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題 名 | 鈣離子通道-Ryanodine接受體之遺傳型對豬卵母細胞體外成熟、受精與致活之影響=Effects of Genotype of Calcium Release Channel-Ryanodine Receptor on in Vitro Maturation, Fertilization and Activation of Porcine Oocytes |
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作 者 | 陳銘正; 林佳靜; 林育安; 李盈漢; 吳伊倫; 林明怡; 蔡宛蓁; | 書刊名 | 宜蘭大學生物資源學刊 |
卷 期 | 4:2 2008.12[民97.12] |
頁 次 | 頁135-140 |
分類號 | 437.653 |
關鍵詞 | 鈣離子通道; 豬卵母細胞; 體外成熟; 體外受精; 致活作用; Calcium release channel; Porcine oocyte; In vitro maturation; In vitro fertilization; Activation; |
語 文 | 中文(Chinese) |
英文摘要 | The present study examined the effects of genotype of calcium release channel -Ryanodine receptor (RyR) on in vitro maturation (IVM), fertilization (IVF) and activation of porcine oocytes. All oocyte-cumulus complexes were collected from 69 gilts' ovaries and cultured in M199 mediurn supplemented with gonadotrophin at 39℃ in 5 % CO2 in humidified air for 44-48 hours. After maturation culture, the oocytes were penetrated with sperm at 10^6/ml concentration for 6-8 hours of incubation or activated at field strength of 1.2 kv/cm, and then the oocytes were transferred into NCSU-23 medium for another 10 or 16 hours of incubation. Genomic DNA was isolated from gilts' ovaries and genotypes of RyR were analyzed by polymerase chain reaction and restriction fragment length polymorphism. Analysis of RyR genotypes showed that 85.5 and 14.5 % of gilts exhibited normal homozygous (C/C) and heterozygous (C/T) genotypes, respectively. No T/T genotype of RyR was identified. The frequency of T allele was 7.25 %. The results of IVF, IVF and activation showed that the maturation, fertilization and activation rate of oocytes from all ovaries were 68.2±19.6, 74.3±23.1 and 86.0±14.3%, respectively. There were no differences between C/C (70.0±15.8, 71.2±23.1 and 85.8±14.5%) and C/T (66.7±24.2, 85.1±22.3 and 86.5±10.4%) gilts (P>0.05). However, the polysperrny rate and male pronucleus(ei) formation of oocytes from C/T gilts (77.4±31.5 and 52.0±20.0%) were significantly higher (P<0.05) than those from C/C gilts (56.7±19.7 and 22 .4±15.0%), respectively. In conclusion, there was few gilts with T/T genotype, and no difference in maturation, fertilization, and activation of oocytes between C/C and C/T genotypes. However, the polyspermy rate and male pronucleus(ei) formation of oocytes were influenced by these RyR genotypes. |
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