Sidhu K S, Mate K E, Rodger J C
Cooperative Research Centre for Conservation and Management of Marsupials, School of Biological Sciences, Macquarie University, NSW, Australia.
J Reprod Fertil. 1998 Sep;114(1):55-61. doi: 10.1530/jrf.0.1140055.
Oviduct epithelial cell (OEC) monolayers were prepared from the isthmic and ampullary parts of the oviducts of FSH-primed tammar wallabies. Co-culture experiments found that 50-60% of wallaby spermatozoa attached immediately to OEC monolayers, tracheal cell monolayer controls, and the surface of culture dishes with and without Matrigel coating. Spermatozoa were considered to be attached if they remained on the culture surface after rapidly pipetting the co-culture medium five times. The percentages of attached and unattached spermatozoa were calculated from the number of spermatozoa recovered in the agitated supernatant. After 2 h co-culture the percentage of attached spermatozoa rose to 60-80%. After 6 h co-culture the number of spermatozoa attached to OEC monolayers derived from the oviductal isthmus remained high and only a small percentage were recovered in the agitated supernatant (unattached spermatozoa 3.85 +/- 0.76%, P = 0.67). However, after 6 h co-culture of spermatozoa with OEC monolayers derived from the ampulla and with the controls the percentage of attached spermatozoa declined significantly (unattached spermatozoa: ampullary monolayer 23.08 +/- 4.80%, P < 0.01; tracheal monolayer 23.23 +/- 5.18%, P < 0.01; Matrigel 27.23 +/- 7.76%, P < 0.01; plastic surface 28.19 +/- 5.30%, P < 0.01). After 6 h co-culture with ampullary and isthmic OEC monolayers, the percentage motility of both attached and unattached spermatozoa was maintained at 64.00 +/- 1.90% and 56.66 +/- 3.18% and 62.00 +/- 3.11% and 52.00 +/- 2.43%, respectively, and was then maintained at > or = 35% after 24 h incubation. In the controls, that is, tracheal monolayer and Matrigel, the motility of attached spermatozoa declined rapidly to 48.66 +/- 2.15% and 33.63 +/- 8.66%, respectively, at 6 h, and all spermatozoa were immotile after 24 h incubation. However, the motility of unattached spermatozoa in the controls (tracheal monolayer and Matrigel) was maintained at 57.33 +/- 3.00% and 34.54 +/- 9.27%, respectively, until 6 h and then declined rapidly, and all spermatozoa were immotile after 24 h incubation. Co-culture of wallaby spermatozoa with OEC monolayers also induced acrosomal modifications that were followed by acrosomal loss. At 6 h incubation 38.92 +/- 3.98% of spermatozoa on ampullary OEC monolayers and 36.50 +/- 3.81% spermatozoa on isthmic OEC monolayers had shed their acrosome. Acrosomal loss during co-culture with both isthmic and ampullary OEC monolayers was significantly (P < 0.01) greater than that observed on tracheal epithelial monolayer (24.42 +/- 1.90%, P < 0.01), Matrigel (20.70 +/- 2.71%, P < 0.01) and plastic (15.54 +/- 2.49%, P < 0.01). Co-culturing spermatozoa with OEC monolayers also induced a transformation from streamlined orientation of sperm head and tail to T-shaped (thumbtack) orientation in a small number (10-15%) of motile spermatozoa after 6 h incubation (data not shown). The significance of these results in relation to the role of the oviduct in sperm capacitation is discussed.
从经促卵泡素（FSH）预处理的帚尾袋鼩输卵管峡部和壶腹部制备输卵管上皮细胞（OEC）单层。共培养实验发现，50% - 60%的袋鼩精子立即附着于OEC单层、气管细胞单层对照以及有或没有基质胶包被的培养皿表面。如果在快速吸取共培养基5次后精子仍留在培养表面，则认为精子已附着。附着和未附着精子的百分比根据搅拌上清液中回收的精子数量计算。共培养2小时后，附着精子的百分比升至60% - 80%。与来自输卵管峡部的OEC单层共培养6小时后，附着于其上的精子数量仍然很高，在搅拌上清液中回收的未附着精子比例很小（未附着精子为3.85±0.76%，P = 0.67）。然而，精子与来自壶腹部的OEC单层以及对照共培养6小时后，附着精子的百分比显著下降（未附着精子：壶腹部单层为23.08±4.80%，P < 0.01；气管单层为23.23±5.18%，P < 0.01；基质胶为27.2(3±7.76%，P < 0.01；塑料表面为28.19±5.30%，P < 0.01）。与壶腹部和峡部OEC单层共培养6小时后，附着和未附着精子的活力百分比分别维持在64.00±1.90%和56.66±3.18%以及62.00±3.11%和52.00±2.43%，然后在孵育24小时后维持在≥35%。在对照中，即气管单层和基质胶，附着精子的活力在6小时时分别迅速降至48.66±2.15%和33.63±8.66%，孵育24小时后所有精子均失去活力。然而，对照（气管单层和基质胶）中未附着精子的活力分别维持在57.33±3.00%和34.54±9.27%直至6小时，然后迅速下降，孵育24小时后所有精子均失去活力。袋鼩精子与OEC单层共培养还诱导了顶体修饰，随后顶体丢失。孵育6小时时，壶腹部OEC单层上38.92±3.98%的精子和峡部OEC单层上36.50±3.81%的精子已脱去顶体。与峡部和壶腹部OEC单层共培养期间的顶体丢失显著（P < 0.01）大于在气管上皮单层（24.42±1.90%，P < 0.01）、基质胶（20.70±2.71%，P < 0.01）和塑料（15.54±2.49%，P < 0.01）上观察到的情况。孵育6小时后，与OEC单层共培养还在少数（10% - 15%）活动精子中诱导了精子头尾流线型取向到T形（图钉形）取向的转变（数据未显示）。讨论了这些结果与输卵管在精子获能中的作用的相关性。
