al-Anzi B, Chandler D E
Department of Biology, Arizona State University, Tempe 85287-1501, USA.
Dev Biol. 1998 Jun 15;198(2):366-75.
Although the jelly layers surrounding amphibian eggs are known to be essential to fertilization, no biological activities have been reported for jelly macromolecules in any vertebrate. Here we provide evidence that the jelly surrounding Xenopus laevis eggs releases a small diffusible protein into the surrounding media that serve as sperm chemoattractant. Using video microscopy we find that Xenopus sperm will preferentially turn toward and contact a glass capillary filled with egg jelly extract. In experiments using a two-chamber bioassay device we find that egg jelly extracts are capable of stimulating sperm movement across a membrane barrier sixfold over controls. This activity is not observed in materials unrelated to egg jelly and the response of sperm to egg jelly extract is clearly chemotactic rather than chemokinetic. A concentration gradient of the extract is absolutely necessary, and the chemotactic activity of the extract exhibits a biphasic dose dependence similar to that of chemotactic agents in other systems. We have been able to characterize the factor as being a heat-stable protein about 10 kDa in size. This study, therefore, provides the first clear evidence for a diffusible sperm chemoattractant in a nonhuman vertebrate, as well as the first demonstration of a physiological role for egg jelly macromolecules in Xenopus fertilization.
虽然已知两栖动物卵周围的胶状层对受精至关重要,但尚未有关于任何脊椎动物中胶状大分子生物活性的报道。在此,我们提供证据表明非洲爪蟾卵周围的胶状物会向周围介质释放一种可扩散的小蛋白质,该蛋白质可作为精子的化学引诱剂。通过视频显微镜观察,我们发现非洲爪蟾精子会优先转向并接触充满卵胶提取物的玻璃毛细管。在使用双室生物测定装置的实验中,我们发现卵胶提取物能够刺激精子穿过膜屏障,其运动速度比对照组快六倍。在与卵胶无关的物质中未观察到这种活性,并且精子对卵胶提取物的反应显然是趋化性的,而非化学动力学性的。提取物的浓度梯度是绝对必要的,并且提取物的趋化活性呈现出双相剂量依赖性,类似于其他系统中的趋化剂。我们已能够将该因子鉴定为一种大小约为10 kDa的热稳定蛋白质。因此,本研究首次明确证明了非人类脊椎动物中存在可扩散的精子化学引诱剂,同时也首次证明了卵胶大分子在非洲爪蟾受精过程中的生理作用。