Chronic cyclophosphamide exposure alters the profile of rat sperm nuclear matrix proteins
Chronic cyclophosphamide exposure alters the profile of rat sperm nuclear matrix proteins
复制标题
DOI：
10.1095/biolreprod.107.060244
复制
发表时间：
2007-08-01
影响因子：
3.6
通讯作者：
Robaire, Bernard
中科院分区：
文献类型：
--
作者：
Codrington, Alexis M.;Hales, Barbara F.;Robaire, Bernard
雄性大鼠长期暴露于烷基化剂环磷酰胺（一种众所周知的雄性介导的发育毒性物质）中，会改变雄性生殖细胞以及由暴露于环磷酰胺的雄性所产生的早期植入前胚胎的基因表达。精子DNA由核基质以序列特异性的方式组织成环结构域。在体细胞中，环结构域组织参与基因调控。核基质的各种结构和功能成分是化疗药物的靶点。因此，我们假设环磷酰胺处理会改变精子核基质蛋白的表达。用生理盐水或环磷酰胺（6.0 mg kg（-1）粘土（-1））处理成年雄性大鼠4周，从附睾尾部精子中提取核基质。通过双向凝胶电泳分析蛋白质。核基质蛋白质组中鉴定的蛋白质主要参与细胞结构、转录、翻译、DNA结合、蛋白质加工、信号转导、代谢、细胞防御或解毒等。有趣的是，环磷酰胺选择性地诱导了细胞防御和解毒蛋白的许多变化，最值得注意的是，在所有已知形式的抗氧化酶谷胱甘肽过氧化物酶4中，除了未表征的54-kDa形式；在暴露于环磷酰胺的精子的核基质提取物中观察到谷胱甘肽过氧化物酶4免疫反应性的总体增加。谷胱甘肽过氧化物酶4表达的增加表明该酶在维持核基质稳定性和功能方面的作用。这些结果使我们提出，核基质成分在药物暴露下的变化是精子功能和胚胎发育改变的一个因素。
Chronic exposure of male rats to the alkylating agent cyclophosphamide, a well-known male-mediated developmental toxicant, alters gene expression in male germ cells as well as in early preimplantation embryos sired by cyclophosphamide-exposed males. Sperm DNA is organized by the nuclear matrix into loop-domains in a sequence-specific manner. in somatic cells, loop-domain organization is involved in gene regulation. Various structural and functional components of the nuclear matrix are targets for chemotherapeutic agents. Consequently, we hypothesized that cyclophosphamide treatment would alter the expression of sperm nuclear matrix proteins. Adult male rats were treated for 4 wk with saline or cyclophosphamide (6.0 mg kg(-1) clay(-1)), and the nuclear matrix was extracted from cauda epididymal sperm. Proteins were analyzed by two-dimensional gel electrophoresis. Identified proteins within the nuclear matrix proteome were mainly involved in cell structure, transcription, translation, DNA binding, protein processing, signal transduction, metabolism, cell defense, or detoxification. Interestingly, cyclophosphamide selectively induced numerous changes in cell defense and detoxification proteins, most notably, in all known forms of the antioxidant enzyme glutathione peroxidase 4, in addition to an uncharacterized 54-kDa form; an overall increase in glutathione peroxidase 4 immunoreactivity was observed in the nuclear matrix extracts from cyclophosphamide-exposed spermatozoa. An increase in glutathione peroxidase 4 expression suggests a role for this enzyme in maintaining nuclear matrix stability and function. These results led us to propose that a change in composition of the nuclear matrix in response to drug exposure was a factor in altered sperm function and embryo development.