Table of Contents
Acknowledgements .................................................................................................................. II
Abstract ................................................................................................................................... III
List of tables ........................................................................................................................... VI
List of figures ........................................................................................................................ VII
List of abbreviations ............................................................................................................ VIII
1.Introduction .................................................................................................................. 1
1.1. Muscle damage ................................................................................................... 2
1.2. Role of Macrophages ......................................................................................... 4
1.3. Macrophage Ablation ......................................................................................... 6
1.4. Macrophage depletion in mouse models ............................................................ 7
1.5. Mouse injury models ........................................................................................... 9
1.6. Rationale ............................................................................................................. 9
1.6.1Hypothesis ..................................................................................... 9
1.6.2Objectives...................................................................................... 9
2.Materials and Methods ............................................................................................ 11
2.1 Animal maintenance and treatments ............................................................... 12
2.1.1 Ethical Approval .......................................................................... 12
2.1.2 Treatment of mice with doxycycline ............................................ 12
2.1.3 Muscle injury and tissue harvest .................................................. 12
2.2 Processing tissue samples ............................................................................... 13
2.4. Histochemical and immunochemistry antibody stains ................................... 14
2.4.1 Haematoxylin and eosin staining ................................................. 14
2.4.2 F480 stain ..................................................................................... 14
2.4.3 CD68 antibody stain ..................................................................... 16
2.4.4 IgG stain ....................................................................................... 17
2.4.5 Laminin stain ................................................................................ 17
2.5 Statistical analysis ........................................................................................... 17
2.5.1 Morphometric analysis ................................................................. 18
2.5.2 Quantification of macrophages .................................................... 18
2.5.3 Calculation of fibre size diameter and central nucleation ............ 18
2.5.4 Percentage of tissue damage ........................................................ 18
3. Results ........................................................................................................................ 21