Synopsis
Section I Ultrastructure and Fragmentation of Neural Tissue.- 1 Bulk Separation of Neuronal Cell Bodies and Glial Cells in the Absence of Added Digestive Enzymes.- I. Introduction.- II. Bulk Isolation Procedures Requiring No Added Digestive Enzyme(s).- A. The Procedure Developed in the Authors' Laboratory.- B. The Procedure of Nagata et al. (1971).- C. The Procedure of Iqbal and Tellez-Nagel (1972).- D. The Procedure of Jones et al. (1971).- III. General Procedural Comments.- IV. Cell Yield and Biochemical Characterization.- V. Applications in Cellular Neurochemistry.- A. Centrifugal Fractionation of Isolated Nerve Cell Bodies.- B. Metabolic and Enzyme Studies.- C. Miscellaneous Applications.- Acknowledgments.- References.- 2 Isolation of Brain Mitochondria.- I. Introduction.- II. Isolation of Brain Mitochondrial Fractions by Differential Centrifugaron.- A. General Aspects.- B. Homogenization Techniques.- C. Isolation Media.- D. Method I: Isolation of Bovine Brain Mitochondria (Stahl et al., 1963).- E. Method II: Isolation of Rat Brain Mitochondria (Ozawa et al., 1966).- III. Fractionation of Crude Mitochondrial Fractions by Density Gradient Centrifugation.- A. General Aspects.- B. Method III: Isolation of Rat Brain Mitochondria (Tanaka and Abood, 1963).- C. Method IV: Isolation of Rat Brain Mitochondria in a Continuous Gradient by Isopycnic Centrifugation (van Kempen et al., 1965).- IV. Criteria of Purity of Mitochondrial Fractions.- A. General Aspects.- B. Electron Microscopy.- C. Enzymes as Markers for Mitochondrial Fraction from Brain.- References.- 3 Preparation of Brain Mitochondrial Membranes.- I. Introduction.- A. Definitions and Abbreviations.- B. Membrane Markers.- II. Methods for Fragmentation.- A. Hypotonic Procedures.- B. Hypotonic Procedures with Sonication.- C. Phospholipases.- D. Digitonin.- III. Enzymatic and Other Criteria.- Acknowledgments.- References.- 4 Isolation and Study of Synaptic Vesicles.- R. M. Marchbanks.- I. Introduction: Types of Vesicles.- II. General Comments on Isolation Methods.- A. Homogenization.- B. Separation of Synaptic Vesicles.- C. Monitoring the Synaptic Vesicle Fraction.- D. Determination of Bound Transmitter.- III. Procedures for the Isolation of Synaptic Vesicles.- A. Synaptic Vesicles from Cerebral Cortex and Related Tissues.- B. Cholinergic Vesicles from Electric Organ.- C. Adrenergic Vesicles from Heart, Vas Deferens, and Spleen.- IV. Studies on Isolated Synaptic Vesicles.- A. Biochemical Manipulations.- B. Morphological Examination.- References.- Section II Properties of Intact Neural Tissues.- 5 Culture of Neural Tissue and Cells.- I. Introduction.- II. Historical Background.- III. Methods of Primary Culture.- A. General.- B. Expiant Cultures of Neural Tissue.- C. Dissociated Cell Cultures of Neural Tissue.- D. Reaggregation Cultures of Neural Tissue.- IV. Methods for Clonal Cultures of Differentiated Tumors of the Nervous System.- A. Induction of Neural Tumors.- B. Primary Culture of Differentiated Neural Tumors.- C. Animal Passage of Primary Culture.- D. Subculture of Cultured Cells.- E. Cloning of Cultured Cells.- F. Culture Media.- G. Hybridization of Cultured Cells.- V. Studies with Primary Cultures.- A. Primary Expiants.- B. Dissociated Cells in Culture.- C. Reaggregation Cultures.- VI. Studies with Clonal Cells.- A. Glial Cells.- B. Neuroblastoma Cells.- VII. Conclusions.- Appendix: Preparation of Rat Tail Collagen.- Acknowledgments.- References.- 6 Measurement of Arteriovenous Differences Across Rat Brain.- I. Introduction.- II. General Considerations.- III. Procedures for Measurement of Arteriovenous Differences..- A. Anesthesia.- B. Cannulation of the Femoral Artery and Vein.- C. Cannulation of the Confluens Sinuum.- D. Blood Sampling and Metabolite Extraction.- E. Consideration of Blood Flow.- IV. Raising and Maintaining Blood Metabolite Concentrations.- A. General Considerations.- B. Approximation of the Priming Dose.- C. Maintenance of an Elevated Concentration.
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