Cross N L, Overstreet J W
Department of Obstetrics and Gynecology, School of Medicine, University of California, Davis 95616.
Gamete Res. 1987 Jan;16(1):23-35. doi: 10.1002/mrd.1120160104.
We have studied changes in the binding of fluoresceinated lectins to human sperm during in vitro capacitation. We first determined the surface labeling pattern of viable sperm obtained by the swim-up procedure. Sperm were labeled with 100 micrograms/ml FITC-conjugated lectin at 4 degrees C for 30 min. We simultaneously used Hoechst stain 33258 as a supravital stain to help differentiate surface from intracellular lectin labeling. Of 14 lectins studied, six (phytohemagglutinin-E, concanavalin A, Ricinus communis agglutinin-I, and the lectins of wheat germ, Lens culinaris, and Pisum sativum) bound to the entire surface of sperm, sometimes with minor local heterogeneity. Three lectins (from peanut, Maclura pomifera, and soybean) usually bound in a punctate manner, with more label on the tail than on the head. Five lectins (Ulex europaeus, Dolichos biflorus, Helix pomatia, and Vicia villosa lectins, and lectin II of Griffonia simplicifolia) bound very poorly or not at all to the sperm surface. Sperm were also inspected for changes in surface lectin binding patterns after 0, 5, and 23 hr of incubation in a capacitating medium. Two lectins showed reproducible changes. The labeling by Maclura pomifera agglutinin decreased by 5 hr in eight of ten experiments, and among sperm labeled with concanavalin A, the incidence of sperm with a highly fluorescent anterior margin of the sperm head increased by about 3.5-fold between 0 and 5 hr. The labeling pattern of the other lectins did not change.
我们研究了体外获能过程中荧光素化凝集素与人类精子结合情况的变化。我们首先确定了通过上浮法获得的活精子的表面标记模式。精子在4℃下用100微克/毫升异硫氰酸荧光素（FITC）偶联的凝集素标记30分钟。我们同时使用 Hoechst 33258 染色作为超活染色，以帮助区分表面和细胞内凝集素标记。在所研究的14种凝集素中，六种（植物血凝素-E、伴刀豆球蛋白A、蓖麻凝集素-I以及小麦胚、小扁豆和豌豆的凝集素）结合在精子的整个表面，有时存在轻微的局部异质性。三种凝集素（来自花生、桑橙和大豆）通常以点状方式结合，尾部的标记比头部更多。五种凝集素（欧洲荆豆、双花扁豆、苹果螺和绒毛野豌豆凝集素以及单叶豆凝集素II）与精子表面的结合非常差或根本不结合。还检查了精子在获能培养基中孵育0、5和23小时后表面凝集素结合模式的变化。两种凝集素显示出可重复的变化。在十次实验中的八次中，桑橙凝集素的标记在5小时时减少，在用伴刀豆球蛋白A标记的精子中，精子头部高度荧光前缘的精子发生率在0至5小时之间增加了约3.5倍。其他凝集素的标记模式没有变化。
