1. Topic 3 The Neuronal Membrane at Rest Lange Biology 463 - Neurobiology

2. Introduction Resting Potential – the chemical makeup of the neuron when it is at “rest”…. meaning there is no message present Action Potential – the chemical makeup of the neuron when it is “active”…. meaning there is a message present Please note that the image above displays neurons in a way that can perpetuate a misconception: The Action Potential does NOT occur throughout the entire neuron instantly, but instead… the Action Potential will exist in a small portion of the neuron and will travel along the length of the neuron.

3. The Cast of Chemicals Involved the Generation of an Action Potential Cytosolic and Extracellular Fluids –water is a polar solvent –Various cations (+) and anions (-)

4. The Phospholipid Membrane –Hydrophilic (Polar) Head Materials that dissolve in water will pass through –Hydrophobic (Nonpolar) Tails Materials that dissolve in water will NOT pass through Materials that dissolve in non-polar solvents (such as oils) will pass through

5. –The phospholipids will aggregate in a water media to create what is known as the Phospholipid Bilayer

6. With the phospholipid molecule having the polar head region and the nonpolar tail region, when the phospholipid bilayer (PLB) is formed, the property that is seen is a PLB that serves as a selectively permeable membrane… a crucial feature for message transport both in the “electrical message” and the chemical message aspect of the neuron.

7. Proteins –Molecules Enzymes Cytoskeletal elements Receptors Specialized transmembrane proteins –Control resting and action potentials It is estimated that there can be hundreds of billions of different types of proteins that can be made, and our best estimate is that there are roughly 100,000 different proteins produced and used in the human body.

8. Protein structure is enormously diverse: The 20 different amino acids consisting of an alpha carbon, a carboxyl group, hydrogen, and some sort of R (residue) group Diversity is due to the variation in the R group

9. In the neuron, proteins are synthesized by ribosomes in two primary areas: –Neuronal cell body (soma) –Axon terminals In both places, the ribosomes assemble amino acids into polypeptide chains (connected by polypeptides)

10. Levels of protein structure. Secondary structure: The primary chain forms spirals (  -helices) and sheets (  -sheets). Tertiary structure: Superimposed on secondary structure.  -Helices and/or  -sheets are folded up to form a compact globular molecule held together by intramolecular bonds. Quaternary structure: Two or more polypeptide chains, each with its own tertiary structure, combine to form a functional protein. Tertiary structure of prealbumin (transthyretin), a protein that transports the thyroid hormone thyroxine in serum and cerebro- spinal fluid. Quaternary structure of a functional prealbumin molecule. Two identical prealbumin subunits join head to tail to form the dimer. Amino acid  -Helix: The primary chain is coiled to form a spiral structure, which is stabilized by hydrogen bonds.  -Sheet: The primary chain “zig-zags” back and forth forming a “pleated” sheet. Adjacent strands are held together by hydrogen bonds. (a) Primary structure: The sequence of amino acids forms the polypeptide chain. (b) (c) (d)

11. (a) Primary structure: The sequence of amino acids forms the polypeptide chain. Amino acid

12.  -Helix: The primary chain is coiled to form a spiral structure, which is stabilized by hydrogen bonds.  -Sheet: The primary chain “zig-zags” back and forth forming a “pleated” sheet. Adjacent strands are held together by hydrogen bonds. (b) Secondary structure: The primary chain forms spirals (  -helices) and sheets (  -sheets).

13. Tertiary structure of prealbumin (transthyretin), a protein that transports the thyroid hormone thyroxine in serum and cerebro- spinal fluid. (c) Tertiary structure: Superimposed on secondary structure.  -Helices and/or  -sheets are folded up to form a compact globular molecule held together by intramolecular bonds.

14. Levels of protein structure. Quaternary structure of a functional prealbumin molecule. Two identical prealbumin subunits join head to tail to form the dimer. (d) Quaternary structure: Two or more polypeptide chains, each with its own tertiary structure, combine to form a functional protein.

15. An example of the progression in complexity of structure in proteins with the final quaternary structure being that of hemoglobin.

17. Proteins develop in a variety of forms in neurons to form –Ion channels using channel proteins –Ion pumps using other proteins

18. The Movement of Ions Simple diffusion is an option for small ions and other small molecules

19. Electrical current influences ion movement Electrical conductance (g) and resistance (R); R = 1/g Electrical potential (voltage) Electricity and Circuits

20. Chemicals Involved in the Conduction of Electricity Electricity –Electrical current can flow across a membrane –Ohm’s law I = gV (current = conductance x electrical potential)

21. The Ionic Basis of The Resting Membrane Potential Membrane potential: Voltage across the neuronal membrane

22. Equilibrium Potential (E ion ) –No net movement of ions when separated by a phospholipid membrane –Equilibrium reached when K + channels inserted into the phospholipid bilayer –Electrical potential difference that exactly balances ionic concentration gradient

23. Equilibrium Potential

24. Voltmeter Microelectrode inside cell Plasma membrane Ground electrode outside cell Neuron Axon

25. Depolarizing stimulus Membrane potential (voltage, mV) Time (ms) 0 –100 –70 0 –50 +50 1234567 Hyperpolarizing stimulus Membrane potential (voltage, mV) Time (ms) 01234567 –100 –70 0 +50 Inside positive Inside negative (a)(b) Resting potential Depolarization Resting potential Hyper- polarization

26. (b) Depolarized regionStimulus Plasma membrane Depolarization Spread of depolarization (a)

27. Distance (a few mm) –70 Resting potential Active area (site of initial depolarization) Membrane potential (mV)

28. Copyright © 2010 Pearson Education, Inc. 01234 –70 –55 0 +30 Membrane potential (mV) Time (ms) Relative membrane permeability Na + K+K+ K+K+ Outside cell Inside cell Outside cell Inside cell Depolarizing phase: Na + channels open Repolarizing phase: Na + channels inactivating, K + channels open Action potential P Na PKPK Threshold Na + K+K+ K+K+ Outside cell Inside cell Outside cell Inside cell Inactivation gate Activation gates Potassium channel Sodium channel Resting state: All gated Na + and K + channels closed (Na + activation gates closed; inactivation gates open) Hyperpolarization: K + channels remain open; Na + channels resetting 2 2 3 4 4 1 1 1 3

29. –70 +30 (a) Time = 0 ms(b) Time = 2 ms(c) Time = 4 ms Voltage at 2 ms Voltage at 4 ms Voltage at 0 ms Resting potential Peak of action potential Hyperpolarization Membrane potential (mV))

30. Time (ms) Voltage Membrane potential (mV) –70 0 +30 Threshold Action potentials Stimulus amplitude

31. Stimulus Membrane potential (mV) Time (ms) –70 0 +30 012345 Absolute refractory period Relative refractory period Depolarization (Na + enters) Repolarization (K + leaves) After-hyperpolarization

32. Equilibrium Potentials (Cont’d) –The Nernst Equation Calculates the exact value of the equilibrium potential for each ion in mV Takes into consideration: –Charge of the ion –Temperature –Ratio of the external and internal ion concentrations

33. The Distribution of Ions Across The Membrane

34. The sodium-potassium pump –Enzyme - breaks down ATP when Na present –Calcium pump: Actively transports Ca 2+ out of cytosol

35. Na + K+K+ K+K+ K+K+ K+K+ Cell interior Na + 15 mM K + 150 mM Cl – 10 mM A – 100 mM Na + 150 mM A – 0.2 mM Cell exterior K + 5 mM Cl – 120 mM Cell exterior Cell interior Plasma membrane Na + –K + pump Diffusion K+K+ Na + Diff us ion -70 mV

36. Seymour Benzer – research leader in studies of Shaker Flies. In the 1970s, his lab was able to associate this mutant’s behavior to a gene that affects potassium channel development.

37. Shaker Potassium Channels Genetic approaches include screening for behavioral changes in animals with mutations in K+ channel genes. Such genetic methods allowed the genetic identification of the "Shaker" K+ channel gene in Drosophila before ion channel gene sequences were well known. The Shaker (Sh) gene, when mutated, causes a variety of atypical behaviors in the fruit fly, Drosophila melanogaster. Under ether anesthesia, the fly’s legs will shake (hence the name); even when the fly is unanaesthetized, it will exhibit aberrant movements. Sh-mutant flies have a shorter lifespan than regular flies; in their larvae, the repetitive firing of action potentials as well as prolonged exposure to neurotransmitters at neuromuscular junctions occurs. The Sh gene plays a part in the operation of potassium ion channels, which are integral membrane proteins and are essential to the correct functioning of the cell.

38. Relative Ion Permeabilities of the Membrane at Rest –K + channels: 4 subunits –Channel selectively permeable to K + ions –MacKinnon—2003 Nobel Prize Mutations of specific K + channels; Inherited neurological disorders

40. END.