperB and CatSperG, and the spatial-temporal expression profile was investigated by qualitative and fluorescence quantitative RT-PCR. 【Result】The in silico transcripts of 3 508 bp CatSperB and 3 715 bp CatSperG were identified, and they contain 3 330 bp and 3 483 bp ORFs of CatSperB and CatSperG, respectively, and the sequences were confirmed by TA cloning. The sequence similarity of coding sequences (CDS) of porcine CatSperB and CatSperG with human, cattle, horse, and dog, and other animals was above 80%. CatSperB is a 125.79 kD and stable protein, while CatSperG is a 133.40 kD and unstable protein. Both CatSperB and CatSperG contain seven conservative trans-membrane domains, and a coiled-coil motif was also identified in the C terminal of CatSperG, but this motif was not found in CatSperB. The porcine CatSperB and CatSperG displayed higher degree of homology with the orthologs of cattle, dog and horse, and lower homology with those of human and mouse. The RT-PCR analysis showed that CatSperB and CatSperG were detected mainly in testis, but CatSperB also expressed in other tissues. The mRNA expression of CatSperB and CatSperG was significantly improved at the around stage of spermatogenesis (Day 60), puberty (Day 90) and sex maturity (Day 150) (P＜0.05). 【Conclusion】The cDNA clones of CatSperB and CatSperG, and a series of bioinformatics parameters of these proteins were obtained. Seven conservative trans-membrane domains in CatSperB and CatSperG and the homology among species were revealed. Furthermore, results of this study had proved that CatSperB and CatSperG expressed mainly in tests, and the change of mRNA expression was paralleled with sexual development of boar.

Key words: pig, CatSperB gene, CatSperG gene, cloning, spatial-temporal expression

[1]Liu J, Xia J, Cho K H, Clapham D E, Ren D. CatSperbeta, a novel transmembrane protein in the CatSper channel complex. The Journal of Biological Chemistry, 2007, 282(26): 18945-18952.

[2]Wang H, Liu J, Cho K H, Ren D. A novel, single, transmembrane protein CATSPERG is associated with CATSPER1 channel protein. Biology of Reproduction, 2009, 81(3): 539-544.

[3]Darszon A, Acevedo J J, Galindo B E, Hernández-González E O, Nishigaki T, Treviño C L, Wood C, Beltrán C. Sperm channel diversity and functional multiplicity. Reproduction, 2006, 131(6): 977-988.

[4]Darszon A, Labarca P, Nishigaki T, Espinosa F. Ion channels in sperm physiology. Physiological Reviews, 1999, 79(2): 481-510.

[5]Inaba K. Molecular architecture of the sperm flagella: molecules for motility and signaling. Zoological Science, 2003, 20(9): 1043-1056.

[6]Jimenez-Gonzalez C, Michelangeli F, Harper C V, Barratt C L, Publicover S J. Calcium signalling in human spermatozoa: a specialized ‘toolkit’ of channels,transporters and stores. Human Reproduction Update, 2006, 12(3): 253-267.

[7]张瑞芳, 曾海涛. 钙离子通道与精子运动功能. 生命的化学, 2009, 29(6): 878-882

Zhang R F, Zeng H T. Calcium channel and sperm motility. Chemistry of Life, 2009, 29(6): 878-882.

[8]Ren D, Navarro B, Perez G, Jackson A C, Hsu S, Shi Q, Tilly J L, Clapham D E. A sperm ion channel required for sperm motility and male fertility. Nature, 2001, 413(6856): 603-609.

[9]Qi H, Moran M M, Navarro B, Chong J A, Krapivinsky G, Krapivinsky L, Kirichok Y, Ramsey I S, Quill T A, Clapham D E. All four CatSper ion channel proteins are required for male fertility and sperm cell hyperactivated motility. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104(4): 1219-1223.

[10]孔丽娟, 杨媛媛, 王根林. CatSper与精子超活化. 中华男科学杂志, 2007, 13(2): 164-167.

Kong L J, Yang Y Y, Wang G L. CatSper and sperm hyperactivation. National Journal of Andrology, 2007, 13(2): 164-167.

[11]Xia J, Reigada D, Mitchell C H, Ren D. CATSPER channel-mediated Ca2+ entry into mouse sperm triggers a tail-to-head propagation. Biology of Reproduction, 2007, 77(3): 551-559.

[12]Carlson A E, Westenbroek R E, Quill T, Ren D, Clapham D E, Hille B, Garbers D L, Babcock D F. CatSper1 required for evoked Ca2+ entry and control of flagellar function in sperm. Proceedings of the National Academy of Sciences of the United States of America, 2003, 100(25): 14864-14868.

[13]Carlson A E, Quill T A, Westenbroek R E, Schuh S M, Hille B, Babcock D F. Identical phenotypes of CatSper1 and CatSper2 null sperm. The Journal of Biological Chemistry, 2005, 280(37): 32238-32244.

[14]Quill T A, Sugden S A, Rossi K L, Doolittle L K, Hammer R E, Garbers D L. Hyperactivated sperm motility driven by CatSper2 is required for fertilization. Proceedings of the National Academy of Sciences of the United States of America, 2003, 100(25): 14869-14874.

[15]Song C Y, Gao B, Wu H, Xie Y X, Wang X Y, Li B C, Chen G H, Mao J D. Molecular cloning, spatial and temporal expression analysis of CatSper genes in the Chinese Meishan pigs. Reproductive Biology and Endocrinology, 2011, 9: 132-143.

[16]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods, 2001, 25(4): 402-408.

[17]Poiani A. Complexity of seminal fluid: a review. Behavioral Ecology and Sociobiology, 2006, 60(3): 289-310.

[18]Töpfer-Petersen E, Romero A, Varela P F, Ekhlasi-Hundrieser M, Dostàlovà Z, Sanz L, Calvete J J. Spermadhesins: a new protein family. Facts, hypotheses and perspectives. Andrologia, 1998, 30(4/5): 217-224.

[19]Shivaji S, Scheit K H, Bhargava P M. Proteins of Seminal Plasma. Malden, MA: Wiley-Interscience Publication, 1990.

[20]Quill T A, Wang D, Garbers D L. Insights into sperm cell motility signaling through sNHE and the CatSpers. Molecular and Cellular Endocrinology, 2006, 250(1/2): 84-92.

[21]Lobley A, Pierron V, Reynolds L, Allen L, Michalovich D. Identification of human and mouse CatSper3 and CatSper4 genes: characterization of a common interaction domain and evidence for expression in testis. Reproductive Biology and Endocrinology, 2003, 1: 53-67.

[22]McFarlane A A, Orriss G L, Stetefeld J. The use of coiled-coil proteins in drug delivery systems. European Journal of Pharmacology, 2009, 625(1/3): 101-107.

[23]Li H G, Liao A H, Ding X F, Zhou H, Xiong C L. The expression and significance of CATSPER1 in human testis and ejaculated spermatozoa. Asian Journal of Andrology, 2006, 8(3): 301-306.

[24]Li H G, Ding X F, Liao A H, Kong X B, Xiong C L. Expression of CatSper family transcripts in the mouse testis during post-natal development and human ejaculated spermatozoa: relationship to sperm motility. Molecular Human Reproduction, 2007, 13(5): 299-306.

[25]Jin J L, O’Doherty A M, Wang S, Zheng H, Sanders K M, Yan W. CatSper3 and CatSper4 encode two cation channel-like proteins exclusively expressed in the testis. Biology of Reproduction, 2005, 73(6): 1235-1242.

[26]余  州, 张晓燕, 刁美玲, 王  勇, 唐爱发, 桂耀庭. CatSper基因家族在人和小鼠组织中的表达特征. 中国男科学杂志, 2009, 23(10): 1-4.

Yu Z, Zhang X Y, Diao M L, Wang Y, Tang A F, Gui Y T. The expression characteristics of CatSper gene family in human and mouse tissues. Chinese Journal of Andrology, 2009, 23(10): 1-4.

[27]Lunstra D D, Ford J J, Klindt J, Wise T H. Physiology of the Meishan boar. Journal of Reproduction and Fertility Supplement, 1997, 52: 181-193.
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