THE ROLE OF PARATHYROID HORMONE-RELATED PROTEIN IN REGULATING NEONATAL LUNG DEVELOPMENT by

vii CHAPTER I: INTRODUCTION Mammal lung development 1 Lung myofibroblasts and bronchopulmonary dysplasia disease 4 Parathyroid hormone-related protein 5 PTHrP signal transduction pathways 6 CHAPTER II: MATERIALS AND METHODS Mouse strains and breeding 11 Histology and morphometric analysis 13 Tissue total RNA isolation and reverse transcription 14 Real-time PCR analysis 14 Immunohistochemistry 15 Cell proliferation 15 Tissue lysis and subcellular fractionation 16 Western blot 16 Data presentation and statistical analysis 17 CHAPTER III: RESULTS AND DISCUSSION Conditional abrogation of PTHrP in mouse lung mesenchyme 18 The growth retardation and gross abnormalities in mesenchymal-specific 20 PTHrP knock-out mice Conditional abrogation of PTHrP in lung epithelia does not affect lung 23 development Failure of alveogenesis and severe bronchopulmonary dysplasia-like 26 changes in mesenchymal-specific PTHrP knock-out mice Significant reduction of cell proliferation with dysregulation of cell 31 differentiation in mesenchymal-specific PTHrP knock-out mouse lungs Loss of lung myofibroblasts in mesenchymal-specific PTHrP knock-out 33 mouse lungs iv Excessive and disordered elastin accumulated in mesenchymal-specific 38 PTHrP knock-out mouse lungs Intracrine PTHrP regulates cell proliferation 40 Discussion 41 Conclusion 46