组蛋白H3Ser10磷酸化在家蚕精母细胞减数分裂中的动态分布

【目的】探究组蛋白H3Ser10磷酸化(H3Ser10ph)在家蚕Bombyx mori精母细胞减数分裂中的功能。【方法】解剖并分离家蚕4龄幼虫至蛹期精巢组织，通过丙烯酰胺凝胶包埋制备处于减数分裂不同时期的精巢组织玻片，以免疫荧光标记检测H3Ser10ph抗体在精母细胞减数分裂不同时期的定位特点。【结果】在家蚕有核精子精母细胞减数分裂过程中，组蛋白H3Ser10的磷酸化发生在粗线期染色体的特定位置，双线期H3Ser10ph信号逐渐减弱，至终变期时在染色体上完全检测不到磷酸化信号。随着细胞周期的进行，磷酸化信号又开始逐渐增强，减数第一次分裂中期时达到最高水平。当细胞进入减数第二次分裂前中期时，染色体臂上的H3Ser10ph信号消失，在靠近纺锤体微管的分裂面处有弥散的H3Ser10ph抗体的信号，减数第二次分裂末期，仅剩余非常微弱的H3Ser10ph信号残留于染色体的特定位置。在无核精子精母细胞减数分裂过程中，在中期I至末期I一直在染色体上有较均一的3Ser10ph信号，后期I时纺锤丝微管与赤道面平行。【结论】组蛋白H3Ser10磷酸化与家蚕有核精子和无核精子精母细胞减数分裂中染色质的动态变化相关。

Dynamic distribution of histone H3 Ser10 phosphorylation during meiosis in spermatocytes of the silkworm,Bombyx mori

【Aim】 To explore the function of histone H3 Ser10 phosphorylation (H3Ser10ph) during meiosis in spermatocytes of the silkworm, Bombyx mori. 【Methods】 The testis tissues of B. mori from the 4th instar larval stage to pupal stage weredissected and separated. The slides of testis tissues at different stages of meiosis in spermatocytes were prepared by encapsulation of acrylamide gel and then the localization characteristics of the H3Ser10ph antibody was detected by using immunofluorescence staining. 【Results】 The phosphorylation of histone H3 Ser10 occurred in the specific position of pachytene chromosomes during meiosis in eupyrene spermatocytes of B. mori. The signal of H3Ser10ph was gradually weakened in the diplotene, and no phosphorylation signal was detected in the chromosomes at the diakinesis. With the progress of cell cycle, phosphorylation signal began to increase, and reached the highest level at the metaphase of the first meiosis. When the cells entered the prometaphase II of meiosis, the signal of H3Ser10ph on the chromosome arm disappeared, and there was holo-shaped fluorescent signal of the H3Ser10ph antibody close to the spindle microtubulin attaching side. At the end of meiosis II, a weak H3Ser10ph signal remained in the specific position of chromatin. During apyrene spermatocyte meiosis, the signal of H3Ser10ph was aggregated uniformly along the whole chromosomes from metaphase I to telophase I, and the spindle microtubules were parallel to the equatorial plane at anaphase I. 【Conclusion】 Histone H3 Ser10 phosphorylation is correlated with the dynamic changes of chromatin in eupyrene spermatocytes and apyrene spermatocytes of the silkworm.