Cryopreservation effects on sperm function and fertility in two threatened crane species
Cryopreservation effects on sperm function and fertility in two threatened crane species
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DOI：
10.1016/j.cryobiol.2018.01.010
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发表时间：
2018-06-01
期刊：
影响因子：
2.7
通讯作者：
Songsasen, Nucharin
中科院分区：
文献类型：
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作者：
Brown, Megan E.;Singh, Ram P.;Songsasen, Nucharin
冷冻保存圈养鹤精液的能力有助于从行为不相容或地理上分离的对繁殖后代，并允许长期保存有价值的遗传物质。本研究旨在通过研究两种渗透性（DMA和Me2SO）和一种非渗透性（蔗糖）冷冻保护剂以及维生素E对解冻后精子存活的影响，为百日鹤（Grus americana）和白枕鹤（Grus vipio）制定有效的冷冻保存方案。在研究1中，收集射精（百日咳：n = 10，白颈：n = 8），采用两步冷却方法，在六种冷冻稀释剂（起重机扩展剂：DMA； DMA + 0.1M蔗糖；Me2SO; Me2SO + 0.1M蔗糖；0.1M蔗糖；0.2M蔗糖）中的一种中冷冻保存。冷冻样品解冻并评估其整体运动性、运动特性、膜完整性、形态和与卵泡周内膜（IPVM）结合的能力。在研究2中，将17只百日鹤的精液冷冻在单独含有Me2SO或含有维生素E （5 pg/mL或10 pg/mL）的百日鹤增液剂中。除结合试验外，冷冻样品解冻并与研究1中一样进行评估。白枕鹤精子对低温保存的耐受性高于白枕鹤（解冻后活力为15%比6%）。在这两个物种中，精子在单独含有Me2SO的培养基中冷冻保存，解冻后存活率和与IPVM结合的能力都高于其他冷冻稀释剂处理。补充维生素E对解冻后的运动或膜完整性没有好处。结果表明，对低温保存的敏感性存在物种特异性。然而，Me2SO是白枕鹤和白枕鹤精子的首选冷冻保护剂。
The capacity to cryopreserve semen from captive cranes facilitates production of offspring from behaviorally incompatible or geographically separated pairs, and allows for long-term preservation of valuable genetic materials. The present study sought to develop effective cryopreservation protocols for whooping (Grus americana) and white-naped (Grus vipio) cranes, through examining the influences of two permeating (DMA and Me2SO) and one non-permeating (sucrose) cryoprotectants, as well as vitamin E on post-thaw sperm survival. In Study 1, ejaculates (whooping: n = 10, white-naped: n = 8) were collected and cryopreserved in one of six cryo-diluents (crane extender with: DMA; DMA + 0.1M sucrose; Me2SO; Me2SO + 0.1M sucrose; 0.1M sucrose; 0.2M sucrose) using a two-step cooling method. Frozen samples were thawed and assessed for overall motility, motion characteristics, membrane integrity, morphology, and ability to bind to the inner perivitelline membrane (IPVM). In Study 2, whooping crane ejaculates (n = 17) were frozen in crane extender containing Me2SO alone or with vitamin E (5 pg/mL or 10 pg/mL). Frozen samples were thawed and assessed as in Study 1, except the binding assay. White-naped crane sperm were more tolerant to cryopreservation than whooping crane (15% vs 6% post thawed motility). In both species, sperm cryopreserved in medium containing Me2SO alone displayed higher post thaw survival and ability to bind to IPVM than the other cryodiluent treatments. Vitamin E supplementation exerted no benefits to post thaw motility or membrane integrity. The findings demonstrated that there was species specificity in the susceptibility to cryopreservation. Nevertheless, Me2SO was a preferred cryoprotectant for sperm from both whooping and white-naped cranes.