EXPRESSION OF NEURONAL PROTEINS IN A
DIFFERENTIATING HUMAN NEUROBLASTOMA CELL LINE
(IMR32): INSIGHTS INTO NEURONAL DEVELOPMENT AND
DISEASE
by
YA CHEN
Submitted in partial fulfillment of the requirements
For the degree of Doctor of Philosophy
Dissertation Advisers: Dr. Maureen W. McEnery
Dr. Rolfe G. Petschek
Department of Physics
CASE WESTERN RESERVE UNIVERSITY
January, 2006
CASE WESTERN RESERVE UNIVERSITY
SCHOOL OF GRADUATE STUDIES
We hereby approve the dissertation of
______
candidate for the Ph.D. degree *.
(signed)______(chair of the committee)
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(date) ______
*We also certify that written approval has been obtained for any proprietary material contained therein. Table of Contents
Table of Contents………………………………………………………………….………………………… i
Dedication………………………………………………………………………………………………………… iv
List of Figures………………………………………………………………………………….……………….. v
Acknowledgement……………………………………………………………………….…….…………… ix
List of Abbreviations…………………………………………………………………………………….…… x
Abstract………………………………………………………………………………………………………….…… xiii
Chapter 1
Introduction
1.1 Neuronal differentiation ………………………………………………….………..…..………..… 2
1.1.1 Human neuroblastoma cell line (IMR32 cells) …………..………………..…….………..… 3
1.1.2 Differentiation of IMR32 cells…………………..……………………………………………..………… 3
1.2 Neuronal proteins …………………………………..…..……………………………………………..… 5
1.2.1 NCAM ……………..…………………………………………...…………………….………………….…… 6
1.2.2 MeCP2…………………………………………………………………………………………………………. 8
1.2.3 Voltage dependent calcium channel (VDCC)………………………………………..…….…… 9
1.2.3.1 High voltage dependent calcium channel ……………..…………………………………….. 10
1.2.3.2 Low voltage dependent calcium channel ………..………………………………………..….. 16
1.2.3.3 VDCC and Disease …………..…………………………………………………………..……….. 21
1.3 Proto-oncogene (N-myc) ……………………………………………………….…………..……….. 22
i 1.4 Expression of markers of neuronal development (MeCP2, NCAM, and VDCC subunits) in differentiated IMR32 cells……………….………………………………………..…….. 23
Chapter 2
Expression of neuronal proteins parallels neurotypic differentiation of human neuroblastoma cells (IMR32) …………………..…………………………….…… 30
2.1 Abstract…………………..………………………………………………………………….………………… 31
2.2 Introduction…………………..………………………………………………………………………….…… 33
2.3 Methods and Materials………………………………...…..…………………………………………… 36
2.4 Results…………