원문정보
초록
영어
Sperm decondensation must be accomplished before fluorescence in situ hybridization (FISH). However, the lack of studies were conducted to discover the optimal porcine sperm decondensation for improving efficiency of FISH. We therefore sought to investigate the proper conditions of porcine sperm decondensation for FISH. In this trial, semen samples were obtained from eleven boar and immediately washed. After washing, the suspension was smeared and fixed on slides. Air-dried slides were incubated at 37℃ for 0, 15, 30 and 45 min in a 5 mmol/L dithiothreitol (DTT) to cleave protamine disulfide bonds, and thus sperm head decondensation was occurred. Subsequently, slides were counter-stained with hoechst 33258 to measure several parameters of sperm head by using microscope. The area, perimeter, and length of sperm head were significantly increased after 30 min incubation. This result demonstrated that sperm head is effectively expanded, which is essential for sperm decondensation after 30 min incubation. Subsequently, the roundness of sperm head was significantly increased after 15 min incubation. This result showed that the circularity of sperm head, which is important for sperm head decondensation, is ideal after 15 min. Taken together, the incubation time of optimal sperm head decondensation could be 30 min to get better achievement for FISH.