Effects of Individual Variance of Bull, Sperm Type and Pretreatment of Sperm and Oocytes on Male Pronuclear Formation and Developmental Rates in Korean Natitive Cattles

한우에 있어서 숫소 개체, 정자의 형태, 정자와 난자의 전처리 등이 ICSI후 웅성전핵 형성과 체외발생에 미치는 영향에 관한 연구

This study was carried out to investigate the effects of the bull, sperm type and sperm pretreatment on the pronuclear formation and in vitro development after injection of spermatozoa into in vitro matured bovine oocytes. 1. Spermatozoa derived room four bulls(A, B, C and D) were used for ICSI. The male pronuclear formation and developmental rates were 73.9∼87.0% and 33.3∼60.9%, respectively. 2. The effects of sperm type were examined. Male pronuclear formation rates by using fresh-and frozen-sperm, tail-cutting and tail-scoring sperm were 82.0%. 78.0%. 42.2% and 51.1% (p<0.05) while development rates were 56.0%. 42.0%, 17.8% and 22.2%, respectively. Fresh sperm achived a high mail pronuclear- and development rates than those of other groups. 3. Cheroical pretreatments were tested and compared. When sperm were pretreated with heparin, BFF(bovine follicula fluid), His, Ca Ionophore(I) and I + caffeine, mate pronuclear formation and developmental rates were 66.7∼82.2% and 33.3∼60.6%. respectively. and these values of treatment of I + caffeine were higher than that of other methods. 4. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated with or without zona pellucida were 80.0%. 72.0% and 46.0%, 36.0%, respectively.

본 연구는 숫소 개체별, 정자의 형태, 정자의 전처리 및 난자의 투명대부착과 미부착별로 현미조작에 의해 난자의 세포질내 단일정자를 주입했을 때 웅성전핵 형성율과 체외발생율을 조사하였다. 1. 숫소 개체별 정자를 이용하여 ICSI를 하였을 때 웅성전핵 형성율은 각각 73.9∼87.0%였고 체외발생율은 33.3∼60.9%를 나타냈다. 2. 신선 및 동결정자와 미부절단 정자와 두부, 미부손상 정자를 이용하여 ICSI를 하였을 때 웅성전핵 형성율은 각각 82.0%, 78.0% 42.2%, 51.1%였고, 체외발생율은 56.0%, 42.0%, 17.8%, 22.2%로서 신선정자를 이용했을 때가 다른 처리군에 비해 높은 전핵형성율과 체외발생율을 나타냈다. 3. 정자를 heparin, BFF, His, Ca Ionophore, I + caffeine 액으로 처리후 ICSI를 하였을 때 전핵 형성율은 66.7∼82.2%였고, 체외발생율은 33.3∼60.6%로서 I + caffeine 처리법이 다른 처리군에 비해 높게 나타났다. 4. 투명대 부착 난자 및 미부착 난자로 ICSI를 하였을 때 전핵형성율은 각각 80.0%, 72.0%였고, 체외발생율은 46.0%, 36.0%로서, 토명대부착 난자가 투명대 미부착 난자에 비해 높은 웅성전핵 형성율과 체외발생율을 나타냈다.