Title page
ACKNOWLEDGEMENT
Contents
Abstract 10
I. Introduction 11
II. Materials and methods 14
II. 1. Animals and reagents 14
II. 2. Preparation of mouse bone marrow hematopoietic cells 14
II. 3. Cell viability assay 14
II. 4. RT-PCR analysis 15
II. 5. Apoptosis assay of murine hematopoietic cells 16
II. 6. Antagonists assay 16
II. 7. Analysis of hematopoietic stem cells with flow cytometry 17
II. 8. Cell cycle analysis 17
III. Results 18
III. 1. ATP inhibits the proliferation of murine hematopoietic cells. 18
III. 2. P2X receptor family genes and P2X7 gene and protein are expressed on total hematopoietic cells and HSCs 20
III. 3. ATP induces intracellular caspase activation and apoptosis in hematopoietic cells. 24
III. 4. P2X7 receptor is mediated on extracellular ATP-induced apoptosis. 28
III. 5. ATP induces cell death in hematopoietic stem cells. 30
IV. Discussion 33
V. Conclusions 36
Reference 37
Summary in Korean 41
Figure. 1. ATP inhibits the proliferation of murine hematopoietic cells.To investigate influences of ATP on survival of murine hematopoietic cells, we performed survival assay by 19
Figure. 2. Expression of P2X receptor genes and P2X7 protein in hematopoietic cells including HSCs and progenitor cells. To investigate which P2XRs are expressed on the murine 22
Figure. 3. Extracellular ATP induces apoptosis of hematopoietic cells.To examine whether cell death was caused via apoptosis when stimulated by ATP, we carried out apoptosis 25
Figure. 4. ATP-induced apoptosis of hemtopoietic cells is involved in activaton of caspases. Cells were stimulated with different concentration of ATP for 3 days. After incubation, 27
Figure. 5. P2X7 receptor is mediated on extracellular ATP-induced apoptosis To confirm ATP-dependent cell death is specifically induced through P2X7 receptor, bone marrow cells 29
Figure. 6. ATP induces cell death in hematopoietic stem cells. To verify effects of extracellular ATP on HSCs, hematopoietic cells from mouse bone marrow were treated with the 31