Mizuno Masashi, Donev Rossen M, Harris Claire L, Morgan B Paul
Complement Biology Group, Department of Medical Biochemistry and Immunology, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, UK.
Mol Immunol. 2007 Mar;44(7):1613-22. doi: 10.1016/j.molimm.2006.08.018. Epub 2006 Sep 27.
CD55 is a key regulator of complement activation, expressed on most tissues and cells in man and other mammals. In the rat, alternative splicing in the gene encoding CD55 yields GPI-anchored (GPI-CD55) and transmembrane (TM-CD55) forms. Published Northern blot analysis indicated that while GPI-CD55 was broadly expressed, TM-CD55 was primarily expressed in the testis, although the precise site of expression was not identified. To clarify the distribution of CD55 isoforms in rat reproductive tissues, we first performed immunohistochemistry and Western blot analysis with an anti-rat CD55 mAb that recognized all reported CD55 isoforms, and a polyclonal immunoglobulin specific for TM-CD55. CD55 was absent in testis prior to puberty. Post-puberty, CD55 was expressed at high levels on all spermiogenic cells from step 6 spermatid onward, and on mature spermatozoa focussed on the acrosome, but was absent from support cells and early progenitors. Enzymatic digestion revealed that GPI-CD55 was predominant in testis and spermatozoa. Staining for TM-CD55 with specific immunoglobulin confirmed its absence from mature sperm and expression on spermatids only between steps 11 and 14 of development. GPI-CD55 on spermatozoa was of lower molecular weight than that in testis and other tissues; sequencing from spermatozoal mRNA identified a unique isoform of GPI-CD55 missing short consensus repeat 4. The predominant acrosome expression and presence of a unique, truncated isoform of CD55 on spermatozoa provides further support for the hypothesis that the acrosome is a highly specialized region in which closely regulated complement activation may contribute to reproductive function.
CD55是补体激活的关键调节因子,在人类和其他哺乳动物的大多数组织和细胞上表达。在大鼠中,编码CD55的基因发生可变剪接,产生糖基磷脂酰肌醇锚定(GPI-CD55)和跨膜(TM-CD55)形式。已发表的Northern印迹分析表明,虽然GPI-CD55广泛表达,但TM-CD55主要在睾丸中表达,尽管其确切表达位点尚未确定。为了阐明CD55异构体在大鼠生殖组织中的分布,我们首先用一种识别所有已报道的CD55异构体的抗大鼠CD55单克隆抗体和一种针对TM-CD55的多克隆免疫球蛋白进行免疫组织化学和蛋白质印迹分析。青春期前睾丸中不存在CD55。青春期后,从第6步精子细胞开始的所有生精细胞以及集中在顶体上的成熟精子上均高水平表达CD55,但支持细胞和早期祖细胞中不存在。酶消化显示GPI-CD55在睾丸和精子中占主导地位。用特异性免疫球蛋白对TM-CD55进行染色证实其在成熟精子中不存在,仅在发育的第11至14步之间的精子细胞上表达。精子上的GPI-CD55分子量低于睾丸和其他组织中的;从精子mRNA测序鉴定出一种缺失短共有重复序列4的独特GPI-CD55异构体。精子上主要的顶体表达和独特截短的CD55异构体的存在为顶体是一个高度特化区域的假说提供了进一步支持,在该区域中紧密调节的补体激活可能有助于生殖功能。