Department of Reproduction and Clinic of Farm Animals, Wrocław University of Environmental and Life Sciences, 50-366, Wrocław, Poland.
Department of Physics and Biophysics, Wrocław University of Environmental and Life Sciences, 50-375, Wrocław, Poland.
Sci Rep. 2024 May 3;14(1):10214. doi: 10.1038/s41598-024-61006-5.
Sperm membrane composition and biophysical characteristics play a pivotal role in many physiological processes (i.e. sperm motility, capacitation, acrosome reaction and fusion with the oocyte) as well as in semen processing (e.g. cryopreservation). The aim of this study was to characterize the fatty acid content and biophysical characteristics (anisotropy, generalized polarization) of the cell membrane of domestic cat spermatozoa. Semen was collected from 34 adult male cats by urethral catheterization. After a basic semen evaluation, the fatty acid content of some of the samples (n = 11) was evaluated by gas chromatography. Samples from other individuals (n = 23) were subjected to biophysical analysis: membrane anisotropy (which is inversely proportional to membrane fluidity) and generalized polarization (describing lipid order); both measured by fluorimetry at three temperature points: 38 °C, 25 °C and 5 °C. Spermatozoa from some samples (n = 10) were cryopreserved in TRIS egg yolk-glycerol extender and underwent the same biophysical analysis after thawing. Most fatty acids in feline spermatozoa were saturated (69.76 ± 24.45%), whereas the polyunsaturated fatty acid (PUFA) content was relatively low (6.12 ± 5.80%). Lowering the temperature caused a significant decrease in membrane fluidity and an increase in generalized polarization in fresh spermatozoa, and these effects were even more pronounced following cryopreservation. Anisotropy at 38 °C in fresh samples showed strong positive correlations with viability and motility parameters after thawing. In summary, feline spermatozoa are characterized by a very low PUFA content and a low ratio of unsaturated:saturated fatty acids, which may contribute to low oxidative stress. Cryopreservation alters the structure of the sperm membrane, increasing the fluidity of the hydrophobic portion of the bilayer and the lipid order in the hydrophilic portion. Because lower membrane fluidity in fresh semen was linked with better viability and motility after cryopreservation, this parameter may be considered an important factor in determination of sperm cryoresistance.
精子膜的组成和生物物理特性在许多生理过程中起着关键作用(例如精子运动、获能、顶体反应和与卵子融合),以及在精液处理(例如冷冻保存)中也是如此。本研究旨在描述家猫精子细胞膜的脂肪酸含量和生物物理特性(各向异性、广义极化)。通过尿道导管法从 34 只成年雄性猫中收集精液。在进行基本精液评估后,通过气相色谱法评估了一些样本(n=11)的脂肪酸含量。对其他个体的样本(n=23)进行生物物理分析:细胞膜各向异性(与膜流动性成反比)和广义极化(描述脂质有序性);均通过在三个温度点(38°C、25°C 和 5°C)通过荧光法测量。一些样本(n=10)的精子冷冻保存在 TRIS 卵黄甘油 extender 中,并在解冻后进行相同的生物物理分析。猫精子中的大多数脂肪酸都是饱和的(69.76±24.45%),而多不饱和脂肪酸(PUFA)的含量相对较低(6.12±5.80%)。降低温度会导致新鲜精子的膜流动性显著降低和广义极化增加,冷冻保存后这种影响更为明显。新鲜样本在 38°C 时的各向异性与解冻后的活力和运动参数呈强烈正相关。总之,猫精子的特点是极低的 PUFA 含量和不饱和:饱和脂肪酸的比例低,这可能有助于降低氧化应激。冷冻保存会改变精子膜的结构,增加双层疏水区的流动性和亲水区的脂质有序性。由于新鲜精液中较低的膜流动性与冷冻保存后的活力和运动能力较好相关,因此该参数可被视为确定精子抗冷冻能力的重要因素。