Effects of pH of Washing Solution, Washing Frequency and Individual Bull on Proton Concentration in the Sperm Washed Solution and Sperm Acrosome Reaction

精子 洗滌液의 pH와 洗滌頻度 및 수소個體가 精子洗滌液內 水素이온濃度와 소 精子의 尖帽反應에 미치는 影響

This study was carried out to investigate the effects of pH of sperm-washing solution, sperm-washing frequency and individual bull on concentration of hydrogen ion in sperm-washed solution and sperm acrosome reaction. The results obtained were as follow. 1. When bovine sperm was washed at 4 times with SHP solution and incubated, the difference of light absorbance between sperm-washing and sperm-washed solution(${\Delta}$Ao-${\Delta}$At) was higher in 2nd sperm-washed solution than that in the other washed solutions. 2. When sperm was thrice washed with SHP solutions of pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45, and 8.83, ${\Delta}$Ao-${\Delta}$At was significantly increased at pH7.69 to 8.83, and ${\Delta}$Ao-${\Delta}$At in 1st sperm washed solution was significantly higher than that in 2nd and 3rd sperm washed solution. 3. When sperm of Holstein, KNC and Hereford was thrice washed with SHP solutions of pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45, and 8.83, Holstein showed higher ${\Delta}$Ao-${\Delta}$At of sperm Washed solution than KNC and Hereford, and ${\Delta}$Ao-${\Delta}$At in sperm washed solution was significantly increased at 7.69 for Holstein and at 8.15 for KNC and Hereford, respectively. 4. When sperm was thrice with SHP solution of pH 6.8, 7.1 and 7.4, and then incubated in mTALP of pH 7.4 for 15 minutes, in 1st and 2nd sperm washing ${\Delta}$Ao-${\Delta}$At of sperm washed solution was significantly higher at pH 7.1 and 7.4 than at pH 6.8, and the sperm acrosome reaction of pH 6.8, 7.1 and 7.4 was 49.1, 68.8 and 72.9%, respectively. The sperm acrosome reaction of pH 7.1 and 7.4 was higher than that of pH 6.8.

본 實驗은 精子 洗滌液의 pH와 洗滌頻度 및 수소개체가 精子洗滌液內 水素이온과 精子尖帽反應에 미치는 影響을 究明하기 위하여 실시하였는바 그 결과는 다음과 같다. 1. 精子를 SHP液으로 4차례 反復洗滌하여 培養하였을 때, 洗滌液의 吸光度差間 變化는 2차 精子洗滌液에서 가장 높았다. 2. 精子를 pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45 및 8.83 SHP液으로 3번 洗滌하였을 때, 精子洗滌液의 吸光度差間 變化는 pH 7.69~8.83에서 유의하게 증가하였으며, 1차 精子洗滌液이 2차와 3차 精子洗滌液보다 유의하게 높았다. 3. Holstein, KNC 및 Hereford 精子를 pH 5.99, 6.38, 6.78, 7.10, 7.40, 7.69, 8.15, 8.45 및 8.83의 SHP液으로 3번 洗滌하였을 때, 精子洗滌液의 光度差間 變化는 Holstein이 KNC와 Hereford보다 유의하게 높았으며, Holstein은 pH 7.69~8.83에서, KNC과 Hereford는 8.15에서 각각 유의하게 증가하였다. 4. 精子를 pH가 6.8, 7.1 및 7.4인 SHP液으로 3번 洗滌한 다음 pH 7.4인 mTALP液에서 15분간 培養하였을 때, 精子洗滌液의 吸光度差間 變化는 1차 및 2차 精子洗滌에서 pH 7.1과 7.4가 6.8보다 유의하게 높았으며, 精子尖帽反應率은 pH 7.1 및 7.4가 각각 68.8 및 72.9%로 pH 6.8의 49.1%보다 유의하게 높았다(P<0.05).