Identification of the potential hair cell regeneration ability of apical Lgr5+ cells in the neonatal murine cochlea

摘要

小鼠耳蜗的支持细胞是毛细胞损伤后再生的可靠来源。Lgr5+阳性的支持细胞是耳蜗中富含毛细胞的祖细胞的群体，它对揭示祖细胞增殖和分化的调节机制尤为重要。大量研究显示，顶圈和底圈的支持细胞增殖和分化为毛细胞的能力存在明显差异，然而，顶圈和底圈支持细胞转录组的差异表达并未被探讨过。因此，我们通过流式细胞仪分选出Lgr5+细胞，发现顶圈的Lgr5+细胞与底圈Lgr5+细胞相比分化出的毛细胞明显增多，具有更高增殖和分化潜能。同时，顶圈Lgr5+细胞能形成更多的神经球和更好的传代。然后，我们用微阵列分析顶圈和底圈Lgr5+细胞转录组中富集的和差异表达的基因，以及细胞周期基因和参与调节Lgr5+祖细胞增殖分化的转录因子。最后，分析差异表达的基因的作用，绘制耳蜗毛细胞再生的基因网络和蛋白相互作用网络。我们的数据库展示了Lgr5+祖细胞中参与调节细胞增殖和分化为毛细胞的可能基因，这些基因可能对未来毛细胞再生提供新的治疗靶点。

目录

声明

Abstract

摘要

Chapter Ⅰ Introduction

1．1．Overview

1．2．Hair cell regeneration potential of resident SCs in the inner ear

1．3 Lgr5 marked SCs in the mammalian inner ear

1．4 Factors affecting auditory HC regeneration after damage in mammals

1．5 Epigenetic regulation in HC regeneration

1．6 The role of Wnt signaling pathway in auditory HC development and regeneration

1．7 Implication of canonical Wnt／β-catenin signaling in auditory HC regeneration

1．8 Microarray Technology

1．9 Aims of the study

Chapter Ⅱ In vivo lineage tracing of apical and basal Lgr5+ cells

2．1 Introduction

2．2 Materials and Methods

2．3 Resulb and discussion

Chapter Ⅲ HC regeneration potential of apical and basal LgrS+cells in vitro

3．1 Introduction

3．2 Materials and Methods

3．3 Results and discussion

Chapter Ⅳ Neurosphere formation and differentiation of apical and basal Lgr5+cells in vitro

4．1 Introduction

4．2 Materials and Methods

4．3 Results and discussion

Chapter Ⅴ Transcriptome expression profiles of apical and basal Lgr5+ cells

5．1 Introduction

5．2 Materials and Methods

5．3 Results and discussion

Chapter Ⅵ Project summary and Future work plan

6．1 Summary and conclusions

6．2 Future directions

Appendix

Acknowledgement

References

Resume