Cell Biologist

Peter J. Hollenbeck and James R. Bamburg7

*Department of Biological Sciences

Purdue University

West Lafayette, Indiana 47907

^Department of Biochemistry and Molecular Biology Colorado State University Fort Collins, Colorado 80523

I. Introduction

II. Approaching Experimental Questions Using Neurons

A. Cell Behavior and Differentiation

B. Intracellular Events

III. Choosing a Neuronal Culture System

A. Basic Biological Properties

B. Developmental Properties

C. Source, Culture, and Handling Properties

D. Ease and Cost of Obtaining and Growing Neurons

IV. Conclusions References

Cell biologists of many stripes may find that their question of interest can be studied to advantage in neurons. However, they will also find that "neurons" include many and diverse cell types among which perhaps just one or a few may be ideal for a particular experiment. This chapter discusses the properties, relative complexity, and cost of primary neurons and neuronal cell types from different species and parts of the nervous system and compares their utility for different kinds of cell biological experiments.

METHODS IN CELL BIOLOGY, VOL. 71 Copyright 2003, Elsevier Science (USA). All rights reserved. 0091-679X/03 $35.00

I. Introduction

For the purpose of performing cell biological experiments, not all neurons are created equal. Both in vivo and in vitro, neurons from different parts of the nervous system are at least as varied in their properties as, for instance, epithelial cells. In addition, if we consider different species, including invertebrates, the range of cell biological properties becomes very wide. Choosing the correct neuron to study in culture requires the scrutiny of many cellular features, including cell size; neurite length, diameter, and type; growth cone size and rate of advance; biological responsiveness; and lifetime in culture. Important ancillary features that also figure in the choice of a system include the ease and cost of obtaining and maintaining neurons, and their accessibility to various experimental techniques, such as transfection, microinjection, or protein loading.

It is also worth noting here that not all parts of a single neuron are created equal. For example, axonal and dendritic growth cones of a polarized neuron show different motility behavior, can express unique receptors, and respond quite differently to environmental cues. This degree of spatial differentiation has been compared frequently—and only slightly naively—to the apical and basolateral domains of epithelial cells. Thus, cell biologists seeking to elucidate particular questions using neurons need not only choose the right neuron, but also the right end. In fact, neurons provide a polarized and compartmentalized system in which it is possible to separate and identify molecules that are unique to each domain (Job and Eberwine, 2001), something that cannot be accomplished for the cytoplasmic compartments of other polarized cells.

We attempt here to aid the experimentalist in choosing the right neuronal culture system. We first offer a general review of some major cell biological questions and the cultured neurons in which they have been studied effectively. Then, we survey a number of properties of different neuronal culture systems as a guide to weighing which systems suit particular experimental needs. Much of the information necessary to evaluate different culture systems is summarized in Tables I-IV. For a thorough treatment of the general principles of cell culture, we refer the reader to Volume 57 in this series.

II. Approaching Experimental Questions Using Neurons

Many cell biological problems have been pursued productively in one or more neuronal systems. Some examples follow.

A. Cell Behavior and Differentiation

1. Cell Crawling and Navigation

Many workers have studied the capacity of the neuronal growth cone to advance over a culture substratum and to respond to specific guidance cues such as

Table I

Names and Sources of Neurons and Cell Lines

Table I

Names and Sources of Neurons and Cell Lines

Was this article helpful?

0 0

Post a comment