A rat H1t-GFP transgene recapitulates endogenous H1t expression pattern in mouse.
A rat H1t-GFP transgene recapitulates endogenous H1t expression pattern in mouse.
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大鼠 H1t-GFP 转基因重现了小鼠内源性 H1t 表达模式。
DOI：
10.1002/dvg.23355
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Wang,PJeremy
中科院分区：
文献类型：
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作者：
Lipschutz,Emma;Dasgupta,Anindya;Guan,Yongjuan;Kistler,WStephen;Wang,PJeremy
H1 组蛋白与接头 DNA 结合。 H1t (H1f6) 是一种睾丸特异性连接组蛋白变体，存在于粗线期精母细胞和精子细胞中。 H1t 组蛋白的表达与粗线期精母细胞中期 I 能力的获得一致。在这里，我们报告了 H1t-GFP 转基因小鼠的产生。 H1t-GFP（H1 组蛋白睾丸绿色荧光蛋白）融合蛋白表达概括了内源性 H1t 表达模式。该蛋白首先出现在第五阶段生精小管的粗线期中期精母细胞中，持续存在于圆形精子细胞和伸长精子细胞中，但在伸长精子细胞中不存在。由于 H1t-GFP 丰度高，在用冈田酸处理诱导进入中期 I 后，精母细胞中仍保持强烈的绿色荧光信号。因此，H1t-GFP可用作监测体外和体内减数分裂进展以及生殖细胞荧光激活细胞分选（FACS）分选的视觉标记。
H1 histones bind to linker DNA. H1t (H1f6), a testis‐specific linker histone variant, is present in pachytene spermatocytes and spermatids. The expression of H1t histone coincides with the acquisition of metaphase I competence in pachytene spermatocytes. Here we report the generation of H1t‐GFP transgenic mice. The H1t‐GFP (H1 histone testis‐green fluorescence protein) fusion protein expression recapitulates the endogenous H1t expression pattern. This protein appears first in mid pachytene spermatocytes in stage V seminiferous tubules, persists in round spermatids and elongating spermatids, but is absent in elongated spermatids. The strong green fluorescence signal, due to the high abundance of H1t‐GFP, is maintained in spermatocytes after induction towards metaphase I through treatment with okadaic acid. Therefore, H1t‐GFP can be used as a visual marker for monitoring the progression of meiosis in vitro and in vivo, as well as fluorescence‐activated cell sorting (FACS) sorting of germ cells.