PERMEABILITY OF MACAQUE SPERM AT SUBZERO TEMP IN PRESENCE OF EXTACELLULAR ICE

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 背景：关于冷冻对哺乳动物射精精子和附睾精子的比较影响的实验报道很少。 在本研究中，我们报告的影响冷却射精和附睾恒河猴（猕猴）精子与不存在的冷冻保护剂，甘油。研究方法：在两种不同条件下，使用量热计以5或20 ℃/min的冷却速率获得射精和附睾猕猴精子细胞悬浮液冷冻期间的水转运数据：i）不存在任何冷冻保护剂CPA;和ii）存在0.7 M甘油。 使用先前公布的值，猕猴精子细胞被建模为长度为73.83 μ m、半径为0.16 μ m的圆柱体，无活性细胞体积Vb为0.772 Vo，其中Vo是等渗细胞体积。分析了零度以下的水运输响应，以确定猕猴精子冷却过程中水分损失速率的变量，即膜渗透性参数（参考膜渗透性，Lpg和活化能，ELp）。结果如下：在相应的冷却条件下，射精和附睾猕猴精子的零度以下水运输反应和因此的零度以下水运输参数被发现没有显着差异。结论：如果发现这种观察结果对其他哺乳动物物种也更普遍有效，那么在死后从动物睾丸中提取的精子也可以使用类似于射出精子的程序进行最佳冷冻。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Background: There are very few experimental reports on the comparative effects of freezing on ejaculated and epididymal mammalian spermatozoa. In the present study, we report the effects of cooling ejaculated and epididymal rhesus monkey (macaca mulatta) sperm with and without the presence of a cryoprotective agent, glycerol. Methods: Water transport data during freezing of ejaculated and epididymal macaque sperm cell suspensions were obtained using a calorimeter at a cooling rate of either 5 or 20 ¿C/min under two different conditions: i) in the absence of any cryoprotective agents, CPAs; and ii) in the presence of 0.7 M glycerol. Using previously published values, the macaque sperm cell was modeled as a cylinder of length 73.83 ¿m and a radius of 0.16 ¿m with an osmotically inactive cell volume, Vb, of 0.772Vo, where Vo is the isotonic cell volume. The subzero water transport response was analyzed to determine the variables governing the rate of water loss during cooling of macaque spermatozoa, i.e. the membrane permeability parameters (reference membrane permeability, Lpg and activation energy, ELp). Results: The subzero water transport response and consequently the subzero water transport parameters are found to be not significantly different between the ejaculated and epididymal macaque spermatozoa under corresponding cooling conditions. Conclusions: If this observation is found to be more generally valid for other mammalian species as well, then the sperm extracted from the testicles of an animal during post-mortem can also be optimally cryopreseved using procedures similar to those derived for ejaculated sperm.