| 摘要 | 第3-6页 |
| Abstract | 第6-8页 |
| Chapter 1 Review | 第12-18页 |
| 1.1 Endochondral ossification | 第12-14页 |
| 1.1.1 The overview of endochondral ossification | 第12-13页 |
| 1.1.2 Regulation of endochondral ossification | 第13-14页 |
| 1.2 Sourecs and Toxicity of fluoride in the environment | 第14-16页 |
| 1.2.1 Sourecs of fluoride | 第14-15页 |
| 1.2.2 Fluoride toxicity in environment and its impact on skeletal system | 第15-16页 |
| 1.3 Bufo gargarizans is an excellent model to study endochondral ossification | 第16-18页 |
| Chapter 2 Hind limb of Bufo gargarizans as a model to study endochondral ossification during vertebrate metamorphosis | 第18-40页 |
| 2.1 Introduction | 第18-20页 |
| 2.2 Materials and methods | 第20-23页 |
| 2.2.1 Animals | 第20页 |
| 2.2.2 Determination of the skeletal development | 第20-21页 |
| 2.2.3 RNA extraction and cDNA library preparation | 第21页 |
| 2.2.4 Transcriptome sequencing, de novo assembly and functional annotation | 第21-22页 |
| 2.2.5 Differential gene expression analysis | 第22页 |
| 2.2.6 Quantitative real time RT-PCR validation | 第22页 |
| 2.2.7 Histological analysis | 第22-23页 |
| 2.2.8 In situ hybridization | 第23页 |
| 2.3 Results | 第23-35页 |
| 2.3.1 Development of hind limb | 第23-24页 |
| 2.3.2 RNA-sequencing, unigene assembly, and annotation | 第24-25页 |
| 2.3.3 Unigenes functional classification and metabolic pathway analysis | 第25-26页 |
| 2.3.4 Identification of differential gene expression among three different development stages | 第26-28页 |
| 2.3.5 Functional enrichment analysis of differentially expressed genes | 第28-30页 |
| 2.3.6 The expression profiles of genes related to the initiation of skeletal development | 第30-32页 |
| 2.3.7 Validation of DEGs by using qRT-PCR | 第32-33页 |
| 2.3.8 In situ hybridization of Ihh | 第33-34页 |
| 2.3.9 In situ hybridization of Sox9 | 第34-35页 |
| 2.4 Discussion | 第35-40页 |
| Chapter 3 The effects of excessive fluoride on endochondral ossification in hind limb of Bufo gargarizans | 第40-62页 |
| 3.1 Introduction | 第40-42页 |
| 3.2 Materials and methods | 第42-45页 |
| 3.2.1 Animals | 第42页 |
| 3.2.2 Water quality | 第42页 |
| 3.2.3 Determination of the skeletal development | 第42-43页 |
| 3.2.4 RNA extraction and cDNA library preparation | 第43页 |
| 3.2.5 Transcriptome sequencing, de novo assembly and functional annotation | 第43页 |
| 3.2.6 Differential gene expression analysis | 第43-44页 |
| 3.2.7 Quantitative real time RT-PCR validation | 第44页 |
| 3.2.8 Histological analysis | 第44页 |
| 3.2.9 In situ hybridization | 第44-45页 |
| 3.3 Results | 第45-56页 |
| 3.3.1 Water quality | 第45页 |
| 3.3.2 Development of hind limb | 第45-47页 |
| 3.3.3 RNA-sequencing, unigene assembly, and annotation | 第47-48页 |
| 3.3.4 Sequence annotation and functional classification | 第48-49页 |
| 3.3.5 Identification and functional classification of differentially expressed unigenes | 第49页 |
| 3.3.6 Functional enrichment analysis of differentially expressed genes | 第49-51页 |
| 3.3.7 The expression level of genes related to the initiation of skeletal development | 第51-53页 |
| 3.3.8 The effect of fluoride on THs-related genes expression | 第53-54页 |
| 3.3.9 The effect of fluoride on Ihh and Sox9 expression | 第54页 |
| 3.3.10 The effect of fluoride on BMP6, FGF3 and Wnt10b expression | 第54-55页 |
| 3.3.11 In situ hybridization of Ihh | 第55页 |
| 3.3.12 In situ hybridization of Sox9 | 第55-56页 |
| 3.4 Discussion | 第56-62页 |
| Chapter 4 Conclusion | 第62-64页 |
| References | 第64-74页 |
| 致谢 | 第74-76页 |
| 攻读硕士学位期间研究成果 | 第76页 |
