Gabriel L K, Franken D R, van der Horst G, Kruger T F
Department of Physiological Sciences, University of the Western Cape, Bellville, South Africa.
Arch Androl. 1994 Sep-Oct;33(2):77-85. doi: 10.3109/01485019408987807.
Wheat germ agglutinin (WGA) and Concanavalin A (Con A) seem to be the most widely studied of the lectins used to investigate sperm surface receptors. It has been reported that WGA receptors on human sperm membranes may be closely related to male fertility and that WGA receptor deficiency may cause human infertility. Although these claims may not be negated, there are many discrepancies in current literature regarding the localization and detection of human sperm WGA receptors. These discrepancies are probably the result of variations in the pretreatment of sperm before fixation and sperm-lectin interaction. This study investigated the effect of different fixation procedures on the localization of human sperm WGA receptors on normal spermatozoa. Comparative studies were performed on spermatozoa separated by wash-swim-up and fixed with 4% formaldehyde, 95% ethanol, or Bouin's solution, or nonfixed (air-dried) using fluorescein isothiocyanate-conjugated WGA. There were significant differences (p < .01) in the % WGA receptor localization following the different fixation procedures on samples receiving the same treatment previous to fixation. The different fixatives affect the plasma membrane to different degrees, resulting in an alteration of lectin receptor localization. To allow a better comparison and understanding of results, a standardized preparation procedure is recommended during studies using lectins as molecular probes. The results suggest than an aldehyde fixative may be the fixative of choice for sperm membrane studies.
小麦胚凝集素（WGA）和伴刀豆球蛋白A（Con A）似乎是用于研究精子表面受体的凝集素中研究最为广泛的。据报道，人精子膜上的WGA受体可能与男性生育能力密切相关，且WGA受体缺乏可能导致人类不育。尽管这些说法可能无法被否定，但当前文献中关于人精子WGA受体的定位和检测存在许多差异。这些差异可能是由于固定前精子预处理以及精子与凝集素相互作用的变化所致。本研究调查了不同固定程序对正常精子上人精子WGA受体定位的影响。使用异硫氰酸荧光素偶联的WGA，对通过洗涤-上游法分离并用4%甲醛、95%乙醇或布因氏液固定的精子，或未固定（风干）的精子进行了比较研究。在固定前接受相同处理的样本上，不同固定程序后的WGA受体定位百分比存在显著差异（p <.01）。不同的固定剂对质膜的影响程度不同，导致凝集素受体定位发生改变。为了更好地比较和理解结果，建议在使用凝集素作为分子探针的研究中采用标准化的制备程序。结果表明，醛类固定剂可能是精子膜研究的首选固定剂。
