Title Page
Contents
Abstract 6
I. Introduction 10
1. Obesity and chronic inflammation 10
2. Adipose Tissue Macrophage regulation during obesity 11
3. ATP Signaling in inflammation 13
4. Connexin-43 (Cx43) Gap junction 14
5. Purinergic Signaling 15
6. Inflammasome Activation by ATP via P2X receptors 16
7. Purpose of the current study 17
II. Materials and Methods 19
1. Animals 19
2. Western Blot Analysis 20
3. Quantitative PCR Analysis 21
4. Histology and immunohistochemistry 21
5. Stromovascular Cells (SVC) isolation and Flow cytometry 22
6. Cell Culture 23
7. Phagocytosis Assay 24
8. Immunocytochemistry 24
9. Statistical Analysis 25
III. Results 28
1. Macrophage Expression of Cx43 Was Upregulated in Gonadal White Adipose Tissue of HFD-Fed Mice 28
2. Macrophages Co-Cultured with Dying Adipocytes Upregulated Cx43 Expression 30
3. Cx43 Knockdown Reduced Inflammatory Response of Macrophages Co-cultured With Dying Adipocytes 32
4. Cx43 KO Protects Mice from HFD-Induced Metabolic Dysfunction, partly by Reducing the Inflammasome Signalling Pathway 36
IV. Discussion 42
V. Conclusion 44
VI. References 45
Table 1. Antibodies used for Western blot and Immunohistochemistry 26
Table 2. Primers used for qPCR 27
Figure 1. Cx43 is upregulated in gonadal white adipose tissue macrophages of mice by high-fat diet feeding 29
Figure 2. Macrophages co-cultured with dying adipocytes (dAC) upregulate Cx43 expression 31
Figure 3. Cx43 knockdown reduces inflammatory responses of macrophages co-cultured with dying adipocytes 34
Figure 4. Macrophage-specific Cx43 KO protected mice from HFD-induced inflammation in gonadal white adipose tissue 38
Figure 5. Macrophage-specific Cx43 KO protected mice from HFD induced metabolic dysfunction 40