Gangliosides 자극에 의한 신경소교세포의 형태학적 변화에서의 tubulin의 역할에 대한 연구
The role of tubulin in ganglioside-induced ramifications in cultured rat brain microglia
- 주제(키워드) tubulin+actin+gangliosides+microglia+ramification
- 발행기관 아주대학교
- 지도교수 주 일 로
- 발행년도 2006
- 학위수여년월 2006. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의학과
- 본문언어 영어
초록/요약
In addition to functional changes, activated microglia undergo morphological changes from a resting, ramified morphology to an activated, amoeboid shape. Gangliosides, despite their role as microglial activators, induce microglial ramifications. To identify the cytoskeletal components involved in microglial ramification, we stimulated cells with gangliosides in the absence or presence of cytoskeletal inhibitors and examined the resultant morphological changes. While LPS-induced ameboid shapes were blocked by all inhibitors, actin inhitibors and microtubule inhibitor, Ganglioside-induced ramifications were blocked by either paclitaxel or nocodazole, inhibitors of microtubule, but not by cytochalasin B or latrunculin B, which are inhibitors of actin polymerization. In contrast to these morphological changes, iNOS expression was suppressed by both inhibitors of actin filament and inhibitors of microtubule. These results indicate that ganglioside-induced ramification is more dependent on tubulin than actin and that both of these cytoskeletal components are involved in iNOS expression.
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ABSTRACT i
TABLE OF CONTENTS ii
LIST OF FIGURES iv
ABBREVIATION v
I. INTRODUCTION 1
A. Brain inflammation 1
B. Gangliosides 2
C. Cytoskeleton 3
D. Aims of study 4
II. MATERIALS AND METHODS 6
A. Reagents 6
B. Cell culture 6
C. Immunostaining and confocal microscopy 7
D. Phase contrast microscopy 7
E. Western Blot Analysis 7
F. Enzyme-linked immunosorbent assay (ELISA) 8
G. Data analysis 8
III. RESULTS 9
A. Gangliosides induce a unique microglial morphology with branching that is different from those induced by LPS or IFN-γ. 9
B. Gmix-induced morphology is not blocked by actin depolymerization. 13
C. Gmix-induced morphology is blocked by microtubule disruption. 19
D. LPS-induced morphology is blocked by inhibition actin filaments and microtubule. 24
E. Inhibition of actin filament or microtubules are suppressed cellular functions in Gmix-stimulated microglia. 29
IV. DISCUSSION 32
V. CONCLUSION 35
REFERENCES 36
국문요약 41
