Willis P, Sekhar K N, Brooks P, Fayrer-Hosken R A
Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602.
J Exp Zool. 1994 May 1;268(6):477-85. doi: 10.1002/jez.1402680608.
To characterize further the events involved in fertilization and early embryonic development in the mare, effect of the estrous cycle on oviductal fluid proteins was investigated. Five mares had indwelling cannulas placed in their oviducts so that fluid could be collected throughout the estrous cycle. Daily fluid volumes were recorded and mares were monitored for signs of standing estrus. Oviductal fluid samples were pooled across mares according to stage of cycle (either estrus or nonestrus) for further analysis. Two-dimensional polyacrylamide gel electrophoresis (PAGE) was used to determine proteins present in estrus-associated and nonestrus-associated equine oviductal fluid as compared to blood serum from the same mares. Oviductal fluid volumes increased significantly during estrus to an average of 3.94 ml/24 hr from 1.44 ml/24 hr during nonestrus. Total oviductal protein increased significantly from 24.6 mg/24 hr during nonestrus to 53.9 mg/24 hr during estrus. One-dimensional PAGE demonstrated that the proteins in equine oviductal fluid were present throughout the cycle and also common to equine serum. Reducing conditions revealed one band at 106 kDa detected only in nonestrus-associated oviductal fluid, while nonreducing conditions revealed bands at 48 and 25 kDa that were present in oviductal fluid in general. Two-dimensional PAGE demonstrated three 50 kDa proteins that were detected only in estrus-associated oviductal fluid and several 24 kDa proteins detected only in nonestrus-associated oviductal fluid. Those proteins found only in estrus-associated oviductal fluid may be vital to the fertilization process, while those found only in nonestrus-associated oviductal fluid may be vital to early embryonic development.
为了进一步描述母马受精和早期胚胎发育过程中的相关事件，研究了发情周期对输卵管液蛋白质的影响。五匹母马的输卵管内留置了插管，以便在整个发情周期收集液体。记录每日液体量，并监测母马的静立发情迹象。根据发情周期阶段（发情期或非发情期）将母马的输卵管液样本合并，用于进一步分析。使用二维聚丙烯酰胺凝胶电泳（PAGE）来确定发情期相关和非发情期相关的马输卵管液中存在的蛋白质，并与同一母马的血清进行比较。发情期输卵管液量显著增加，从非发情期的1.44毫升/24小时平均增加到3.94毫升/24小时。输卵管总蛋白从非发情期的24.6毫克/24小时显著增加到发情期的53.9毫克/24小时。一维PAGE显示，马输卵管液中的蛋白质在整个周期都存在，并且也是马血清中常见的。还原条件下在非发情期相关的输卵管液中仅检测到一条106 kDa的条带，而非还原条件下在一般输卵管液中检测到48 kDa和25 kDa的条带。二维PAGE显示，仅在发情期相关的输卵管液中检测到三种50 kDa的蛋白质，仅在非发情期相关的输卵管液中检测到几种24 kDa的蛋白质。那些仅在发情期相关的输卵管液中发现的蛋白质可能对受精过程至关重要，而那些仅在非发情期相关的输卵管液中发现的蛋白质可能对早期胚胎发育至关重要。
