小鼠卵母细胞减数分裂过程中新型微管蛋白ε-tubulin稳定表达并与纺锤体的组装和维持相关

Stable expression as well as its correlation with spindle formation and maintenance of novel tubulin member ε-tubulin in mouse oocyte meiosis

目的研究新型微管蛋白ε-tubulin在小鼠卵母细胞减数分裂进程中的蛋白表达、亚细胞定位及其与纺锤体的相关性。方法用Western blot法检测ε-tubulin在小鼠卵母细胞减数分裂各个时期的蛋白表达;免疫荧光染色分析ε-tubulin在减数分裂进程中的亚细胞定位模式及其与纺锤体的相关性。结果ε-tubulin在小鼠卵母细胞减数分裂各个时期均有稳定的蛋白表达,在减数分裂前期均匀分布在细胞核中,在生发泡破裂后直到第二次减数分裂中期始终保持与纺锤体微管的共定位特性,并持续存在于染色体上。ε-tubulin对纺锤体毒性药物Taxol和Nocodazole敏感并呈现出与微管一致的反应性。结论小鼠卵母细胞内新型微管蛋白ε-tubulin稳定表达并与纺锤体的组装和维持相关。

Objective To investigate the protein expression and subcelluar localization of novel tubulin member ε-tubulin, as well as its relationship with spindle in mouse oocytes during meiotic maturation. Methods ε-tubulin protein expression was evaluated with Western blot, its subcelluar localization and its involvement in spindle formation was detected using immunofluorescence in mouse oocytes at different meiotic stages. Results ε-tubulin protein was stably expressed throughout the meiotic progression. ε-tubulin was evenly distributed in germinal vesicle(GV) at meiotic prophase(GV stage), and immediately relocated upon germinal vesicle breakdown(GVBD), precisely colocalized with microtubule and concentrated on chromosomes until to metaphase Ⅱ(MⅡ) stage. ε-tubulin was sensitive to spindle poisons nocodazole and taxol, showing the same reaction as spindle microtubules. ConclusionsNovel tubulin member ε-tubulin is stably expressed in mouse oocyte meiosis, and is associated with spindle formation and maintenance.