Skiba-Lahiani M, Auger J, Terribile J, Fattal E, Delattre J, Puisieux F, Jouannet P
Université de Paris XI, Chatenay-Malabry, France.
Int J Androl. 1995 Dec;18(6):287-94. doi: 10.1111/j.1365-2605.1995.tb00564.x.
The effect of co-incubating human spermatozoa with 8 mmol/L dilauroylphosphatidylcholine (PC12) liposomes containing 6 mmol/L adenosine 5'-triphosphate (LATP) was assessed by CASA and compared to that obtained with blank PC12 liposomes (LB). The aim of this study was to investigate if such treatments can improve sperm movement and sustain sperm motility over time. Significant and similar increases in straight-line velocity and linearity of sperm movement in B2 capacitating medium (both p < 0.01) were obtained with LB and LATP treatments (final concentration: 0.38 mmol/L PC12 and 0.5 mmol/L ATP) while in Tyrode's medium supplemented with 10 mg/mL BSA, these movement parameters were increased significantly only in sperm aliquots treated with LATP. Furthermore, after incubation for 0.5 h in Tyrode's, a bioluminescence assay of intracellular ATP indicated no significant change in ATP concentration for LATP-treated spermatozoa while the ATP content of control and LB-treated spermatozoa decreased significantly during the same period (both p < 0.05). The effect of liposomes on the acrosome reaction was also investigated jointly with CASA. These experiments were performed by fluorescence microscopy, using PSA-FITC and the supravital stain Hoechst 33258. After a precapacitation period of 3 h in BWW medium the spermatozoa were incubated for 1 h with LATP, LB, LB+free ATP and free ATP alone (final concentration 0.5 mmol/L ATP). Under these conditions the percentage of acrosome-reacted spermatozoa was increased similarly after LATP and LB treatments compared to control (respectively from 4.9 to 12%, p < 0.01 and 4.9 to 11.3%, p < 0.05) but the percentage of true acrosome-reacted spermatozoa, and the values for all movement characteristics (except percentage motility) were increased significantly only with LATP treatment. The results indicate the potential of PC12 vesicles for introducing highly hydrophilic compounds into spermatozoa, as well as for modulating membrane structures and functions required for fertilization.
通过计算机辅助精子分析（CASA）评估了人类精子与含有6 mmol/L腺苷5'-三磷酸（LATP）的8 mmol/L二月桂酰磷脂酰胆碱（PC12）脂质体共孵育的效果，并与空白PC12脂质体（LB）的效果进行了比较。本研究的目的是调查这种处理是否能改善精子运动并随时间维持精子活力。在B2获能培养基中，LB和LATP处理（终浓度：0.38 mmol/L PC12和0.5 mmol/L ATP）均使精子的直线速度和运动线性显著且相似地增加（均p < 0.01）；而在补充有10 mg/mL牛血清白蛋白的Tyrode培养基中，仅LATP处理的精子等分试样的这些运动参数显著增加。此外，在Tyrode培养基中孵育0.5小时后，细胞内ATP的生物发光测定表明，LATP处理的精子的ATP浓度无显著变化，而对照和LB处理的精子的ATP含量在同一时期显著下降（均p < 0.05）。还联合CASA研究了脂质体对顶体反应的影响。这些实验通过荧光显微镜进行，使用PSA-FITC和超活染色剂Hoechst 33258。在BWW培养基中进行3小时的预获能期后，精子与LATP、LB、LB +游离ATP和单独的游离ATP（终浓度0.5 mmol/L ATP）孵育1小时。在这些条件下，与对照相比，LATP和LB处理后顶体反应精子的百分比同样增加（分别从4.9%增至12%，p < 0.01和4.9%增至11.3%，p < 0.05），但仅LATP处理使真正顶体反应精子的百分比以及所有运动特征值（除活力百分比外）显著增加。结果表明PC12囊泡有潜力将高度亲水性化合物引入精子，以及调节受精所需的膜结构和功能。
