Kornovski B S, McCoshen J, Kredentser J, Turley E
Department of Pediatrics, Manitoba Institute of Cell Biology, Winnipeg, Canada.
Fertil Steril. 1994 May;61(5):935-40. doi: 10.1016/s0015-0282(16)56709-0.
To determine if a novel receptor for hyaluronan, termed RHAMM, is responsible for the previously observed increase in sperm locomotion in response to hyaluronan and to assess whether expression of the RHAMM protein is involved in sperm motility.
The RHAMM protein was localized on human sperm by immunofluorescence of fixed cells, fluorescence-activated cell sorter (FACS) of cell surface phenotype, and Western transblot analysis of cell proteins. The effect of monospecific antibodies on sperm motility was examined using computer-assisted image analysis. Results of motility studies were assessed statistically with analysis of variance.
Samples were collected from donors from the University of Manitoba donor insemination program.
Semen was collected twice from four participants and a total of 10,000 sperm per sample were evaluated.
A hyaluronan receptor, RHAMM, was localized by immunofluorescence along the tail, the midpiece, and the head of sperm. Positive staining obtained with FACS analysis indicated that RHAMM occurred on the surface of sperm, whereas immunoblot analysis of sperm cell lysates revealed RHAMM proteins of MWE 58 and 64 kd, consistent with the size of RHAMM localized from fibroblasts. A polyclonal antibody specific to a peptide encoded in the fibroblast RHAMM complementary DNA significantly decreased the motility of sperm. Analysis of this inhibition is consistent with an effect of the antibody on flagellar function.
The presence of RHAMM on sperm surfaces and the ability of monospecific antibodies to inhibit sperm motility suggest an important role for this novel glycoprotein in sperm motility.
确定一种名为透明质酸受体介导蛋白（RHAMM）的新型透明质酸受体是否与先前观察到的精子对透明质酸反应时运动能力增强有关，并评估RHAMM蛋白的表达是否参与精子运动。
通过固定细胞的免疫荧光、细胞表面表型的荧光激活细胞分选（FACS）以及细胞蛋白的蛋白质免疫印迹分析，将RHAMM蛋白定位在人类精子上。使用计算机辅助图像分析检查单特异性抗体对精子运动的影响。运动研究结果采用方差分析进行统计学评估。
样本取自曼尼托巴大学供精人工授精项目的捐赠者。
从四名参与者中采集两次精液，每个样本共评估10000个精子。
通过免疫荧光法将透明质酸受体RHAMM定位在精子的尾部、中段和头部。FACS分析获得的阳性染色表明RHAMM存在于精子表面，而精子细胞裂解物的免疫印迹分析显示分子量为58和64kd的RHAMM蛋白，与成纤维细胞中定位的RHAMM大小一致。一种针对成纤维细胞RHAMM互补DNA中编码肽的多克隆抗体显著降低了精子的运动能力。对这种抑制作用的分析与抗体对鞭毛功能的影响一致。
精子表面存在RHAMM以及单特异性抗体抑制精子运动的能力表明这种新型糖蛋白在精子运动中起重要作用。
