Kawakami E, Kashiwagi C, Hori T, Tsutsui T
Department of Reproduction, Nippon Veterinary and Animal Science University, 1-7-1 Kyonan-cho, Musashino-shi, 180-8602, Tokyo, Japan.
Anim Reprod Sci. 2001 Oct 31;68(1-2):121-31. doi: 10.1016/s0378-4320(01)00135-x.
In this study, the interaction between canine sperms and oviduct epithelial cells (OECs) was examined in vitro. The oviducts of eight bitches in the follicular (F-) phase and six bitches in the luteal (L-) phase were removed under halothane inhalation anesthesia. The entire oviduct was opened longitudinally, and the oviductal epithelium of bitches in the F- and L-phases was scraped with a scalpel into tissue culture medium (Eagle's MEM) containing 10% estrous bitch serum and 10% diestrous bitch serum, respectively. The OEC collected were preincubated for 24h and then coincubated with ejaculated canine sperms at 38 degrees C under an atmosphere of 5% CO2 in air. The percentages of sperms exhibiting active tail movement (% TM), hyperactivated sperms (% HA), and acrosome-reacted sperms (% AR) were investigated until 72 h after the start of coincubation. The percentage of sperms labeled with fluoresceinated Ca indicator (% Ca) was evaluated to assess the influx of Ca into sperms cytoplasm during capacitation. Canine sperms attached to both ciliated OEC and non-ciliated OEC. All of the mean % TM of the OEC-binding sperms in the F-OEC and L-OEC media after 24, 48, and 72 h of coincubation were significantly higher than the values of the freely swimming sperms (P< or =0.01). Conversely, the mean % AR and % Ca of the OEC-binding sperms were significantly lower (P<0.01). All of the mean % HA and % AR of the freely swimming sperms in the F-OEC medium after 24, 48, and 72 h of coincubation were significantly higher than the values of the sperms in the L-OEC medium (P< or =0.01). These results indicate that attachment of canine sperms to the OEC prolongs their viability and motility arid inhibits Ca influx into the sperms and sperm capacitation. These phenomena may be responsible for maintaining the active movement and the fertile life of canine sperms in homologous oviducts.
在本研究中，对犬精子与输卵管上皮细胞（OECs）之间的相互作用进行了体外研究。在氟烷吸入麻醉下，摘除处于卵泡期（F期）的8只母犬和处于黄体期（L期）的6只母犬的输卵管。将整个输卵管纵向切开，用手术刀分别刮取F期和L期母犬的输卵管上皮，放入含有10%发情期母犬血清和10%间情期母犬血清的组织培养基（伊格尔氏MEM培养基）中。收集到的OECs先预孵育24小时，然后在38℃、5%二氧化碳的空气环境中与射出的犬精子共同孵育。在共同孵育开始后72小时内，研究表现出活跃尾部运动的精子百分比（%TM）、超活化精子百分比（%HA）和顶体反应精子百分比（%AR）。评估用荧光钙指示剂标记的精子百分比（%Ca），以评估获能过程中钙离子流入精子细胞质的情况。犬精子可附着于有纤毛的OEC和无纤毛的OEC。共同孵育24、48和72小时后，F期OEC培养基和L期OEC培养基中与OEC结合的精子的所有平均%TM均显著高于自由游动精子的值（P≤0.01）。相反，与OEC结合的精子的平均%AR和%Ca显著较低（P<0.01）。共同孵育24、48和72小时后，F期OEC培养基中自由游动精子的所有平均%HA和%AR均显著高于L期OEC培养基中精子的值（P≤0.01）。这些结果表明，犬精子与OEC的附着延长了其活力和运动能力，并抑制了钙离子流入精子和精子获能。这些现象可能是维持犬精子在同源输卵管中的活跃运动和受精能力的原因。
