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

在细胞外冰存在的情况下，猕猴精子在零下温度下的渗透性

• 批准号: 7958672
• 负责人: Hans Michael Kubisch
• 金额: $ 5.8万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7958672
• 关键词: Area; Cell Membrane Permeability; Cell Volumes; Computer Retrieval of Information on Scientific Projects Database; Cryoprotective Agents; Data; Equation; Freezing; Funding; Glycerol; Grant; Ice; Institution; Isotonic Exercise; Length; Macaca; Macaca mulatta; Methods; Modeling; Mole the mammal; Permeability; Primates; Publishing; Radial; Reporting; Research; Research Personnel; Resources; Source; Surface; Suspension substance; Suspensions; Translating; United States National Institutes of Health; Water; response; sperm cell

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: 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 sperm cell suspensions were obtained at a cooling rate of 20 ¿C/min in the absence of any cryoprotective agents and in the presence of 0.7 M of glycerol, as well. Using previously published values, the macaque sperm cell was modeled as a cylinder of length 73.83 ¿m with a radius of 0.40 ¿m and an osmotically inactive cell volume, Vb, of 0.772Vo, where Vo is the isotonic cell volume. This translated to a surface area, SA to initial water volume, WV ratio of ~ 22 ¿m-1. Results/Discussion: By fitting a model of water transport to the experimentally determined volumetric shrinkage data, the best-fit membrane permeability parameters (reference membrane permeability to water at 0 ¿C, Lpg or Lpg[cpa] and the activation energy, ELp or ELp[cpa]) were found to range from: Lpg or Lpg[cpa] = 0.00200.0029 ¿m/min-atm; ELp or ELp[cpa]) = 10.618.3 kcal/mole. By incorporating these membrane permeability parameters in a recently developed equation (optimal cooling rate, Bopt ¿ 1009:5 x exp(-0.0546 x ELp) x (Lpg) x (SA/WV); where the units of Bopt are ¿C/min, ELp or ELp[cpa] are kcal/mole, Lpg or Lpg[cpa] are ¿m/min-atm and SA/WV are ¿m-1), we determined the optimal rates of freezing macaque sperm to be ~ 23¿C/min (ejaculated sperm in the absence of CPAs), ~29 ¿C/min (ejaculated sperm in the presence of glycerol), ~24 ¿C/min (epididymal sperm in the absence of CPAs) and ~ 24 ¿C/min (epididymal sperm in the presence of glycerol). In conclusion, the subzero water transport response and consequently the subzero water transport parameters are not significantly different between the ejaculated and epididymal spermatozoa under corresponding cooling conditions.

这个子项目是许多研究子项目中的一个 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。 背景资料：在本研究中，我们报告的影响冷却射精和附睾恒河猴（猕猴）精子和不存在的冷冻保护剂，甘油。研究方法：在不存在任何冷冻保护剂和存在0.7 M甘油的情况下，以20 ℃/min的冷却速率获得射精和附睾精子细胞悬浮液冷冻期间的水转运数据。使用先前发表的值，猕猴精子细胞被建模为长度为73.83 μ m的圆柱体，半径为0.40 μ m，无活性细胞体积Vb为0.772 Vo，其中Vo是等渗细胞体积。这转化为表面积，SA与初始水体积，WV比约为22 μ m-1。结果/讨论：通过将水传输模型拟合到实验确定的体积收缩数据，发现最佳拟合的膜渗透性参数（0 ℃时参考膜对水的渗透性，Lpg或Lpg[cpa]和活化能，ELp或ELp[cpa]）的范围为：Lpg或Lpg[cpa] = 0.00200.0029 <$m/min-atm; ELp或ELp[cpa]）= 10.618.3千卡/摩尔。通过将这些膜渗透性参数纳入最近开发的方程，（最佳冷却速率，Bopt <$1009：5 × exp（-0.0546 × ELp）×（Lpg）×（SA/WV）;其中Bopt的单位为<$C/min，ELp或ELp[cpa]为kcal/mole，Lpg或Lpg[cpa]为<$m/min-atm，SA/WV为<$m-1），我们确定猕猴精子冷冻的最佳速率为~ 23 <$m。C/min（不含CPA的射精精子）、~29 <$C/min（含甘油的射精精子）、~24 <$C/min（不含CPA的附睾精子）和~ 24 <$C/min（含甘油的附睾精子）。总之，在相应的冷却条件下，射出精子和附睾精子之间的零度以下水运输反应和因此的零度以下水运输参数没有显著差异。