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Title page
ABSTRACT
국문요약
List of abbreviations
Contents
INTRODUCTION 19
MATERIALS AND METHODS 26
1. Chemicals 26
2. Dissociation of rat cerebellar Purkinje neurons 27
3. Ca + imaging 28
4. Antisense oligodeoxynucleotides (AS-oligos) 30
5. Electrophysiological studies 33
6. Organotypic slice cultures of cerebellum 34
7. Immunofluorescence 35
8. Confocal microscopy 36
9. Statistics 37
RESULTS 38
1. Functional interaction between NCX and mGluR1 in Purkinje neurons 38
1.1. DHPG-induced [Ca +]i changes in isolated Purkinje neurons 38
1.2. Effects of NCX inhibitors on DHPG-induced increases of [Ca +]i in Purkinje neurons 46
1.3. Effects of suppression of NCX expression on DHPG-induced [Ca +]i responses in Purkinje neurons 51
2. Functional interaction between NCX and AMPA receptor in Purkinje neurons 61
2.1. Effects of NCX inhibition on AMPA-induced [Ca +]i changes in Purkinje neurons 61
2.2. Effects of NCX inhibitors on AMPA-induced currents in Purkinje neurons 71
3. The role of NCX on neuronal development of Purkinje neurons in slice cultures 76
3.1. Cerebellar slice culture 76
3.2. Effects of NCX activity on dendritic development of Purkinje neurons in cerebellar slice cultures 81
DISCUSSION 87
REFERENCES 96
감사의 글 108
Table 1. NCX antisense oligodeoxynucleotides (AS-oligos). 32
Table 2. AMPA-induced Ratio340/380 changes in Purkinje neurons. 69
Table 3. Effects of NCX AS-oligos on AMPA-induced Ratio340/380 changes in Purkinje neurons. 70
Fig. 1. DHPG-induced transient [Ca +]i changes in dissociated Purkinje neurons. 41
Fig. 2. Effect of CPCCOEt on DHPG-induced transient [Ca +]i changes in Purkinje neurons. 43
Fig. 3. Effects of NBQX on DHPG-induced [Ca +]i changes in Purkinje neurons. 45
Fig. 4. Effects of bepridil on DHPG-induced [Ca +]i changes in Purkinje neurons. 48
Fig. 5. Effects of KB-R7943 on DHPG-induced [Ca +]i changes in Purkinje neurons. 50
Fig. 6. Effects of lipofectamin on DHPG-induced [Ca +]i changes in Purkinje neurons. 54
Fig. 7. Effects of NCX S-oligos and AS-oligos on DHPG-induced [Ca +]i changes in Purkinje neurons. 56
Fig. 8. Effects of SKF 96365 on DHPG-induced [Ca +]i changes in Purkinje neurons. 58
Fig. 9. Effects of 0 Ca + solution on DHPG-induced [Ca +]i changes in Purkinje neurons. 60
Fig. 10. Effects of CPCCOEt on AMPA-induced transient [Ca +]i changes in Purkinje neurons. 64
Fig. 11. Effects of NCX inhibitors on AMPA-induced transient [Ca +]i chnges in Purkinje neurons. 66
Fig. 12. Effects of NCX AS-oligos and S-oligos on AMPA-induced transient [Ca +]i change in Purkinje neurons. 67
Fig. 13. Transmembrane current via NCX in Purkinje neurons. 73
Fig. 14. AMPA-activated currents in Purkinje neurons. 74
Fig. 15. Effects of NCX inhibitors on AMPA-activated current responses in Purkinje neurons. 75
Fig. 16. Images of Purkinje neurons in cerebellar slice cultures after DIV 1. 77
Fig. 17. Images of Purkinje neurons in cerebellar slice cultures after DIV 6. 79
Fig. 18. Images of Purkinje neurons in cerebellar slice cultures after DIV 12. 80
Fig. 19. Cerebellar slice cultures at DIV8. 83
Fig. 20. Effects of PKC activation on dendritic development of Purkinje neuron in cerebellar slice cultures. 85
Fig. 21. Effects of NCX inhibition on dendritic development of Purkinje neuron in cerebellar slice cultures. 86
Fig. 22. Functional interaction between NCX and glutamate receptors (mGluR1 and AMPAR) modulating [Ca +]i in Purkinje neurons. 95
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