Title Page

Contents

INTRODUCTION 8

MATERIALS AND METHODS 11

1. Ethical statement 11

2. Chemicals and Media 11

3. Preparation and Treatment of the spermatozoa with CK-636 12

4. Isolation of G-actin (Triton-soluble Cytoskeleton) and F-actin (Triton-insoluble Cytoskeleton) in spermatozoa 12

5. Computer assisted sperm analysis (CASA) 13

6. Combined Hoechst 33258/Chlortetracycline Fluorescence Assessment of Capacitation Status 14

7. Measurement of intracellular calcium ion concentration ([Ca2+]i)(이미지참조) 15

8. Western blot analysis of tyrosine phosphorylation of sperm proteins 15

9. In vitro fertilization (IVF) 16

10. Statistical analysis 17

RESULTS 18

1. Effect of CK-636 on Actin polymerization in spermatozoa 18

2. Effect of CK-636 on sperm motility and motion kinematics 20

3. Effect of CK-636 on capacitation status 22

4. Effect of CK-636 on level of [Ca2+]i.(이미지참조) 24

5. Effect of CK-636 on protein tyrosine phosphorylation 26

6. Effect of CK-636 on fertilization and embryonic development 28

DISCUSSION 31

REFERENCES 36

국문초록 49

ABSTRACT 51

Table 1. Effect of CK-636 on sperm motility and motion kinematics. 21

Figure 1. Effect of CK-636 on actin polymerization. 19

Figure 2. Effects of CK-636 on capacitation status. 23

Figure 3. Effects of CK-636 on [Ca2+]i.(이미지참조) 25

Figure 4. Effect of CK-636 on protein tyrosine phosphorylation. 27

Figure 5. Effect of CK-636 on fertilization and embryo development. 29

Figure 6. Effect of inhibition of Arp2/3 complex on embryo development. 30