To make up the medium for quantitative selection of capacitated-sperm through sucrose layer, the effects of BSA, caffeine, heparin and progesterone on sperm swim-up migration and movement were examined. And the results obtained were as follows; 1. BSA of 4mg/ml in bMSS stimulated sperm migration and movement, and attracted capacitated-sperm. 2. Caffeine of 5mM in bMSS containing 4mg/ml BSA stimulated sperm movement and attracted capacitated-sperm. 3. Heparin of 20/ml in bMAA containing both 4mg/ml BSA and 5mM caffeine stimulated movement and capacitation of sperm. 4. Progesterone of 50/ml in bMSS containing all 4mg/ml BSA, 5mM caffeine and 20/ml heparin (BCHP-MSS) attracted capacitated-sperm. 5. Effect of BCHP-MSS on sperm on sperm attraction was not different from effect of 10% follicular fluid solution (FF-MSS) on sperm swim-up separation. In conclusion, bMSS with 4mg/ml BSA, 5mM caffeine, 20/ml heparin and 50/ml progesterone(BCHP-MSS) was a optimal condition for selection of capacitated-sperm through sucrose layer.
난포액에 함유되어 있는 steroids와 sterol이 수정에 참여하는 정자의 주화성에 미치는 영향을 밝히기 위하여, progesterone, estradiol 17 beta 및 cholesterol이 sucrose 층으로부터 정자의 swim-up 분리에 미치는 영향을 조사하였던 바 얻어진 결과는 다음과 같다. 1. Progesterone은 정자의 이동과 운동성을 억제하였으나 수정능획득한 정자를 유인하였으며, 특히 50g/ml 수준의 progestero
The aim of this study is to elucidate sperm chemotaxis and to set up the optirnal condition for selection of motile and capacitated sperm from hovine frozen-semen. Thus, the effects of semen-washing after thawing, concentrations of progesterone (P4) and bovine serum albumin (BSA), and sperm-washing frequency on sperm selection were examined. For evaluating their effects, number, viability and acrosome reaction of sperm swim-up seperated from semen, which were incubated for 30 minutes at 36 in the M2 solution containing P4 and BSA, were investigated. For frozen-semen just after thawing, sperm recovery and viability were not significantly different between P4-treated and -untreated semen. However, washing frozen-semen decreased the number of sperm and increased the viability of sperm that were recovered from semen treated with P4. Progesterone affected the recovery rate, the viability and the acrosome-reaction rate of sperm recovered from washed frozen-semen. Especially, number of motile and capacitated sperm were highest in semen treated with 50g /ml among 0, 20, 50 and 100g /ml of P4 concentrations. BSA affected the recovery rate and the viability of sperm recovered from washed frozen-semen that were treated with 50g /ml of P4. Especially, the percentage of viable sperm were highest in semen treated with 4mg /ml among 0, 2, 4, and 6mg /ml of BSA concentrations. Repeatedly sperm-washing did not affect the recovery rate and the viability of sperm recovered from washed frozen-semen that were treated with 50g /ml of P4 and 4mg /ml of BSA In conclusion, using progesterone and BSA could efficiently make the selection of motile and capacitated sperm from washed frozen-semen.
Plasma progesterone concentrations were measured by using radjoimmunoassay for early diagnosis of pregnancy in Cheju-native mares. A total of 226 pony mares were examined for pregnancy during breeding and non breeding seasons. Plasma progesterone levels 20~23 days after the onset of oestrus were 4.67+O.67ng /rnl and O.55+O.O4ng /ml for mares becornrning pregnant and not pregnant after the estrus, respectively, and there was a significant differences (p<0.01) between the two groups. Progesterone concentration of pregnant mares gradually increased in 30 days andreached a peak (10.3ng /ml) during the third month of gestation. However, the concentration decresed to the base line (1.llng /rnl) at 7 months and gradually increased again as foaling approached (2.lng /ml). Early diagnosis for pregnancy of Cheju mares by progesterone level at 20~23 days after onset of oestrus was 88% accurate when 4.6ng /ml was used to classify mares as pregnancy and below 1.3ng /rnl was used to determine nonpregnant mares. However, the accuracy of the diagnosis was improved to 96% when a progesterone level of above 2ng /mi was used to classify mares for pregnancy. Diagnosis for pregnancy was 69.6% accurate when mares were classified as pregnancy by horse owners during breeding season. The progesterone levels of pregnant and non-pregnant mares during non-breeding season varied greatly between individual animals, Plasma progesterone levels of pregnant animals ranged from 3.5ng /mi to above 6.2ng /mi whereas similar values were observed in non-pregnant animals. Radioirnrnunoassay technicjues can be applied for the early pregnant diagnosis of Cheju native mares when progesterone levels are measured during the early gestation period (18~23 days after onset of oestrus). However, progesterone concentration of mares in non-breeding season is conisidered unsuitable as a indicator of pregnant diagnosis.