Browse > Article

Effects of Methyl-Beta-Cyclodextrin (MBCD) on Cold Shock and Membrane Cholesterol Quantity during the Freezing Process of Miniature Pig Spermatozoa  

Lee, Sung-Young (College of Animal Life Sciences, Kangwon National University)
Lee, Yong-Seung (College of Animal Life Sciences, Kangwon National University)
Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University)
Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University)
Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
Publication Information
Reproductive and Developmental Biology / v.35, no.3, 2011 , pp. 265-271 More about this Journal
Abstract
This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until $5^{\circ}C$ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.
Keywords
Miniature pig; Cryopreservation; Cyclodextrin; Cholesterol;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Almlid T, Hofmo PO (1996): A brief review of frozen semen application under Norwegian AI service conditions. Reprod Domest Anim 31:169-173.
2 Amirat L, Tainturier D, Jeanneau L, Thorin C, Gerard O, Courtens JL, Anton M (2004): Bull semen in vitro fertility after cryopreservation using egg yolk LDL: A comparison with optidyl, a commercial egg yolk extender. Theriogenology 61:895-907.   DOI   ScienceOn
3 Crowe JH, Hoekstra FA, Crowe LM, Anchordoguy TJ, Drobnis E (1989): Lipid phase transitions measured in intact cell with Fourier transform infrared spectroscopy. Cryobiology 26:76-84.   DOI   ScienceOn
4 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
5 Hartel S, Diehl HA, Ojeda SF(1998): Methyl-beta-cyclodextrins and liposomes as water-soluble carriers for cholesterol incorporation into membranes and its evaluation by a microenzymatic fluorescence assay and membrane fluidity-sensitive dyes. Anal Biochem 258:277-284.   DOI   ScienceOn
6 Holt WV, North RD (1984): Partially irreversible cold-induced lipid phase transitions in mammalian sperm plasma membrane domains : freeze-fracture study. J Exp Zool 230:473-483.   DOI   ScienceOn
7 Holt WV, Head MF, North RD (1986): Thermotropic phase transitions in the plasma membrane of ram spermatozoa. J Reprod Fertil 78:445-457
8 Holt WV (1997): Alternative strategies for the longterm preservation of spermatozoa. Reprod Fertil Dev 9:309-319.   DOI   ScienceOn
9 Holt WV (2000): Basic aspects of frozen storage of semen. Anim Reprod Sci 62:3-22.   DOI   ScienceOn
10 Jeong YJ, Kim MK, Song HJ, Kang EJ, Ock SA, Mohana Kumar B, Balasubramanian S, Rho GJ (2009): Effect of $\alpha$-tocopherol supplementation during boar semen cryopreservation on sperm characteristics and expression of apoptosis related genes. Cryobiology 58:181-189.   DOI   ScienceOn
11 Purdy PH, Graham JK (2004): Effect of adding cholesterol to bull sperm membranes on sperm capacitation, the acrosome reaction, and fertility. Biol Reprod 71:522-527.   DOI   ScienceOn
12 Watson PF (1981): The effects of cold shock on sperm cell membranes. In: Morris GJ, Clarke A (eds.), Effects of Low Temperatures on Biological Membranes. London: Academic Press; 189-218.
13 White IG (1993): Lipids and calcium uptake of sperm in relation to cold shock and preservation: a review. Reprod Fertil Dev 5:639-658.   DOI   ScienceOn
14 Yeagle PL (1985): Cholesterol and the cell membrane. Biochim Biophys Acta 822:267-287.   DOI   ScienceOn
15 Pike LJ, Miller JM (1998): Cholesterol depletion delocalizes phosphatidyl inositol bisphosphate and inhibits hormone-stimulated phosphatidyl inositol turnover. J Biol Chem. 273:22298-22304.   DOI
16 Polge C, Salamon S, Wilmut I (1970): Fertilizing capacity of frozen boar semen following surgical in semination. Vet Res 87:424-428.
17 Lee SH, Cheong HT, Yang BK, Park CK (2005): Development of semen extenders by asessment of sperm viability in miniature pig semen. Reprod Dev Biol 29(4): 247-252.
18 Pursel VG, Johnson LA (1973): Procedure for the preservation of boar spermatozoa by freezing, USDA. ARS Bull 44-227:1-5.
19 Salisbury GW, Van Demark NL, Lodge JR (1978): Physiology of Reproduction and Artificial Insemination of Cattle. W. H. Freeman and Co. 2nd (eds), San Francisco, USA.
20 Johnson LA, Weitze KF, Fiser P, Maxwell WMC (2000): Storage of boar semen. Anim Reprod Sci 62:143- 172.   DOI   ScienceOn
21 Maxwell WMC, Johnson LA (1997): Membrane status of boar spermatozoa after cooling or cryopreservation. Therogenology 48:209-219.   DOI   ScienceOn
22 Mercado E, De, Hernandez M, Sanz E, Rodriguez A, Gomez E, Vazquez JM, Martinez EA, Roca J (2008): Evaluation of L-glutamine for cryopreservation of boar spermatozoa. Anim Reprod Sci 115:149-157.
23 Park JE, Lynch DV (1992): Lipid composition and thermortopic phase behavior of boar, bull, stallion and rooster sperm membranes. Cryobiology 29:255- 266.   DOI   ScienceOn
24 McGrath JJ (1988): Membrane transport properties. In: McGrath JJ, Diller KR, eds. Low Temperature Biotechnology: Emerging Applications and Engineering Contribution. New York, NY: ASME Press, BED-Vol 10, HTD-Vol 98, 273-330.