del Priore Lucía, Pigozzi María Inés
INBIOMED, CONICET/University of Buenos Aires, Buenos Aires, Argentina.
Chromosoma. 2014 Jun;123(3):293-302. doi: 10.1007/s00412-014-0451-3. Epub 2014 Feb 4.
We report here that a germline-restricted chromosome (GRC) is regularly present in males and females of the Bengalese finch (Lonchura domestica). While the GRC is euchromatic in oocytes, in spermatocytes this chromosome is cytologically seen as entirely heterochromatic and presumably inactive. The GRC is observed in the cytoplasm of secondary spermatocytes, indicating that its elimination from the nucleus occurs during the first meiotic division. By immunofluorescence on microspreads, we investigated the presence of histone H3 modifications throughout male meiosis, as well as in postmeiotic stages. We found that the GRC is highly enriched in di- and trimethylated histone H3 at lysine 9 during prophase I, in agreement with the presumed inactive state of this chromosome. At metaphase I, dimethylated histone H3 is no longer detectable on the GRC and its chromatin is more faintly stained with DAPI. The condensed GRC is underphosphorylated at serine 10 compared to the regular chromosomes during metaphase I, being phosphorylated later at this site after the first meiotic division. From these results, we proposed that trimethylation of histone H3 at lysine 9 on the GRC chromatin increases during metaphase I. This hypermethylated state at lysine 9 may preclude the phosphorylation of the adjacent serine 10 residue, providing an example of cross-talk of histone H3 modifications as described in experimental systems. The differential underphosphorylation of the GRC chromatin before elimination is interpreted as a cytologically detectable byproduct of deficient activity of Aurora B kinase, which is responsible for the phosphorylation of H3 at serine 10 during mitosis and meiosis.
我们在此报告,在 Bengalese 雀(Lonchura domestica)的雄性和雌性个体中,常存在一种生殖系限制染色体(GRC)。虽然 GRC 在卵母细胞中是常染色质,但在精母细胞中,这条染色体在细胞学上被视为完全异染色质,推测是无活性的。在次级精母细胞的细胞质中可观察到 GRC,这表明它在第一次减数分裂期间从细胞核中被消除。通过对微铺展样本进行免疫荧光检测,我们研究了整个雄性减数分裂过程以及减数分裂后阶段中组蛋白 H3 修饰的存在情况。我们发现,在前期 I 期间,GRC 高度富集赖氨酸 9 处的二甲基化和三甲基化组蛋白 H3,这与该染色体的推测无活性状态一致。在中期 I,GRC 上不再能检测到二甲基化组蛋白 H3,其染色质用 DAPI 染色也更淡。与中期 I 的正常染色体相比,浓缩的 GRC 在丝氨酸 10 处磷酸化不足,在第一次减数分裂后该位点稍后才被磷酸化。基于这些结果,我们提出在中期 I 期间,GRC 染色质上赖氨酸 9 处的组蛋白 H3 三甲基化增加。赖氨酸 9 处的这种高甲基化状态可能会阻止相邻丝氨酸 10 残基的磷酸化,这提供了一个如实验系统中所描述的组蛋白 H3 修饰相互作用的例子。在消除之前 GRC 染色质的差异磷酸化不足被解释为 Aurora B 激酶活性不足的细胞学可检测副产物,Aurora B 激酶在有丝分裂和减数分裂期间负责 H3 在丝氨酸 10 处的磷酸化。