mice 43 3.10 Adipocyte size in the obese Hp

I

INDEX

ABSTRACT III

1. INTRODUCTION 1

1.1 Obesity 2

1.2 Physiological role of white and brown adipose tissue 3

1.3 The evolution of the white adipose tissue concept 5

1.4 How obesity regulates the expression of key adipokines 8

1.5 Obesity and inflammation 9

1.6 Haptoglobin in the adipose tissue 11

1.7 What is Haptoglobin 12

1.7.1 Expression and regulation 12

1.7.2 Genetics and structure 12

1.8 Currently known functions of Haptoglobin 13

1.9 Aim 15

2. MATERIALS AND METHODS 16

2.1 Experimental animals 17

2.2 Histology, immunohistochemistry and morphometric analysis of tissues 18

2.3 Ex-vivo cultures and primary adipocytes 18

2.4Culture, differentiation and treatment of 3T3-L1 pre-adipocytes 19

2.5 MEFs isolation, culture, proliferation rate and differentiation process 20

2.6 Oil Red O staining 21

2.7 Assessment of triglyceride content and liver triglyceride content 21

2.8 Isolation of total RNA and Real-Time PCR 22

2.9 Isolation of gDNA 23

II

2.10 Glucose tolerance, insulin sensitivity and ELISAs 23

2.11 Protein isolation 23

2.12 In vivo insulin signaling 24

2.13Microarray analysis 24

2.14 Statistical analysis 25

3. RESULTS 26

3.1 Hp release by mouse WAT 27

3.2 HFD results in increased Hp production in mice 28

3.3 Physical parameters and plasma biochemistry of Hp^−/− mice 30

3.4 Hp deficiency results in ameliorated glucose homeostasis in obesity 33

3.5 Insulin action in liver, WAT and skeletal muscle of HFD Hp^−/− mice 34

3.6Liver in lean and obese Hp^−/− mice 36

3.7Adiponectin is increased in HFD Hp^−/− mice 40

3.8Effect of Hp on adiponectin expression in vitro 41

3.9WAT macrophage content in Hp^−/− mice 43

3.10 Adipocyte size in the obese Hp^−/− mouse 45

3.11 Adipocyte size in the lean Hp^−/− mouse: comparison with the obese 47

3.12 Hp deficiency and WAT gene expression 50

3.13 Hp and adipogenesis 54

4. DISCUSSION 58

5. BIBLIOGRAPHY 66

ACKNOWLEDGMENTS