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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 背景：关于冷冻对射出精子和附睾精子的影响比较的实验报道很少。在目前的研究中，我们报告了在冷冻保护剂甘油存在和不存在的情况下冷却射精和附睾猴(猕猴)精子的效果。方法：用能量计以5℃/min或20℃/min的降温速率，在两种不同的冷冻条件下：(1)不加任何冷冻保护剂CPA；(2)在0.7M甘油存在的条件下，获得精液和附睾液冷冻过程中水的传输数据。根据之前公布的数值，猕猴精子细胞被建模为一个长73.83？m，半径0.16？m的圆柱体，其渗透不活跃细胞体积Vb为0.772Vo，其中Vo是等渗细胞体积。通过分析恒河猴精子的负温水运移响应来确定控制恒河猴精子冷却过程中失水率的变量，即膜通透性参数(参考膜透过率、LPG和激活能，ELP)。结果：在相应的冷却条件下，射出精子和附精精子的负温水运移反应以及相应的负温水运移参数没有显着差异。结论：如果这一观察结果被发现更普遍地适用于其他哺乳动物物种，那么在动物死后从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处理。

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.