Cell Physiology Source Book

Research Interests Research in his laboratory focuses on the molecular and cellular physiology of carrier protein molecules that actively transport chloride ions (Cl-) across the plasma membrane of neurons and epithelial cells. Specifically, they study some members of the cation-coupled-chloride contransporter gene/protein family SLC12A: the Na+, K+, 2 Cl- cotransporters (NKCC1 and NKCC2) and the K+-Cl- cotransporters (KCC1, 2, 3 and 4). These carrier proteins play key roles in: intracellular Cl- homeostasis in neurons, GABA- and glycine-mediated synaptic signaling, neuronal development, sensory transduction including nociception, transepithelial salt transport, cell water volume control, and extracellular K+ scavenging. Not surprisingly, altered function of these proteins underlies several pathologies and hence they have become significant targets for therapeutic interventions and translational research. To study the function of these proteins we use state-of-the-art live-cell imaging microscopy and fluorescent probes for measuring and manipulating intracellular ions and water in dissociated neurons and epithelial cells. Some of these optical methods have been developed in their lab, and are used in conjunction with molecular methods, knockout models, and several microanatomical techniques. Their current research involves two projects: Mechanisms regulating intracellular chloride in primary afferent neurons and their impact on GABA-mediated presynaptic inhibition and sensory transduction. This project aims at understanding the molecular mechanisms that determine intracellular Cl- concentration in primary afferent neurons, their regulation, and the role they play in presynaptic inhibition, acute somatic pain, neurogenic inflammation and proprioception. Roles of cation-coupled-chloride contransporters of choroid plexus epithelial cells in the regulation of cerebrospinal fluid ion composition. The choroid plexus epithelial cells (CPECs) form the blood-cerebrospinal fluid (CSF) barrier. CPECs secrete CSF and regulate its electrolyte composition. Regulation of CSF ion levels is fundamental for maintaining normal brain function. The overarching goal of this project is to understand how NKCC1, KCCs and aquaporins control the ion composition of the cerebrospinal fluid. Current emphasis is on the molecular and cellular mechanisms used by CPECs to regulate and maintain the CSF K+ concentration, a fundamental problem of broad physiological significance. CSF composition has a major impact on the fluid microenvironment of neurons and glial cells, and vice versa. Extracellular K+ homeostasis is critical for normal brain function; small changes in extracellular K+ profoundly affect neuronal excitability and osmotic water balance of glial cells and neurons. Dr. Eric Delpire teaches at the Vanderbilt University, Nashville, USA Edna Kaneshiro is a distinguished research professor in the department of biology at the University of Cincinnati where she has been for 43 years. Dr. Kaneshiro’s research is on the lipids of eukaryote protists, including free living, parasitic, and opportunistic pathogens. Although work on a number of different protozoa is being performed, Kaneshiro’s current focus is on the AIDS-associated opportunistic infection caused by Pneumocystis carinii. This organism causes a type of pneumonia that can lead to the death of immunocompromised individuals. Pneumocystis proliferates extracellularly in the lung alveolus where lipids constitute a major part of lung surfactant. Thus, lipids are thought to be important to the nutrition, physiology and metabolism of the organism. The biosynthesis of pathogen-specific lipids represents potential targets for drug development. She also has an interest in the cell biology and life history of this poorly understood opportunistic infectious agent. Dr. Kaneshiro is an elected fellow of the American Association for the Advancement of Science and the American Academy of Microbiology

SECTION I. Biophysical Chemistry, Cell Structure and Function 1. Water, Solutions, and Colligative Properties, Simple Diffusion, Permeability2. Lipids Artificial Membranes, Ionophores, Lipid Rafts, Liposomes3. Nucleic Acids Nucleus, Nucleolus, DNA, Chromosomes, Mitotic Spindle, Plasmids, Organellar DNA, Telomeres, DNA and RNA Editing, also see chapter 8)4. Proteins Cytoskeleton, Enzymes, Carriers, Protein Folding, Protein Crystallography5. Carbohydrates Glycocalyx, VSG, Mucins (e.g., T. cruzi), Receptors (Immune System, Lectins)6. Cell Membranes Bilayer Model, Lateral Mobility (Fluorescence Recovery After Photobleaching)7. Prokayotic Cells (eubacteria, archaea)8. Protists kDNA, Self-splicing, telomeres, apicoplasts, hydrogenosomes, mitosomes9. Organelles SECTION II. Membrane Potentials, Transmembrane Translocation 10. Membrane Potentials and Membrane Transport Gibbs-Donnan Equilibrium, Ion Channels in Non-Excitable Membranes, (Mutations, Disease, Toxin- & Drug-Binding)11. Carrier-Mediated Transport/Facilitated Diffusion Overview Ions (Na+, K+, Cl-), Sugars, Amino Acids, Nutrients, Drugs12. Na+/K+ ATPases13. Ca2+ Transport and Signaling (Surface Membrane, Cytosolic, Nuclear)14. Proton-Motive Force Ion Gradients, Chemosmotic Energy Transduction, Bacterial Flagella, Mitochondrial/Chloroplast Membrane Potential and Energy Production SECTION III. Membrane Excitability, Regulation of Ion Channels 15. Cable Properties, Length Constant, Electrogenesis of Membrane Excitability, Propagation of Action Potentials, Inhibition16. Voltage and Current Clamp (e.g., Oocytes, BLM), Patch-Clamp, Single Channel Techniques17. Voltage-Gated Channels18. Regulation of Mechanosensitive Channels, GTP, Cyclic nucleotides, G-proteins19. Nuclear Envelope SECTION IV. Integration, Synaptic Transmission, Sensory Transduction 20. Ligand-Gated Ion Channels21. Synaptic Transmission, Gap and Tight Junctions22. Excitation-Secretion Coupling, Trophic Effects of Nerves on Muscles? (see Endocrine?)23. Photoreceptors: Vision, Eye spots, Ocelli, Compound Eyes, Color Vision24. Mechanoreceptors: Sound, Balance, Stretch Receptors Pacinian Corpuscle, Hearing25. Pressure Receptors: Posture, Acceleration, Fish Lateral Line26. Chemoreceptors: Smell, Taste27. Electroreceptors, Magnetoreceptors (Magnetosomes), Thermoreceptors28. Brain, Glia SECTION V. Contractile Systems, Cell Locomotion, Muscle 28. Microtubule + Dynein + ATPase, Eukaryote Cilia39. Proton-Motive Force, Bacterial Flagella31. Spasmin/centrin + Ca2+, Rubber?32. Actin +Myosin + ATPase, Amoeboid Movements, Lymphocytes33. Skeletal Muscle, Excitation-Contraction Coupling, Sarcoplasmic Reticulum, Mechano-Chemistry, Insect click flight muscle?34. Smooth Muscle35. Cardiac Muscle SECTION VI. Integration, Endocrine, Exocrine Systems, Chemical Messengers, Hormones 36. Pituitary/Hypothalamus37. Adrenals38. Reproductive Physiology39. Respiration, Gas Transport, Respiratory Pigments, Acid-Base Balance40. Osmoregulation, Volume Regulation, Aquaporin, Ice/Freezing, Counter- Current Systems, Counter-Current Multiplier (Active Transport) Systems SECTION VII. SPECIALIZDED PROCESSES 41. Bioluminescence42. Plant Cell Physiology, Plastids, Photosynthesis, Stomata Opening43. Cell Migration, Quorum Sensing, Taxis, Kinesis, Tropism44. Biological Clocks/Chronobiology, Diapause, Hibernation, Brown Fat SECTION VIII. Experimental Methods/Approaches Cell Biology – Microscopy, GFP, Specific Stains, In Situ Hybridization, Light, Phase-Contrast; Fluorescence Microscopy, Fluorescence Resonance Energy Transfer (FRET); Multi-Photon, Electron Microscopy, Confocal, Imaging, FISH, Atomic Force, STORM/Single Molecule Imaging, Photoactivated Localization Microscopy (PALM); Total Internal Reflection Microscopy (TIRF), Fluorescence Recovery After PhotobleachingMolecular Biology –Nucleotide Sequencing, Whole Genome Analysis, Mutagenesis, Recombinant DNA, PCR, SDS-PAGE, Western Blots, Expression Systems, Antibody Preparation. Biochemistry - Enzyme Kinetics; Gas-Liquid Chromatography, Mass Spectrometary, Nuclear Magnetic Resonance Spectroscopy, Fourier Transforms, X-ray Scattering, Electron Spin Resonance SpectroscopyBiophysical – Optical Tweezers, Magnetic Resonance ImagingHigh Throughput – Bioinformatics, Genomics, Proteomics