FLASH/casp8ap2 is indispensable for early embryogenesis but dispensable for proliferation and differentiation of ES cells.
FLASH/casp8ap2 is indispensable for early embryogenesis but dispensable for proliferation and differentiation of ES cells.
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DOI：
10.1371/journal.pone.0108032
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Yonehara S
中科院分区：
文献类型：
--
作者：
Minamida Y;Someda M;Yonehara S
FLICE/caspase-8相关巨大蛋白（FLASH）/caspase 8ap 2参与多种细胞功能，如细胞周期进程、转录调控、凋亡调控和组蛋白基因表达调控。在多种小鼠和人细胞系中，FLASH的表达下调可抑制细胞周期进程于S期，其抑制细胞周期进程的机制可能与抑制复制依赖性组蛋白基因的表达有关。我们在这里证明，诱导敲除FLASH从来没有影响ES细胞的细胞周期进程，其中核心组蛋白基因的表达降低到类似于人KB细胞对FLASH敲除敏感的水平。此外，FLASH条件性基因敲除ES细胞不仅可以正常分化为中胚层和内胚层细胞，而且还可以分化为滋养层细胞。为了研究FLASH在体内早期胚胎发生中的作用，我们还检测了一只FLASH突变小鼠，其中FLASH突变等位基因在胚胎和除睾丸外的大多数成年器官中不表达FLASH mRNA。FLASH突变胚胎在E3.5和E8.5之间死亡。此外，通过体外受精产生的FLASH突变体胚胎的体外培养通过抑制胚胎的孵化及其对基质的粘附而显示出在植入前阶段的胚胎致死性。综上所述，这些结果表明FLASH在早期胚胎发生中起重要作用，但对于ES细胞的增殖或分化不是必需的。
FLICE/caspase-8-associated huge protein (FLASH)/casp8ap2 is involved in various cellular functions, such as cell cycle progression, transcriptional regulation, the regulation of apoptosis, and the regulation of histone gene expression. The down-regulated expression of FLASH has been shown to inhibit cell cycle progression in the S phase in many kinds of mice and human cell lines and the inhibition of cell cycle progression may be attributed to the suppressed expression of replication-dependent histone genes. We here demonstrated that the induced knockout of FLASH never affected cell cycle progression in ES cells, in which the expression of core histone genes was decreased to levels similar to those in human KB cells sensitive to the knockdown of FLASH. In addition, the FLASH conditional knockout ES cells could differentiate normally into not only mesodermal and endodermal cells, but also trophoblasts. In order to investigate the function of FLASH in early embryogenesis in vivo, we also examined a FLASH mutant mouse, in which FLASH mutant allele did not express FLASH mRNA in embryos and most adult organs, except for the testis. FLASH mutant embryos died between E3.5 and E8.5. Furthermore, the in vitro cultivation of FLASH mutant embryos generated by in vitro fertilization showed embryonic lethality at the pre-implantation stage by inhibiting the hatching of embryos and their adherence to substrates. Taken together, these results indicate that FLASH plays an important role in early embryogenesis, but is not essential for either the proliferation or differentiation of ES cells.
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影响因子：
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