Tissue Culture of the Nervous System

The impetus for compiling this book was the recent development of culture strains of neuroblastoma and glial cells and the immediate and enthusiastic way they have been taken up as model systems. After the first sudden rush of activity, it seems appropriate to pause, to assess progress, and to contemplate the future contributions that may be possible using these culture techniques. Long before the advent of established strains, cultures of nervous tissue had already contributed to neurobiology. Ross Harrison, in 1906, in a single experimental series, established tissue culture as a promising new technique in cell biology and settled the Golgi-Cajal controversy as to whether axonic processes originated as outgrowths from the cell body or were formed first in the intercellular spaces and were later connected to the cell body. Harrison observed process growth from nerve cells in cultures, thus settling the matter in favor of Cajal. Of great importance to neurobiology is the discovery by Rita Levi-Montalcini of nerve growth factor. Cultures of spinal ganglia played a major role in the discovery, isolation, and characterization of the factor (Levi-Montalcini et ai. , 1954). In my opinion, this discovery, although very well known, has not yet been adequately recognized for its germinal influence on neurobiology and embryology. Progress since the advent of clonal cultures has been more modest. I would like to cite two pieces of work which emphasize the technical ad­ vantages of these cultures.

Contenu

1 Long-Term Cultures of Embryonic and Mature Insect Nervous and Neuroendocrine Systems.- I. Introduction.- II. The Object of Choice: Periplaneta americana.- III. Material and Techniques.- IV. Specific Problems.- A. Cell Migration and Axonal Growth from Embryonic Brain and Ganglia.- B. Selective Outgrowth of Nerve Fibers and Interconnections In Vitro Between Thoracic Ganglia and Limb Bud Primordia.- C. In Vitro Studies on the Insect Neuroendocrine System.- V. Concluding Remarks and Perspectives.- VI. References.- 2 Differentiation of Aggregating Brain Cell Cultures.- I. Introduction.- II. Aggregation.- III. Migration.- IV. Biochemical Differentiation.- V. Morphological Differentiation.- VI. Prospectus.- VII. Acknowledgments.- VIII. References.- 3 Differentiation and Interaction of Clonal Cell Lines of Nerve and Muscle.- I. Introduction.- II. Origin of Cell Lines.- A. Neuroblastoma C1300.- B. Myoblast.- III. Morphological Differentiation.- A. Neuroblastoma.- B. Myoblast.- IV. Chemical Differentiation.- A. Neuroblastoma.- B. Myoblast.- V. Electrical Differentiation.- A. Neuroblastoma.- B. Myoblast.- VI. Differentiation and Cell Culture.- VII. Tropic Interaction Between Neuroblastoma and Myoblast Cell Lines.- VIII. Summary.- IX. References.- 4 Biochemical Characterization of a Clonal Line of Neuroblastoma.- I. Introduction.- II. Nucleic Acid and Protein Content.- III. RNA Biosynthesis.- A. Uridine Incorporation into RNA.- B. Sedimentation Pattern on Sucrose Gradient of Newly Synthesized RNA.- IV. Protein Biosynthesis.- A. Uptake and Incorporation of Leucine.- B. Analysis of Proteins by Polyacrylamide Gel Electrophoresis.- V. Glycoproteins of the Cell Surface.- VI. Discussion.- VII. References.- 5 Regulation of Neuronal Enzymes in Cell Culture.- I. Introduction.- II. Dissociated Primary Mouse Brain Cultures.- A. Monolayer Cultures.- B. Suspension-Rotation Cultures.- III. Neuroblastoma C1300 Ajax Mouse Cells in Culture.- IV. Neuroblast-Glioblast Biochemical Relationships.- A. Cyclic AMP.- B. Neurotransmitter Inactivation.- C. Glucose Transport.- V. Speculation.- A. In Vivo Neuronal Differentiation.- B. The Brain Code.- VI. References.- 6 Electrophysiological Studies of Normal and Neoplastic Cells in Tissue Culture.- I. Introduction.- II. The Mouse C1300 Neuroblastoma.- A. Electrical Excitability.- B. Chemosensitivity.- III. Hybrid Cell Lines.- IV. Other Continuous Cell Lines: Electrical Coupling.- V. Normal Dissociated Cultures.- A. Muscle Cultures.- B. Nervous System Cultures.- VI. References.- 7 Genetic Analysis of the Mammalian Nervous System Using Somatic Cell Culture Techniques.- I. General Methods for Genetic Analysis of the Nervous System.- II. Mitotic Inheritance of Differentiated Functions.- III. Hybridization Experiments.- A. General Considerations.- B. Neuroblastoma × L-Cell Matings.- C. Regulation of Adenosine-Cyclic-3?,5?-Monophosphate (c AMP).- D. Evidence for Pleiotropic Negative Control of Neuroectodermal Properties.- IV. Generation of Propagating Cell Lines from Normal Neurons and Glia.- V. Conclusions and Future Forecast.- VI. Acknowledgments.- VII. References.- 8 Nervous System-Specific Proteins in Cultured Neural Cells.- I. Introduction.- II. Experimental Neural Tumors.- III. S100 Protein Production by C6 Rat Glial Cells.- IV. Synthesis and Degradation of S100 Protein in C6 Cells.- A. Synthesis of S100 Protein by C6 Cells.- B. Degradation of S100 Protein in C6 Cells.- V. Production of 14-3-2 Protein by Human Neuroblastoma Cells in Culture.- VI. Conclusions.- VII. Acknowledgments.- VIII. References.- 9 Clonal Lines of Glial Cells.- I. Introduction.- II. Isolation of Clonal Lines.- A. General Approach.- B. Specific Approach.- III. Cellular Location of S100 Protein.- A. S100 Protein in Experimental Rat Tumors.- B. S100 Protein in Human Tumors.- C. S100 Protein in Clonal Cell Lines.- IV. Studies of Myelination in Vitro.- A. Demyelinating Disease.- B. Primary Tissue Culture Systems.- C. Clonal Line of Rat Peripheral Neurinoma Cells.- V. Prospectus.- VI. References.- 10 Induction of Enzymes by Glucocorticoids and Catecholamines in a Rat Glial Cell Line.- I. Introduction.- II. Materials and Methods.- III. Induction of Glycerol Phosphate Dehydrogenase by Cortisol.- IV. Effect of Catecholamines on Cyclic AMP and Lactic Dehydrogenase.- V. Differential Sensitivity of GPDH and LDH Inductions to Inhibitors of RNA Synthesis.- VI. Conclusions.- VII. References.- 11 The Metabolism of Glycosphingolipids and Glycosaminoglycans.- I. Introduction.- II. Glycosphingolipids.- A. Glycosphingolipids of the Nervous System.- B. Biosynthesis of Glycosphingolipids.- C. Catabolism of Glycosphingolipids.- D. Metabolism of Glycosphingolipids in Normal Nerve Cells.- E. Glycosphingolipid Metabolism in Cultured Cell Strains of Nervous System Origin.- III. Glycosaminoglycans.- A. Chemistry of Glycosaminoglycans.- B. Biosynthesis of Glycosaminoglycans.- C. Glycosaminoglycans in Nervous Tissue.- D. Production of Glycosaminoglycans in Cultured Cell Strains of Nervous System Origin.- IV. Collagen Synthesis.- V. Conclusions.- VI. References.