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http://dx.doi.org/10.12750/JET.2017.32.3.227

Comparative Evaluation on Sperm Parameter of Transgenic Pigs with General Pigs  

Park, Sang Hyoun (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Lee, Gunsup (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Lee, Joo Yung (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Kim, Kyung Woon (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Byun, Sung-June (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Ock, Sun A (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Hwang, Seongsoo (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Yang, Hyeon (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Woo, Jae-Seok (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Oh, Keon Bong (Animal Biotechnology Division, National Institute of Animal Science, RDA)
Publication Information
Journal of Embryo Transfer / v.32, no.3, 2017 , pp. 227-233 More about this Journal
Abstract
Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.
Keywords
Acrosome integrity; Morphology; Motility; Transgenic pigs; Viability;
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Times Cited By KSCI : 5  (Citation Analysis)
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1 Zou CX and Yang ZM. 2000. Evaluation on sperm quality of freshly ejaculated boar semen during in vitro storage under different temperatures. Theriogenology 53:1477-1488.   DOI
2 Park SH, and Yu IJ. 2015. Evaluation of toxicity of green tea extract in chilled boar spermatozoa. J. Emb. Trans. 30:1-6.   DOI
3 Pursel VG, Bolt DJ, Miller KF, Pinkert CA, Hammer RE, Palmiter RD and Brinster RL. 1990. Expression and performance in transgenic pigs. J. Reprod. Fertil. Suppl. 40:235-245.
4 Rahman Md A, Park SH, and Yu IJ. 2016 Effect of monosaccharides in glycerol-free tris extender on reactive oxygen species and apoptosis in dog sperm cryopreservation. Cryoletters. 38:51-57.
5 Rexroad CE Jr, Hammer RE, Bolt DJ, Mayo KE, Frohman LA, Palmiter RD and Brinster RL. 1989. Production of transgenic sheep with growth-regulating genes. Mol. Reprod. Dev. 1:164-169.   DOI
6 Waberski D, Magnus F, Mendonca Ferreira F, Petrunkina AM, Weitze KF and Topfer-Petersen E. 2005. Importance of sperm-binding assays for fertility prognosis of porcine spermatozoa. Theriogenology 63:470-484.   DOI
7 Wall RJ, Pursel VG, Shamay A, McKnight RA, Pittius CW and Hennighausen L. 1991. High-level synthesis of a heterologous milk protein in the mammary glands of transgenic swine. Proc. Natl. Acad. Sci. U S A. 88:1696-1700.   DOI
8 Wall RJ, Pursel VG, Hammer RE and Brinster RL. 1985. Development of porcine ova that were centrifuged to permit visualization of pronuclei and nuclei. Biol. Reprod. 32:645-651.   DOI
9 Weitze KF. 2012. The importance of boar sperm motility and morphology for fertility, International Pig Topics. East Yorkshire 27:13-15.
10 Wolf J. 2009. Genetic parameters for semen traits in AI boars estimated from data on individual ejaculates. Reprod. Domest. Anim. 44:338-344.   DOI
11 Zheng ZY, Oh KB, Koo DB, Han YM and Lee KK. 2008. Expression of the Transgene is Consistently Inherited to High Numbers of Generations and Independent on Its Source. Reprod. Dev. Biol. 32:39-43
12 Hwang IS, Lee SC, Kim SW, Kwon DJ, Park MR, Yang H, Oh KB, Ock SA, Woo JS, Im GS and Hwang SS. 2017. Analysis of Semen Parameters in ${\alpha}1$,3-Galactosyltransferase-/- Boars. J. Emb. Trans. 32:53-58.   DOI
13 Hwang S, Oh KB, Kwon D-J, Ock S-A, Lee J-W, Im G-S, Lee S-S, Lee K and Park J-K. 2013. Improvement of cloning efficiency in minipigs using post-thawed donor cells treated with roscovitine. Molecular biotechnology 55:212-216.   DOI
14 Johnson LA, Weitze KF, Fiser P and Maxwell WM. 2000. Storage of boar semen. Anim. Reprod. Sci. 62:143-172.   DOI
15 Kato Y, Tani T, Sotomaru Y, Kurokawa K, Kato J, Doguchi H, Yasue H and Tsunoda Y. 1998. Eight calves cloned from somatic cells of a single adult. Science 282:2095-2098.   DOI
16 Ko N, Lee J-W, Hwang SS, Kim B, Ock SA, Lee S-S, Im G-S, Kang M-J, Park J-K and Jong Oh S. 2013. Nucleofection-Mediated ${\alpha}1$, 3-galactosyltransferase Gene Inactivation and Membrane Cofactor Protein Expression for Pig-to-Primate Xenotransplantation. Animal biotechnology 24:253-267.   DOI
17 Hammer RE, Pursel VG, Rexroad CE Jr, Wall RJ, Bolt DJ, Ebert KM, Palmiter RD and Brinster RL. 1985. Production of transgenic rabbits, sheep and pigs by microinjection. Nature 315:680-683.   DOI
18 Lee SC, Lee HS, Oh KB, Hwang IS, Yang H, Park MR, Ock SA, Woo JS, Im GS and Hwang SS. 2017. Production and Breeding of Transgenic Cloned Pigs Expressing Human CD73. Dev. Reprod. 21:157-165.   DOI
19 Maleszewski M, Kuretake S, Evenson D, Yanagimachi H, Bjordahl J and Yanagimachi R. 1998. Behavior of transgenic mouse spermatozoa with galline protamine. Biol. Reprod. 58:8-14.   DOI
20 Meliska CJ and Bartke A. 1997. Copulatory behavior and fertility in transgenic male mice expressing human placental growth hormone gene. J. Androl. 18:305-311.
21 Britt JH, Almond GW and Flowers WL. 1999. Diseases of the reproductive system. In: Strae B, D'Allaire S, Mengeling W, Taylor D (eds), Disease of Swine, 8th edn. Blackwell Science Ltd, Ames, IA, p. 905.
22 Ahn KS, Kim YJ, Kim M, Lee BH, Heo SY, Kang MJ, Kang YK, Lee JW, Lee KK, Kim JH, Nho WG, Hwang SS, Woo JS, Park JK, Park SB and Shim H. 2011. Resurrection of an ${\alpha}$-1,3-galactosyltransferase gene-targeted miniature pig by recloning using postmortem ear skin fibroblasts. Theriogenology 75:933-939.   DOI
23 Alm K, Peltoniemi OA, Koskinen E and Andersson M. 2006. Porcine field fertility with two different insemination doses and the effect of sperm morphology. Reprod. Domest. Anim. 41:210-213.   DOI
24 Bartke A, Naar EM, Johnson L, May MR, Cecim M, Yun JS and Wagner TE. 1992. Effects of expression of human or bovine growth hormone genes on sperm production and male reproductive performance in four lines of transgenic mice. J. Reprod. Fertil. 95:109-118.   DOI
25 Cooper DK. 2012. A brief history of cross-species organ transplantation. Proc (Bayl Univ Med Cent) 25:49-57.   DOI
26 Costantini F and Lacy E. 1981. Introduction of a rabbit beta-globin gene into the mouse germ line. Nature 294:92-94.   DOI
27 Flowers WL. 1997. Management of boars for efficient semen production. J. Reprod. Fertil. Suppl. 52:67-78.
28 Gadea J, Matas C and Lucas X. 1998. Prediction of porcine semen fertility by homologous in vitro penetration (hIVP) assay. Anim. Reprod. Sci. 54:95-108.   DOI
29 Gordon JW and Ruddle FH. 1981. Integration and stable germ line transmission of genes injected into mouse pronuclei. Science 214:1244-1246.   DOI
30 Gadea J, Selles E, and Marco MA. 2004. The predictive value of porcine seminal parameters on fertility outcome under commercial conditions. Reprod. Domest. Anim. 39:303-308.   DOI
31 황성수, 오건봉, 김동훈, 우제석, 심호섭, 윤익진, 박진기, 임기순. 2012. ${\alpha}1$, 3-Galactosyltransferase (GalT) 유전자가 완전 Knock-out (-/-) 된 바이오장기용 형질 전환 돼지 생산. 한국수정란이식학회지. 27:9-14.
32 배성훈, 양병철, 고응규, 오건봉, 성환후, 민관식, 박응우, 박수봉, 황성수. 2009. 체세포 복제 한우 수송아지의 성장 특성과 번 식생리적 변화. 한국수정란이식학회지 24:177-182.
33 배성훈, 황성수, 양병철, 고응규, 김동훈, 임기순, 최화식, 진동일, 양보석, 성환후. 2007. 체세포 복제 한우 수소의 정액 성상, 정자의 활동성 및 수정 능력 분석. 한국동물번식학회지 31:139-143.
34 이연근, 박진기, 민관식, 이창현, 성환후, 전익수, 임석기, 양병철, 임기순, 장원경, 김진회, 이훈택, 정길생. 2002. 사람 조혈인자 유전자 (Human Erythropoietin Gene)를 도입한 형질전환돼지 생산. 한국동물번식학회지. 26:95-104.
35 Aboagla EM and Terada T. 2003. Trehalose-enhanced fluidity of the goat sperm membrane and its protection during freezing. Biol. Reprod. 69:1245-1250.   DOI