上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

HDAC1基因决定涡虫成体干细胞的维持

张振超，曾 安，韩晓帅，李永芹，荆 清   

  1. 上海交通大学医学院/中科院上海生命科学研究院健康科学研究所核酸分子医学研究组， 上海 200025
  • 出版日期:2013-09-28 发布日期:2013-09-29
  • 作者简介:张振超(1987—)，男，硕士; 电子信箱: zhenchaozhang@163.com。
  • 基金资助:

    国家自然科学基金重点项目（81130005）；国家重点基础研究发展计划（2010CB945600)

HDAC1 determines adult stem cell maintenance in planarian

ZHANG Zhen-chao, ZENG An, HAN Xiao-shuai, LI Yong-qin, JING Qing   

  1. Laboratory of Nucleic Acid & Molecular Medicine, Institute of Health Sciences, Shanghai Jiaotong University School of Medicine, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China
  • Online:2013-09-28 Published:2013-09-29
  • Supported by:

    National Natural Science Foundation of China，81130005； National Basic Research Program of China, 2010CB945600

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摘要：

目的 分析涡虫HDAC1基因的表达谱，并研究HDAC1基因在涡虫稳态维持和再生中的功能及机制。方法 利用cDNA末端快速扩增技术获得涡虫HDAC1基因的全长序列，利用全组织原位杂交技术和Western blotting技术分别检测HDAC1转录本和蛋白在涡虫中的表达谱。通过RNA干扰技术干扰涡虫HDAC1基因的表达水平，并观察涡虫在组织稳态维持和再生过程中与对照组的区别，进一步利用免疫荧光实验检测对涡虫成体干细胞的影响。结果 成功克隆了涡虫HDAC1基因的全长序列；全组织原位杂交和Western blotting结果显示HDAC1基因在涡虫成体干细胞中高表达；HDAC1基因干扰后涡虫正常稳态维持不能进行，再生过程受到明显抑制；干细胞分子标记的免疫荧光实验揭示涡虫成体干细胞数量严重降低。结论 HDAC1基因在涡虫成体干细胞维持和促进涡虫再生中起着重要作用。

关键词: 涡虫, 再生, HDAC1, 成体干细胞

Abstract:

Objective To analyze the expression of HDAC1 in planarian, and explore the function and mechanism of HDAC1 in maintaining tissue homeostasis and planarian regeneration. Methods The full-length sequence of HDAC1 was acquired by rapid amplification of cDNA ends. The expression pattern of HDAC1 was determined by whole-mount in situ hybridization and Western blotting. The phenotype of planarian in tissue homeostasis and regeneration was determined after depleting HDAC1 by RNA interference. The expression of adult stem cell marker was assessed by immunofluorescence. Results The full-length sequence of HDAC1 was successfully cloned. Whole-mount in situ hybridization and Western blotting revealed that both HDAC1 transcripts and protein were highly enriched in planarian adult stem cells. Upon HDAC1 knockdown, planarian failed to maintain tissue homeostasis and regenerate missing body parts. Immunofluorescence with a stem cell marker showed that the number of adult stem cell also decreased significantly after HDAC1 knockdown. Conclusion HDAC1 gene plays important roles in maintaining planarian adult stem cells and promoting tissue regeneration.

Key words: planarian, regeneration, HDAC1, adult stem cells