Silla Aimee J, Keogh Leesa M, Byrne Phillip G
Centre for Sustainable Ecosystem Solutions, School of Biological Sciences, University of Wollongong, Northfields Ave, Wollongong, NSW 2522, Australia.
Reprod Fertil Dev. 2017 Oct;29(11):2277-2283. doi: 10.1071/RD17012.
Effective activation of sperm motility is fundamental to successful artificial fertilisation; however, studies investigating optimal procedures in amphibians are lacking. This study found the optimal osmolality of activation media for sperm motility activation and evaluated the effect of phosphodiesterase (PDE) inhibitors on sperm activation and longevity in the critically endangered booroolong frog, Litoria booroolongensis. To assess the effect of medium osmolality (10, 25, 50, 75, 100 and 200mOsmolkg) and PDE inhibitors (control, 2.5mM caffeine, 5mM caffeine, 2.5mM pentoxifylline, 5mM pentoxifylline, 2.5mM theophylline and 5mM theophylline) on initial activation, percentage sperm motility and sperm velocity were quantified using computer-assisted sperm analysis. To assess the effect of PDE inhibitors (control, 2.5mM caffeine and 2.5mM theophylline) on sperm longevity, percentage motility and velocity were assessed hourly until 10h after activation. High (>60%) percentage motility was achieved in a broad range of activation-medium osmolalities (10-75mOsmolkg). PDE inhibitors did not have an effect on initial sperm motility or velocity, but caffeine and theophylline improved sperm longevity, significantly increasing motility and velocity at 8, 9 and 10h after activation. Data also show that sperm longevity in L. booroolongensis is extreme, with spermatozoa remaining motile more than twice as long as those of any other anuran amphibian.
有效激活精子活力是成功进行人工授精的基础；然而，目前缺乏对两栖动物最佳操作程序的研究。本研究发现了用于激活精子活力的激活培养基的最佳渗透压，并评估了磷酸二酯酶（PDE）抑制剂对极度濒危的布罗龙蛙（Litoria booroolongensis）精子激活和寿命的影响。为了评估培养基渗透压（10、25、50、75、100和200mOsmol/kg）和PDE抑制剂（对照、2.5mM咖啡因、5mM咖啡因、2.5mM己酮可可碱、5mM己酮可可碱、2.5mM茶碱和5mM茶碱）对初始激活的影响，使用计算机辅助精子分析对精子活力百分比和精子速度进行了量化。为了评估PDE抑制剂（对照、2.5mM咖啡因和2.5mM茶碱）对精子寿命的影响，在激活后每小时评估一次活力百分比和速度，直至激活后10小时。在广泛的激活培养基渗透压范围（10 - 75mOsmol/kg）内均实现了高（>60%）活力百分比。PDE抑制剂对精子初始活力或速度没有影响，但咖啡因和茶碱提高了精子寿命，在激活后8、9和10小时显著提高了活力和速度。数据还表明，布罗龙蛙的精子寿命极长，精子保持活力的时间是其他无尾两栖动物的两倍多。
