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首页> 外文期刊>Aquaculture >Extender composition, osmolality and cryoprotectant effects on the motility of sperm in the Brazilian endangered species Brycon opalinus (Characiformes)
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Extender composition, osmolality and cryoprotectant effects on the motility of sperm in the Brazilian endangered species Brycon opalinus (Characiformes)

机译：扩展剂，重量克分子渗透压浓度和防冻剂对巴西濒危物种Brycon opalinus（Characiformes）精子活力的影响

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摘要

Sperm preservation is an important tool for conservation of endangered fish species, such as the Brycon opalinus (Characiformes). An optimum medium should prevent the initiation of sperm motility during storage. The aim of this paper was to study the effects of extender composition, osmolality and cryoprotectant agent (CPA) on the motility of B. opalinus sperm after a 30-min equilibration time. Eight media were prepared by switching extender compositions (NaCl or glucose) and osmolalities (245, 285, 325 or 365mOsm/kg). These media were then divided in three aliquots and combined with two CPAs (dimethyl sulfoxide, DMSO, or methylglycol, MG) at 1.4M; the third aliquot remained without CPA (control). After dilution, all samples were observed under light microscope to confirm whether all extender-CPA combinations actually prevented the initiation of sperm motility. Then, sperm motility was triggered in NaCl 92mOsm/kg as activating agent after 0- and 30-min equilibration time at 4A degree C and the percentage of motile sperm and duration of motility were determined. All combinations of glucose or NaCl media at high osmolalities (325 and 365mOsm/kg), completely suppressed the initiation of sperm motility. Low extender osmolalities (245mOsm/kg), however, did not prevent the initiation of sperm motility and more than 50% of sperm diluted in all combinations of glucose media, NaCl-control and in NaCl-DMSO were motile. When motility was triggered after both 0- and 30-min equilibration times, more than 77% motile sperm were observed in all combinations of NaCl and glucose media, except for glucose 245-DMSO and glucose 285-DMSO. The duration of motility in sperm diluted in all media was above 40s, except for glucose 245-DMSO. Based on these findings, we can conclude that the initiation of sperm motility is triggered by low osmolality rather than the ionic composition of the surrounding medium in B. opalinus. Glucose or NaCl solutions at high osmolalities combined with either DMSO or MG prevent the initiation of sperm motility during storage. Sperm diluted in these media yields motility upon activation above 77% and that should last long enough to fertilize oocytes. These media are recommended as freezing media for future essays in cryopreservation of B. opalinus sperm.
机译：精子的保存是保护濒危鱼类如Brycon opalinus（Characiformes）的重要工具。最佳培养基应防止在储存过程中引发精子活力。本文旨在研究补充剂组成，重量克分子渗透压浓度和冷冻保护剂（CPA）对30分钟平衡后opalinus精子活力的影响。通过切换增量剂组成（NaCl或葡萄糖）和重量摩尔渗透压浓度（245、285、325或365mOsm / kg）来制备八种培养基。然后将这些培养基分成三等份，并在1.4M下与两个CPA（二甲亚砜，DMSO或甲基乙二醇，MG）合并;第三等分试样没有CPA（对照）。稀释后，在光学显微镜下观察所有样品，以确认所有增量剂-CPA组合是否确实阻止了精子运动的开始。然后，在4A的温度下经过0分钟和30分钟的平衡时间后，以活化剂NaCl 92mOsm / kg触发精子运动，并测定运动精子的百分比和运动持续时间。高渗透压（325和365mOsm / kg）下的葡萄糖或NaCl培养基的所有组合完全抑制了精子运动的开始。然而，低渗透压摩尔渗透压浓度（245mOsm / kg）不能阻止精子活力的开始，在葡萄糖培养基，NaCl对照和NaCl-DMSO的所有组合中，稀释的精子中超过50％的是能动的。当在0分钟和30分钟的平衡时间后均触发运动时，除葡萄糖245-DMSO和葡萄糖285-DMSO外，在NaCl和葡萄糖培养基的所有组合中均观察到超过77％的运动精子。除葡萄糖245-DMSO外，在所有培养基中稀释的精子的活力持续时间均在40s以上。根据这些发现，我们可以得出结论，精子能动性的启动是由低渗透压而不是由牛油菌周围培养基的离子组成引起的。高渗透压的葡萄糖或NaCl溶液与DMSO或MG结合使用可防止在储存过程中引发精子运动。在这些培养基中稀释的精子在激活时会产生超过77％的运动力，并且应该持续足够长的时间才能使卵母细胞受精。建议将这些培养基作为冷冻培养基，以供以后冻存乳白僵菌的精子使用。

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