Cell Membrane Transport, 1970 Principles and Techniques

It is not a particularly rewarding task to engage in writing a book on a subject which is undergoing a rapid and potentially revolutionary develop­ ment, but, on the other hand, the investigation of transport of substances into and out of cells has reached a stage of maturity or at least of self­ realization and this fact alone warrants a closer examination of the subject. No one will doubt at present that the movement-mostly by selective translocation-of substances, ranging from hydrogen ions to deoxyribo­ nucleic acids, across the cell-surrounding barriers represents one of the salient features of a living cell and that, if we are permitted to go so far, the cessation of the selective transport processes might be considered as the equivalent of cell death. Hardly anybody will question the premise that cell and tissue differentiation within the ontogenetic development of an organism is closely associated with properties of the outer cell face. Perhaps no serious scholar will attempt to refute the concept that mem­ branes with characteristic morphology and composition represent the ar­ chitectural framework for the whole cell. And probably no experienced biologist will raise objections to the belief that many physiological processes, like nervous impulse conduction and other electrical phenomena of cells and tissues or their volume changes, are associated with membrane-regulated shifts of ions and molecules.

Structural Aspects.- 1. Composition and Structure of Cell Membranes.- 1.1. Cell Envelopes and Membranes.- 1.2. Chemical Composition of the Plasma Membrane.- 1.3. Fine Structure of the Plasma Membrane.- Kinetic Aspects.- 2. Transport in Homogeneous Liquid Phase.- 2.1. Mass Flow and Diffusion.- 2.1.1. Introductory Definitions.- 2.1.2. The Laws of Diffusion.- 2.2. Migration and Electrodiffusion of Ions.- 3. Passive Membrane Transport of Nonelectrolytes.- 3.1. Permeation by Simple Diffusion.- 3.2. Mediated Diffusion and Related Phenomena.- 3.2.1. General considerations.- 3.2.2. Model I.- 3.2.3. Model II.- 3.2.4. Model III.- 3.2.5. Model IV.- 3.2.6. Inhibition of Mediated Diffusion.- 4. Permeation of Ions and Water.- 4.1. Permeation of Ions and Membrane Potentials.- 4.1.1. Introduction.- 4.1.2. Equilibrium of Ions across Membranes 92.- 4.1.3. Nonequilibrium Membrane Potentials and Ion Fluxes.- 4.2. Transport of Water.- 4.2.1. Ideal Semipermeable Membrane.- 4.2.2. Membrane Permeable to the Solute.- 5. Active and Coupled Uphill Transport.- 5.1. Active Transport.- 5.1.1. Criteria.- 5.1.2. Kinetics.- 5.2. Coupled Transport.- 6. Kinetics of Tracer Exchange.- 6.1. Introduction.- 6.2. Compartment in a Steady State, Communicating with One Other Compartment.- 6.2.1. Compartment and a Reservoir.- 6.2.2. Closed System of Two Compartments.- 6.3. Compartment in a Steady State, Communicating with Two Other Compartments.- 6.3.1. Compartment with Irreversible Outflow.- 6.3.2. Compartment with Reversible Outflow.- 6.4. Two Compartments and a Reservoir.- 6.4.1. Two Compartments in Parallel.- 6.4.2. Two Compartments in Series.- 6.5. Nonsteady-State Compartments.- 7. Special Types of Transport.- 7.1. Sorption Theories.- 7.2. Pinocytosis and Phagocytosis.- Molecular Aspects.- 8. Molecular Basis of Transport.- 8.1. Nature of Transporting Molecules.- 8.2. Movement of the Carrier.- 8.3. Uphill Transport Systems.- 8.4. Translocation by Metabolic Enzymes.- 8.5. Number of Carrier Sites per Cell.- 8.6. Isolation of Carrier Molecules.- 8.6.1. Double Labeling of Inducible Transport Proteins.- 8.6.2. Binding of Inhibitors to the Transport Protein.- 8.6.3. Search for Binding Affinity.- 8.7. Final Comment.- Methodological Aspects.- 9. Incubation and Separation Techniques.- 9.1. Incubation.- 9.1.1. Types of Incubators and Flasks.- 9.1.2. Gaseous Phases Used for Incubation.- 9.2. Separation of Cells and Tissues.- 9.2.1. HandHng of Macroscopic Objects.- 9.2.2. Handling of Microscopic Objects.- 10. Estimation of Solute Penetration.- 10.1. Analysis of the Incubation Medium.- 10.2. Water of Cells and Tissues.- 10.3. Intercellular Space.- 10.4. Estimation of Intracellular Composition.- 10.4.1. Preparation of Cells and Tissues for Analysis.- 10.4.2. Chemical Analysis.- 10.4.3. Radioactive Isotopes.- 10.5. Cytological Methods.- 10.5.1. Autoradiography.- 10.5.2. Other Techniques.- 10.6. Estimation of Solutes in Living Cells.- 10.6.1. Microelectrodes.- 10.6.2. Estimation of Intracellular pU by Acid Dye Distribution.- 10.7. Estimation of Membrane Transport by Nonspecific Methods.- 10.7.1. Densitometry.- 10.7.2. Indirect Cytolytic Method.- 11. Interpretation of Transport Data.- 11.1. Initial Rates.- 11.2. Inhibition of Transport.- 11.3. Activation of Transport.- 11.4. Some Less Common Cases.- 11.5. Analysis of Exponential Curves.- 11.6. Uptake of Metabolized Substrates.- 12. Bioelectrical Measurements.- 12.1. Introduction.- 12.2. Transepithelial Potential Differences and Short-Circuit Current Technique.- 12.3. Intracellular Potential Measurements Using Microelectrodes.- 12.4. Cation-Sensitive Glass Electrodes.- 13. Volume Flow Measurements.- 13.1. Volume Flow between Cell and Its Surroundings.- 13.1.1. Determination of Volume Flow from Cell Weight Changes.- 13.1.1. Estimation of Cell Volume.- 13.2. Transcellular Volume Flow and Flow across Cell Layers.- 13.2.1. Determination Based on the Change of Concentration of an Impermeant Substance.- 13.2.2. Determination Based on Weighing.- 13.2.3. Determination Based on Direct Volume Measurement.- 14. Use of Artificial Membranes.- 14.1. Introduction.- 14.2. Membrane Materials.- 14.3. Formation of Membranes.- 14.4. Design of Experimental Chambers.- 14.5. Electrical Measurements.- 14.6. Electrical Measurement Apparatus.- 14.7. Properties of Artificial Membranes.- 14.7.1. Electrical Properties.- 14.7.2. Permeability.- 15. Assay of Transport Proteins.- 15.1. Equilibrium Dialysis and Ultrafiltration.- 15.2. Ultracentrifugation.- 15.3. Column Chromatography.- 15.4. Conductometry.- 15.5. Other Methods.- Comparative Aspects.- 16. Bacteria.- 16.1. Introduction.- 16.2. Sugars.- 16.2.1. Escherichia coli.- 16.2.2. Other Bacteria.- 16.3. Amino Acids.- 16.3.1. Escherichia coli.- 16.3.2. Other Bacteria.- 16.4. Cations.- 16.5. Anions.- 17. Yeasts and Fungi.- 17.1. Introduction.- 17.2. Sugars.- 17.2.1. Monosaccharides.- 17.2.2. Oligosaccharides.- 17.3. Polyols.- 17.4. Amino Acids.- 17.5. Other Organic Compounds.- 17.6. Cations.- 17.7. Anions.- 18. Algae and Higher Plants.- 18.1. Introduction.- 18.2. Membrane Potentials.- 18.3. Ion Contents and Activities.- 18.4. Ion Fluxes.- 18.5. Active Transport.- 18.6. Sources of Energy for Transport.- 18.7. Effects of Light on Membrane Potential Differences.- 18.8. Ion Translocations across the Chloroplast Membrane.- 18.9. Electrical Properties.- 18.10. Transport of Water.- 18.11. Transport in Higher Plants.- 19. Erythrocytes.- 19.1. Introduction.- 19.2. Sugars.- 19.2.1. Monosaccharides.- 19.2.2. Disaccharides.- 19.3. Amino Acids.- 19.4. Other Organic Compounds.- 19.5. Cations.- 19.6. Anions.- 20. Muscle.- 20.1. Introduction.- 20.2. Morphology.- 20.3. Distribution of Ions and Membrane Potentials.- 20.4. Transport of Sugars and Amino Acids.- 21. Nerve.- 21.1. Introduction.- 21.2. Distribution and Fluxes of Ions.- 21.3. Passive Electrical Properties of Nerve Fibers.- 21.4. Action Potential and Its Propagation.- 21.5. Permeability to Nonelectrolytes.- 22. Epithelial Layers of Anurans.- 22.1. Actively Transported Ions.- 22.2. Origin of the Spontaneous Transepithelial Potential and Localization of the Potential Gradients.- 22.3. Intracellular Concentrations of Ions in Epithelial Layers. Active Transport at the Individual Membranes.- 22.4. Some Peculiarities Related to the Transport Properties of Anuran Epithelial Membranes.- 22.5. Ion Transport and Metabolism in Anuran Epithelial Membranes.- 22.6. Hormonal Control of the Transport Phenomena in the Anuran Epithelial Membranes.- 23. Intestine.- 23.1. Introduction.- 23.2. Sugar Absorption.- 23.2.1. Location of Disaccharidases.- 23.2.2. Monosaccharide Transport.- 23.3. Protein Absorption.- 23.3.1. Amino Acid Transport.- 23.3.2. Interaction between Amino Acid and Sugar Transport.- 23.4. Transport of Ions and Water.- 23.4.1. Monovalent Cations.- 23.4.2. Divalent Cations.- 23.4.3. Water.- 23.5. Interaction between the Transport of Nonelectrolytes and of Sodium.- 23.6. Absorption of Lipids.- 23.6.1. Pinocytosis.- 23.6.2. Diffusion.- 23.6.3. Chylomicrons.- 24. Kidney.- 24.1. Morphology.- 24.2. Experimental Techniques.- 24.3. Transport of Sodium Ions and of Water.- 24.4. Transport of Other Monovalent Ions.- 24.5. Transport of Calcium.- 24.6. Transport of Urea.- 24.7. Reabsorption of Sugars.- 24.8. Reabsorption of Amino Acids.- 25. Mitochondria.- 25.1. Introductory Remarks.- 25.2. Unspecific Permeable Space of Mitochondria.- 25.3. Transport of Cations.- 25.3.1. Monovalent Cations.- 25.3.2. Divalent Cations.- 25.4. Transport of Anions.- 25.4.1. Inorganic Anions.- 25.4.2. Organic Anions.- 25.4.3. Uncoupler Ions.- 25.5. Stoichiometry of Transport.- 25.6. Effect of Inhibitors.- 26. Tumor Cells.- 26.1. Sugars.- 26.2. Amino Acids.- 26.3. Ions.