Identification and functional analysis of human oocyte-specific linker histone H1

人卵母细胞特异性接头组蛋白 H1 的鉴定和功能分析

• 批准号: 17591757
• 负责人: KUJI Naoaki
• 金额: $ 2.24万
• 依托单位: KEIO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591757/
• 关键词: oocytes; histones; H1; decondensation; 移植・再生医療; 医療・福祉; 細胞・組織; 蛋白質; 発生・分化

We investigated osH1 protein expression in the context of biologic significance in human fertilization. A polyclonal rabbit antibody raised against synthetic peptides representing parts of the osH1 sequence was used for Western blotting of extracts from oocytes and ovarian tissues, which demonstrated a band at 36kDa, the calculated molecular mass of osH1. Western analysis detected the protein in chromatin of germinal-vesicle-stage oocytes through two-cell embryos. Thirty four oocytes remaining unfertilized after intracytoplasmic sperm injection were examined by fluorescence immunohistochemistry. Chromatin of injected sperm remained condensed without osH1 in 45% ; remained condensed with osH1 incorporated in 10% ; and had decondensed with osH1 incorporated in 45%. No oocyte showed decondensed sperm chromatin without osH1. Thus, osH1 incorporation into sperm chromatin precedes sperm chromatin decondensation, although whether incorporation of osH1 is needed for development after its disappearance is unclear. Immunohistochemical analysis in ovarian tissues with polyclonal anti-osH1 antibody revealed that the osH1 protein was localized in oocytes of all stages from primordial follicles through Graafian follicles. No reactivity of anti-osH1 antibody was noted in granulosa or theca cells. While the signal was more intense in the nucleus, the cytoplasm also was stained in oocytes examined at all stages. Full length cDNA of the os H1 was cloned, and the recombinant osH1 protein was expressed using wheat germ cell-free protein synthesis technology.

我们研究了osH1蛋白表达在人类受精中的生物学意义。针对代表osH1部分序列的合成肽产生的兔多克隆抗体用于卵母细胞和卵巢组织提取物的Western blotting，该抗体显示了一个36kDa的条带，这是osH1的计算分子质量。Western分析法通过双细胞胚胎检测萌发-囊泡期卵母细胞染色质中的蛋白。用荧光免疫组织化学方法检测了34个卵母细胞注射卵浆内精子后未受精的情况。注射后的精子中有45%的染色质没有osH1；加入10%的osH1后保持冷凝；并与osH1合并去凝，占45%。没有卵母细胞显示去致密的精子染色质，没有osH1。因此，进入精子染色质的osH1先于精子染色质去浓缩，尽管在其消失后发育是否需要加入osH1尚不清楚。卵巢组织免疫组化分析显示，osH1蛋白存在于从原始卵泡到Graafian卵泡的所有阶段的卵母细胞中。颗粒细胞和卵泡膜细胞无抗osh1抗体反应性。虽然信号在细胞核中更强烈，但在所有阶段检查的卵母细胞中细胞质也被染色。克隆了osH1基因的全长cDNA，利用小麦无生殖细胞蛋白合成技术表达重组osH1蛋白。

项目成果

Histone replacement of injected sperm head after ICSI.

ICSI 后注射精子头的组蛋白替换。

• 发表时间: 2006
• 作者: Mizusawa;Y;Kuji;N;Tanaka;Y;Yoshida;H;Hamatani;T;Yoshimura;Y;Kato;S;Komatsu;S
• 通讯作者: S

Fine resolution of human sperm nucleoproteins by two-dimensional electrophoresis.

• DOI: 10.1093/molehr/gah217
• 发表时间: 2005-09
• 期刊: Molecular human reproduction
• 影响因子: 4
• 作者: Tsuyoshi Yoshii;N. Kuji;S. Komatsu;K. Iwahashi;Yudai Tanaka;H. Yoshida;A. Wada;Y. Yoshimura

Post-transcriptional regulation may be involved in expression of oocyte-specific H1 and somatic H1s during human embryogenesis.

转录后调控可能参与人类胚胎发生过程中卵母细胞特异性 H1 和体细胞 H1 的表达。

• 发表时间: 2006
• 期刊: Hum Reprod 20(suppl 1)
• 作者: Yuri Mizusawa;Yudai Tanaka;Naoaki Kuji;Hiroyuki Yoshida;Shingo Kato;Yasunori Yoshimura
• 通讯作者: Yasunori Yoshimura

Related Articles, Links Protein kinase A activity and protein phosphorylation during the mouse sperm acrosomal reaction.

相关文章、链接 小鼠精子顶体反应过程中的蛋白激酶 A 活性和蛋白磷酸化。

• 发表时间: 2005
• 期刊: Arch Androl. 51(1)
• 作者: Kuji N;Tanaka Y;Komatsu S;Yoshimura Y
• 通讯作者: Yoshimura Y

Protein Kinase A Activity And Protein Phosphorylation During The Mouse Sperm Acrosomal Reaction

• DOI: 10.1080/014850190512743
• 发表时间: 2005-01
• 期刊: Archives of Andrology
• 作者: N. Kuji;Y. Tanaka;S. Komatsu;Y. Yoshimura