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本文首次报道人精子甘露糖受体(MR)的电镜标记技术和初步观察。采用甘露糖化牛血清白蛋白(DMA,Sigma,A8303)结合10nm胶体金制备探针,标记获能前后精子和甲醇处理或A23187诱导顶体反应(AR)的精子。结果表明,甲醇处理后MR表达于顶体区质膜表面,活精子在AR过程中以不同方式表达MR活性;MR定位于:1.AR早期的精子顶体区质膜表面;2.AR期间精子的顶体小泡表面、顶体内膜及顶体基质;3.AR后精子的赤道带顶体内膜和顶体后区质膜;4.AR后精子的顶体后区质膜。作者认为MR原来存在于顶体内,大部分可能是膜结合蛋白。在精子发生AR的过程中MR才得以暴露并于AR后逐渐转移至赤道带和顶体后区,可能在精-卵膜融合中发挥介导作用。
This article for the first time reported human sperm mannose receptor (MR) electron microscopy and preliminary observation. Probes were prepared using mannosylated bovine serum albumin (DMA, Sigma, A8303) in combination with 10 nm colloidal gold, and sperm were tested for sperm and methanol treatment before or after stimulation or acrosome reaction (AR) induced by A23187. The results showed that after methanol treatment, MR was expressed on the surface of acrosome membrane, and live spermatozoa expressed MR activity in different ways during AR; AR early sperm acrosomal plasma membrane surface; 2. Acrosome sperm acrosomal vesicular surface, endoplasmic reticulum and acrosome during AR; 3. AR after sperm equatorial apical intima and acrosomal posterior parietal plasma membrane; 4. AR post-acrosomal sperm plasma membrane. The authors believe that MR originally exists in the acrosome, most likely membrane-bound proteins. During spermatogenesis, MR was exposed and gradually transferred to the equatorial zone and the acrosome region after AR, which may play a mediating role in sperm-egg fusion.