Role of zebrafish POU-domain gene, ZFPOU2, in early development

斑马鱼 POU 结构域基因 ZFPOU2 在早期发育中的作用

• 批准号: 05680648
• 负责人: TAKEDA Hiroyuki
• 金额: $ 1.28万
• 依托单位: Nagoya University (1994-1995)The Institute of Physical and Chemical Research (1993)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05680648/
• 关键词: POU transcription factor; Zebrafish; Gastrulation; Mesoderm induction; ES cell; ES 細胞; oct-3; POU; 発生学; 分子生物学; POUファミリー; Oct-3

POU-domain proteins are a large family of transcriptional regulatory proteins, many of which are implicated in the control of gene expression during early development. We describe here the cloning and expression of zebrafish pou2, a novel POU-domain gene related to the mouse germ line-specific transcription factor Oct-3. Zebrafich pou2 is maternally expressed and the transcripts are present from the one-cell stage to the gastruls stage. In situ hybridization analyzes revealed that the transcripts were present in all blastomeres until the midblastula stage and that the expression was restricted to the epiblast during gastrulation. We found that alternatively spliced transcripts, t-pou2 RNA,were also expressed in the embryos. In contrast to the Pou2 product, t-Pou2 product lacks DNA-binding activity because of its incomplete POU-domain structure. To examine the roles of the Pou2 and t-Pou2 products, we increased their expression in the embryo by microinjection of synthetic pou2 and t-pou … More 2 RNAs into the fertilized eggs at the one-cell stage. Most embryos that developed from the eggs injected with pou2 RNA did not show any obvious developmental defects. In contrast, overexpression of the t-Pou2 product greatly affected the embryonic development : there was strong developmental retardation or arrest due to the incomplete gastrulation. In the affected embryos, expression of zebrafish T gene was reduced and the hypoblast formation was disturbed. Temporal and spatial expression patterns and the effects of overxpression of these products on development are consistent with the idea that the Pou2 and t-Pou2 proteins are involved in early development of zebrafish embryos. They may be involved in the proliferation of blastomeres in undetermined state at the blastula stage and/or the early cell commitment events at the gastrula stage. Also, our results indicate that different products generated as a result of alternative splicing from the same gene possess distinct functional capacities. Less

POU结构域蛋白是一个转录调控蛋白大家族，其中许多参与了早期发育过程中基因表达的调控。我们克隆并表达了斑马鱼POU2基因，这是一个与小鼠生殖系特异性转录因子OCT-3相关的新的POU结构域基因。Zebrafich Pou2是母性表达的，转录本存在于单细胞阶段到原肠阶段。原位杂交分析表明，直到胚泡中期，转录本在所有的卵裂球中都存在，并且在原肠胚形成过程中仅在上胚层表达。我们发现，另一种剪接的转录本t-pou2RNA也在胚胎中表达。与Pou2产物相比，t-Pou2产物缺乏DNA结合活性，因为它的POU结构域结构不完整。为了研究POU2和t-POU2产物的作用，我们通过显微注射合成的POU2和t-POU…来增加它们在胚胎中的表达单细胞期有2个以上的RNA进入受精卵。注射pou2RNA的卵子发育的大多数胚胎没有表现出任何明显的发育缺陷。相反，t-Pou2产物的过度表达极大地影响了胚胎的发育：由于原肠发育不完全，出现了强烈的发育迟缓或停滞。在受影响的胚胎中，斑马鱼T基因的表达减少，低母细胞的形成受到干扰。这些产物的时空表达模式和过度表达对发育的影响与Pou2和t-Pou2蛋白参与斑马鱼胚胎早期发育的观点一致。它们可能参与囊胚期未知状态的卵裂球的增殖和/或原肠胚期的早期细胞承诺事件。此外，我们的结果表明，由同一基因选择性剪接产生的不同产物具有不同的功能能力。较少

项目成果

Hiroyuki Takeda: "A novel POU domain gene,zebrafish pou2 : expression and roles of two alternatively spliced twin products in early development" Genes & Development. 8. 45-59 (1994)

Hiroyuki Takeda：“一种新型 POU 结构域基因，斑马鱼 pou2：两个选择性剪​​接双胞胎产物在早期发育中的表达和作用”

天沼 喜美子: "ゼブラフィッシュにおける外来遺伝子導入系" 蛋白質核酸酵素. 40. 2257-2264 (1995)

Kimiko Amanuma：“斑马鱼的外源基因转移系统”蛋白质核酸酶。40。2257-2264（1995）

Toshiro Mizuno: "Mesoderm induction in zebrafish" Nature. 383. 131-132 (1996)

Toshiro Mizuno：“斑马鱼的中胚层诱导”自然。

Terumi Miyagawa: "Specification of posterior midbrain region in zebrafish neuroepithelium" GENES TO CELLS. (印刷中). (1996)

Terumi Miyakawa：“斑马鱼神经上皮后部中脑区域的规范”（GENES TO CELLS）（1996 年出版）。

武田洋幸: "コールドスプリングハーバーLab.ミーティング「Zebratlsh Development & Genetics」に参加して" 実験医学. 12. 1676-1677 (994)

Hiroyuki Takeda：“参加冷泉港实验室会议“斑马发育与遗传学””实验医学。12. 1676-1677 (994)