Pretreatment of Asian elephant (Elephas maximus) spermatozoa with cholesterol-loaded cyclodextrins and glycerol addition at 4°C improves cryosurvival.
Pretreatment of Asian elephant (Elephas maximus) spermatozoa with cholesterol-loaded cyclodextrins and glycerol addition at 4°C improves cryosurvival.
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DOI：
10.1071/rd11266
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Pukazhenthi BS
中科院分区：
文献类型：
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作者：
Kiso WK;Asano A;Travis AJ;Schmitt DL;Brown JL;Pukazhenthi BS
亚洲象精子对冷冻敏感，对冷冻保存反应不佳。本研究的目的是：（1）确定胆固醇含量是否可以通过用载胆固醇环糊精（CLC）预孵育亚洲象精子来改变;（2）评估CLC浓度、添加甘油时的温度（22°C vs 4°C）和稀释培养基对解冻后精子存活率的影响。精子经≥ 1.5mgCLC孵育后，胆固醇浓度显著升高（P < 0.05）。用1.5 mg CLC预处理精子，解冻后各项参数均有所改善（P < 0.05）。与22°C相比，在4°C下添加甘油也改善了所有解冻后参数。与Ham's F-10培养液相比，解冻后的精子在蛋黄培养基中的稀释度提高了精子的活力（P < 0.05）。总之，我们的研究结果表明，修改质膜内的胆固醇含量提高亚洲象精子的冷冻存活。正如本研究所报告的那样，开发一种改进的冷冻保存方法，包括修饰膜胆固醇和在4°C下添加甘油，这是在该物种的圈养管理中利用冷冻保存精子的重要一步。
Asian elephant spermatozoa are sensitive to chilling and do not respond well to cryopreservation. The objectives of the present study were to: (1) determine whether cholesterol content can be modified by preincubation of Asian elephant spermatozoa with cholesterol-loaded cyclodextrin (CLC); and (2) assess the effects of CLC concentration(s), temperature at time of glycerol addition (22°C vs 4°C) and dilution medium on post-thaw sperm survival. Spermatozoa incubated with ≥1.5 mg CLC exhibited increased (P < 0.05) cholesterol concentrations. Pretreatment of spermatozoa with 1.5 mg CLC resulted in improvements (P < 0.05) in all post-thaw parameters. Glycerol addition at 4°C also improved all post-thaw parameters compared with 22°C. Dilution of thawed spermatozoa in an egg yolk-based medium improved (P < 0.05) motility compared with Ham's F-10 culture medium. In summary, our findings indicate that modifying cholesterol content within the plasma membrane improves the cryosurvival of Asian elephant spermatozoa. The development of an improved cryopreservation method that includes modification of membrane cholesterol and the addition of glycerol at 4°C, as reported in the present study, is an important step towards utilisation of cryopreserved spermatozoa in captive management of this species.