Gamzu R, Yogev L, Paz G, Yavetz H, Lichtenberg D
Institute for the Study of Fertility, Serlin Maternity Hospital, Tel Aviv, Israel.
Biol Reprod. 1997 Sep;57(3):539-46. doi: 10.1095/biolreprod57.3.539.
TEST (TES (N-tris[hydroxymethyl]methyl-2-aminoethanesulfonic acid) and Tris) yolk buffer (TYB) has recently been shown to improve the binding capacity of spermatozoa to zona pellucidae. The present study had two objectives: 1) to elucidate which component(s) of TYB dominates this effect and 2) to define the responsible mechanism. Sperm samples obtained from subfertile men were incubated for 2 h in either TYB or media containing egg yolk lipoproteins or phospholipids. After incubation, sperm binding was tested by the hemizona assay. Yolk lipoprotein-treated spermatozoa bound hemizonae with efficiency equal to that of the spermatozoa incubated in control medium. Conversely, incubation of spermatozoa in media containing either TYB, yolk-phospholipids, or pure phosphatidylcholine (PC) resulted in a 2- to 3-fold increased binding capacity (p < 0.01). A close correlation was found between the effect of yolk-phospholipids and TYB on the binding capacity of the same sperm samples, compared to spermatozoa incubated in control medium. Incubation of spermatozoa in yolk phospholipid medium caused a dose-dependent increase of sperm binding capacity (p < 0.05). Treatment of sperm samples with 1 mg/ml or more of purified PC preparation also resulted in a reduction of the sperm cholesterol:phospholipid molar ratio. Significant correlations between the effects of the treatments on sperm cholesterol: phospholipid molar ratio and sperm binding were obtained with yolk-phospholipids (r = -0.55) or 1 mg/ml purified PC (r = -0.61). We conclude that 1) the enhanced binding capacity of human spermatozoa following TYB treatment is probably due to yolk-phospholipids, mainly egg yolk PC; and 2) it appears that the enhanced binding capacity of human spermatozoa following treatment with egg yolk-containing media may be a result of the reduction of the cholesterol:phospholipid molar ratio in the sperm cells.
TEST(TES,N-三[羟甲基]甲基-2-氨基乙烷磺酸)和Tris蛋黄缓冲液(TYB)最近已被证明可提高精子与透明带的结合能力。本研究有两个目的:1)阐明TYB的哪些成分主导这种作用;2)确定其作用机制。从生育力低下的男性获得的精子样本在TYB或含有蛋黄脂蛋白或磷脂的培养基中孵育2小时。孵育后,通过半透明带试验检测精子结合情况。用蛋黄脂蛋白处理的精子与半透明带的结合效率与在对照培养基中孵育的精子相同。相反,将精子在含有TYB、蛋黄磷脂或纯磷脂酰胆碱(PC)的培养基中孵育,会使结合能力提高2至3倍(p<0.01)。与在对照培养基中孵育的精子相比,发现蛋黄磷脂和TYB对同一精子样本结合能力的影响之间存在密切相关性。在蛋黄磷脂培养基中孵育精子会导致精子结合能力呈剂量依赖性增加(p<0.05)。用1mg/ml或更高浓度的纯化PC制剂处理精子样本也会导致精子胆固醇:磷脂摩尔比降低。用蛋黄磷脂(r=-0.55)或1mg/ml纯化PC(r=-0.61)处理后,处理对精子胆固醇:磷脂摩尔比和精子结合的影响之间存在显著相关性。我们得出结论:1)TYB处理后人精子结合能力增强可能归因于蛋黄磷脂,主要是蛋黄PC;2)用含蛋黄培养基处理后人精子结合能力增强似乎可能是精子细胞中胆固醇:磷脂摩尔比降低的结果。