Blesbois E, Grasseau I, Seigneurin F
Station de Recherches Avicoles, INRA-SYSAAF, 37380 Nouzilly, France.
Reproduction. 2005 Mar;129(3):371-8. doi: 10.1530/rep.1.00454.
The ability to survive cryopreservation varies in spermatozoa from different bird species. Among the biological factors potentially responsible for such differences, species variations in membrane fluidity have a role in the restoration of the physiological state after freezing. Membrane fluidity may be assessed by measuring fluorescence polarization anisotropy with a fluorescent dye. Anistropy values are proportional to membrane rigidity and consequently inversely proportional to membrane fluidity. In the present study, polarization anisotropy of spermatozoa originating from species differing in the freezability of their semen (chicken, turkey and guinea fowl) was measured in addition to lipid composition (cholesterol/phospholipid ratio), sperm viability (membrane permeability to eosine) and morphological integrity before and after cryopreservation. The percentages of viable and normal spermatozoa in fresh sperm were highest in the chicken (87%), lowest in guinea fowl (64%), and intermediate in turkeys (69%). Anisotropy values were highest in guinea fowl (0.205), lowest in chickens (0.155), and intermediate in turkeys (0.180). As a consequence, membrane fluidity was highest in chickens and lowest in guinea fowl. Cryopreservation significantly decreased sperm viability and morphological integrity and increased anisotropy in all species but did not change the inter species hierarchy. Initial cholesterol/phospholipid ratios were lower in chickens than in guinea fowl, and intermediate in turkeys (0.25, 0.26 and 0.29, respectively). Cryopreservation induced a severe decrease in cholesterol/phospholipid ratios in turkeys and guinea fowl. Sperm membrane fluidity in chickens, turkeys and guinea fowl behaves as an indicator of sperm freezability in these species. Inter species differences for this parameter may be partly explained by differences in initial cholesterol/phospholipids content of spermatozoa. On the other hand, the rigidifying process induced by cryopreservation is not related to lipid damage by the same mechanisms.
不同鸟类物种的精子在冷冻保存后的存活能力各不相同。在可能导致这种差异的生物学因素中,膜流动性的物种差异在冷冻后生理状态的恢复中起作用。膜流动性可以通过用荧光染料测量荧光偏振各向异性来评估。各向异性值与膜刚性成正比,因此与膜流动性成反比。在本研究中,除了冷冻保存前后的脂质组成(胆固醇/磷脂比率)、精子活力(对伊红的膜通透性)和形态完整性外,还测量了来自精液冷冻能力不同的物种(鸡、火鸡和珍珠鸡)的精子的偏振各向异性。新鲜精子中活精子和正常精子的百分比在鸡中最高(87%),在珍珠鸡中最低(64%),在火鸡中居中(69%)。各向异性值在珍珠鸡中最高(0.205),在鸡中最低(0.155),在火鸡中居中(0.180)。因此,膜流动性在鸡中最高,在珍珠鸡中最低。冷冻保存显著降低了所有物种的精子活力和形态完整性,并增加了各向异性,但没有改变物种间的等级顺序。鸡的初始胆固醇/磷脂比率低于珍珠鸡,火鸡居中(分别为0.25、0.26和0.29)。冷冻保存导致火鸡和珍珠鸡的胆固醇/磷脂比率严重下降。鸡、火鸡和珍珠鸡的精子膜流动性表现为这些物种精子冷冻能力的一个指标。该参数的物种间差异可能部分由精子初始胆固醇/磷脂含量的差异来解释。另一方面,冷冻保存诱导的僵化过程与脂质损伤的机制不同。