EXPRESSION OF NEURONAL 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

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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 (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…………