Kawakami E, Arai T, Oishi I, Hori T, Tsutsui T
Department of Reproduction, Nippon Veterinary and Animal Science University, Tokyo, Japan.
J Vet Med Sci. 2000 Jan;62(1):65-8. doi: 10.1292/jvms.62.65.
Ejaculated sperm collected from 12 beagle dogs were incubated in canine capacitation medium (CCM), supplemented with 5 microg/ml chondroitin sulfate A (CS), 5 microg/ml hyaluronic acid (HA), or 5 microg/ml heparin (HP) for 7 hr at 38 degrees C in a 5% CO2 in air atmosphere to investigate the effects of glycosaminoglycans (GAGs) on dog sperm capacitation. The percentages of motile sperm, hyperactivated sperm (%HY), and acrosome-reacted sperm (%AR) in all media were examined after 4 hr and 7 hr of incubation. The oviducts and uteri of 9 anestrous and 18 estrous beagle bitches were removed under halothane inhalation anesthesia to measure the total GAG amounts in oviductal and uterine fluids. The lumens of the ampulla of the oviducts, isthmus of the oviducts, and the uterine horns were each flushed with 1 ml HEPES-EDTA fluid. Total GAG amounts in the flush fluids obtained were measured with a spectrophotometer. Sperm motility (51-59%), %HY (79-86%), and %AR (31-36%) in CCM supplemented with CS, HA, or HP were significantly higher after 7 hr of incubation than when incubated in CCM without GAGs (P<0.01 or 0.05). The mean total GAG amounts in the fluids from the ampulla and isthmus of the oviducts and the uterine horns in the estrous bitches were higher than in the anestrous bitches. These results indicate that GAGs in the oviductal and uterine fluids in estrous bitches are associated with in vivo sperm capacitation.
从12只比格犬采集射出的精子,在犬获能培养基(CCM)中孵育,该培养基添加了5微克/毫升硫酸软骨素A(CS)、5微克/毫升透明质酸(HA)或5微克/毫升肝素(HP),于38℃、5%二氧化碳的空气环境中孵育7小时,以研究糖胺聚糖(GAGs)对犬精子获能的影响。孵育4小时和7小时后,检测所有培养基中活动精子、超活化精子(%HY)和顶体反应精子(%AR)的百分比。在氟烷吸入麻醉下,摘除9只处于非发情期和18只处于发情期的比格母犬的输卵管和子宫,以测量输卵管和子宫液中的总GAG含量。用1毫升HEPES - EDTA液冲洗输卵管壶腹部、输卵管峡部和子宫角的管腔。用分光光度计测量所获冲洗液中的总GAG含量。添加CS、HA或HP的CCM中,孵育7小时后的精子活力(51 - 59%)、%HY(79 - 86%)和%AR(31 - 36%)显著高于未添加GAGs的CCM孵育组(P<0.01或0.05)。发情期母犬输卵管壶腹部和峡部以及子宫角的液体中平均总GAG含量高于非发情期母犬。这些结果表明,发情期母犬输卵管和子宫液中的GAGs与体内精子获能有关。