Harris Merrilee S, Rodger John C
School of Environmental and Life Sciences, University of Newcastle, NSW 2308, Australia.
J Exp Zool A Comp Exp Biol. 2005 Aug 1;303(8):713-21. doi: 10.1002/jez.a.193.
Monoclonal antibodies (mAb) have been raised against marsupial sperm proteins to provide insights into the molecular nature of marsupial spermatozoa, and the proteins that mediate sperm maturation and interaction with the oocyte. This study reports the production of a mAb, designated WSA-1, which bound acrosomal and surface determinants on tammar wallaby spermatozoa. The acrosomal antigen was first detected in the wallaby testis; however, ejaculated spermatozoa demonstrated whole cell WSA-1 immunoreactivity as a result of binding an epididymal protein. Ultrastructural and agglutination analyses localised the WSA-1 epitope to the acrosomal matrix and the whole sperm plasmalemma. The WSA-1 mAb bound three polypeptides with relative molecular weights of 35, 31 and 15 kDa on western blots under reducing conditions. The N-terminal amino acid sequence obtained for the 35 kDa wallaby sperm polypeptide demonstrated identity with the eutherian acrosomal protein acrosin. The 31 kDa polypeptide was of epididymal origin and will be the subject of a separate study. Further studies of the WSA-1 antigens are likely to provide useful insights into the function and maturation of marsupial sperm since proacrosin has a number of putative roles in eutherian fertilisation, and epididymal proteins are thought to mediate sperm maturation and storage.
已经制备了针对有袋动物精子蛋白的单克隆抗体（mAb），以深入了解有袋动物精子的分子性质，以及介导精子成熟和与卵母细胞相互作用的蛋白质。本研究报告了一种名为WSA-1的单克隆抗体的产生，它能与袋鼠精子的顶体和表面决定簇结合。顶体抗原最初在袋鼠睾丸中被检测到；然而，由于与附睾蛋白结合，射出的精子表现出全细胞WSA-1免疫反应性。超微结构和凝集分析将WSA-1表位定位到顶体基质和整个精子质膜上。在还原条件下，WSA-1单克隆抗体在western印迹上结合了三种相对分子质量分别为35、31和15 kDa的多肽。从35 kDa袋鼠精子多肽获得的N端氨基酸序列与真兽类顶体蛋白顶体蛋白酶相同。31 kDa多肽起源于附睾，将是另一项研究的主题。对WSA-1抗原的进一步研究可能会为有袋动物精子的功能和成熟提供有用的见解，因为前顶体蛋白酶在真兽类受精中有许多假定的作用，并且附睾蛋白被认为介导精子成熟和储存。
