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Analysis of Frozen-Thawed Sperm Characteristic in Alpha 1,3-Galactosyltransferase(GalT) Knock-out Cloned Miniature Pig  

Woo, Jea-Seok (National Institute of Animal Science, RDA)
Lee, Yong-Seung (College of Animal Life Sciences, Kangwon National University)
Yoo, Han-Jun (College of Animal Life Sciences, Kangwon National University)
Hwang, Seong-Soo (National Institute of Animal Science, RDA)
Oh, Keon-Bong (National Institute of Animal Science, RDA)
Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University)
Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University)
Park, Soo-Bong (National Institute of Animal Science, RDA)
Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.35, no.3, 2011 , pp. 385-390 More about this Journal
Abstract
The purpose of this study was undertaken to evaluate of cryopreservation efficiency in ${\alpha}$ 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. After thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.
Keywords
${\alpha}$ 1,3-Galactosyltransferase knock out(GalT KO); Cloned miniature pig; Cryopreservation; Sperm ability; Duroc; PWG miniature pig;
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Times Cited By KSCI : 3  (Citation Analysis)
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1 Potter WL, Upton PC, Dunn BL (1979): Morphological changes as observed by light microscopy of the acrosome of boar spermatozoa subjected to deep freezing. Aust J Biol Sci 32:575-578.
2 Pursel VG, Johnson LA, Rampacek GB (1972) Acrosome morphology of boar spermatozoa incubated before cold shock. J Anim Sci 34:278-283.
3 Wang WH, Abeydeera LR, Fraser LR, Niwa K (1995): Functional analysis using chlortetracycline fluorescence and in vitro fertilization of frozen thawed ejaculated boar spermatozoa incubated in proteinfree chemically defined medium. J Reprod Fertil 104: 305-313.   DOI
4 김성곤, 장현용, 박동헌, 박춘근, 정희태, 김정익, 양부근 (2006): 돼지 정액의 간편 동결 방법 확립과 동결정액의 융해 후 생존성 평가. 한국동물번식학회지30:59-64.
5 이용승, 최원철, 이승형, 정희태, 이상영, 양부근, 박춘근 (2006): Miniature Pig와 Duroc 종간의 동결-융해후 정액 성상 비교. 한국수정란이식학회지 21:263-271.
6 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, 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   ScienceOn
7 Cooper DKC, David KC (2006): Xenotransplantation: the road ahead. Curr Opin Organ Transplant 11: 151-153.   DOI   ScienceOn
8 Cooper DKC, Koren E, Oriol R (1993): Genetically engineered pigs. Lancet 342:682-683.   DOI   ScienceOn
9 Darin-Bennett A, White IG (1977): Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold shock. Cryobiology 14:466- 470.   DOI   ScienceOn
10 Eriksson BM, Petersson, Rodroguez-martinez H (2002): Field fertility with exported boar semen frozen in the new Flac Pack container. Theriogenology 58: 1065-1079.   DOI   ScienceOn
11 Lee SH, Cheong HT, Yang BK, Park CK (2005): Development of semen extenders by assessment of sperm viability in miniature pig semen. Reprod Dev Bio 29:247-252.
12 Gillan L, Evans G, Maxwell WMC (1997): Capacitation status and fertility of fresh and frozen thawed ram spermatozoa. Reprod Fertil Dev 9:481- 487.   DOI   ScienceOn
13 Good AH, Cooper DKC, Malcolm AJ, Ippolito RM, Koren E, Neethling FA, Ye Y, Zuhdi N, Lamontagne LR (1992): Identification of carbohydrate structures that bind human antiporcine antibodies: implications for discordant xenografting in humans. Transplant Proc 24:559-562.
14 Holt WV (2000): Basic aspects of frozen storage of semen. Anim Reprod Sci 62:3-22.   DOI   ScienceOn
15 Lin DS, Connor WE, Wolf DP, Neuringer M, Hachey DL (1993) Unique lipids of primate spermatozoa- desmosterol and docosahexaenoic acid. J Lipid Res 34:491-499.
16 Loomis PB, Graham JK (2008): Commercial semen freezing: individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols. Anim Reprod Sci 105:119-128.   DOI
17 Maxwell WMC, Johnson LA (1997): Membrane status of boar spermatozoa after cooling or cryopreservation. Theriogenology 48:209-219.   DOI   ScienceOn
18 Mazur P (1984): Freezing of living cells: mechanisms and implications. Am J Physiol 247:C125-C- 142.
19 Neild DM, Gadella BM, Maria GC, Marcelo H, Miragaya C, Colenbrander B, Aguero A (2003): Membrane changes during different stages of a freeze thaw protoxol for equine semen cryopreservation. Theriogenology 59:1693-1705.   DOI   ScienceOn
20 Park JE, Lynch DV (1992): Lipid-composition and thermotropic phase-behavior of boar, bull, stallion and rooster spermatozoa. Cryobiology 29:255-266.   DOI   ScienceOn
21 Phelps CJ, Koike C, Vaught TD, Boone J, Wells KD, Chen SH, Ball S, Specht SM, Polejaeva IA, Monahan JA, Jobst PM, Sharma SB, Lamborn AE, Garst AS, Moore M, Demetris AJ, Rudert WA, Bottino R, Bertera S, Trucco M, Starzl TE, Dai Y, Ayares DL (2003): Production of alpha 1,3-galactosyltransferase-deficient pigs. Science 299:411-414.   DOI   ScienceOn
22 Polge C, Salamon S, Wilmut I (1970): Fertilizing capacity of frozen boar semen following surgical insemination. Vet Res 87:424-428.