Module 5: Membrane Properties

Topic 1: Cell Surface Charge

1. Introduction
 All living cells carry electrical charges on their surface.
 This arises from:
o Charged membrane lipids (e.g., phosphatidylserine, phosphatidylinositol, gangliosides).
o Membrane proteins (amino acid side chains with carboxylate, amino, or phosphate groups).
o Carbohydrate moieties (glycoproteins, glycolipids → sialic acid residues are negatively
charged).
Net effect: The cell surface usually carries a negative charge under physiological conditions.

2. Sources of Cell Surface Charge

2.1 Lipids
 Phospholipids (asymmetric distribution):
o Outer leaflet: phosphatidylcholine, sphingomyelin (neutral).
o Inner leaflet: phosphatidylserine, phosphatidylinositol (negatively charged).
 Negatively charged lipids → contribute to overall surface charge.
2.2 Proteins
 Membrane proteins have ionizable groups (–COO⁻, –NH₃⁺).
 Surface proteins with acidic amino acids (Asp, Glu) increase negative charge.
2.3 Carbohydrates
 Glycocalyx (sugar coat) rich in sialic acid residues → strongly negative.
 Important for: cell–cell recognition, adhesion, immune interactions.

3. Measurement of Surface Charge

 Zeta potential:
o The electrical potential at the shear plane of a moving cell in solution.
o Negative in most mammalian cells (−10 to −30 mV).
 Electrophoretic mobility: Cells move toward the anode in an electric field due to negative surface
charge.
4. Functional Significance
1. Cell–Cell Interaction
o Negative charges prevent uncontrolled cell aggregation (electrostatic repulsion).
o Selective adhesion mediated by Ca²⁺ bridges or specific adhesion molecules.
2. Ion Binding & Transport
o Negative charges attract cations (Ca²⁺, Mg²⁺, Na⁺).
o Regulate local ion concentrations near the membrane.
3. Immune Recognition
o Altered surface charge can signal apoptosis (phosphatidylserine externalization).
o Pathogens often exploit glycocalyx for binding/entry.
4. Drug & Nanoparticle Interaction
o Cell surface charge affects uptake of charged molecules and nanoparticles.
o Positively charged drug carriers bind more effectively to negatively charged cell membranes.
5. Electrical Properties
o Surface charges influence electric double layer and thereby local field effects on ion
channels and receptors.

5. Clinical & Biological Relevance

 Cancer cells: Often have altered surface charge (different sialylation patterns).
 Apoptosis: “Eat-me” signal = externalized phosphatidylserine → macrophage recognition.
 Infection: Viruses/bacteria interact with glycocalyx charges (e.g., influenza binds sialic acid).
 Blood compatibility: Surface charge of RBCs (negative due to sialic acid) prevents
clotting/aggregation.

6. Quick Recap Table

Source Contribution to Surface Charge Importance

Lipids PS, PI (negative head groups) Apoptosis signaling, ion binding

Proteins Ionizable amino acids Receptor & enzyme activity

Carbohydrates Sialic acid in glycocalyx Adhesion, immune recognition

Zeta potential Measure of surface charge Diagnostic/biophysical tool

Key Concept:
Cell surface charge is mainly negative, arising from lipids, proteins, and glycocalyx. It influences cell–cell
interactions, ion binding, immune recognition, and drug delivery.
Topic 2: Resting Membrane Potential

1. Definition
 Resting Membrane Potential (RMP): The steady voltage difference across the cell membrane
when the cell is at rest (not excited).
 Typical values:
o Neurons: −60 to −70 mV
o Skeletal muscle: −85 to −90 mV
o Cardiac cells: −80 mV
o Plant cells: up to −120 mV
Always negative inside relative to outside.

2. Basis of RMP
2.1 Ion Gradients (Established by Pumps)
 Na⁺/K⁺ ATPase:
o Pumps 3 Na⁺ out, 2 K⁺ in.
o Maintains high K⁺ inside (~140 mM) & high Na⁺ outside (~145 mM).
o Electrogenic: contributes a few mV negativity.
 Other pumps: Ca²⁺ ATPase, H⁺ ATPase (plants/fungi).
2.2 Selective Permeability
 K⁺ leak channels → dominant at rest.
 Na⁺ & Cl⁻ permeabilities are small but significant.
 Therefore, Vm ≈ E_K but slightly depolarized.
2.3 Impermeant Anions
 Intracellular proteins, nucleic acids contribute negative charge.
 Donnan effect influences ion distribution.

3. Quantitative Framework
3.1 Nernst Equation (single ion equilibrium)

RT [ ion ] outside
Eion = ln ⁡( )
zF [ ion ]inside
 Example:
o For K⁺: E_K ≈ −90 mV.
o For Na⁺: E_Na ≈ +60 mV.
3.2 Goldman–Hodgkin–Katz (GHK) Equation
V m =61mV ⋅log ¿ ¿

 Accounts for relative permeabilities (P).

 Explains why Vm ≈ −70 mV, not exactly −90 mV (pure E_K).

4. Membrane Properties & RMP

 Capacitance (C_m): ~1 µF/cm² → determines how fast Vm can change.
 Resistance (R_m): High at rest → few leak channels open.
 Time constant (τ = R_m × C_m): Defines how quickly Vm responds to inputs.
 Membrane proteins:
o Ion channels = pathways for passive flux.
o Pumps = maintain gradients (active processes).

5. Physiological Significance
 Electrical excitability: Foundation for action potentials in nerves & muscle.
 Transport processes: Secondary active transport relies on RMP (e.g., Na⁺-driven glucose uptake).
 Osmotic balance: RMP helps counteract Donnan-driven swelling.
 Synaptic integration: PSPs depolarize or hyperpolarize relative to RMP.

6. Clinical Relevance
 Hyperkalemia (↑[K⁺]out):
o Makes RMP less negative (depolarization).
o Can inactivate Na⁺ channels → cardiac arrhythmias.
 Hypokalemia (↓[K⁺]out):
o Makes RMP more negative (hyperpolarization).
o Leads to muscle weakness, paralysis.
 Channelopathies: Mutations in leak channels alter RMP stability.
 Cardiac glycosides (e.g., digoxin): Inhibit Na⁺/K⁺ ATPase → depolarize RMP → ↑contractility.

7. Quick Recap Table

Factor Contribution to RMP

Na⁺/K⁺ ATPase Establishes gradients, electrogenic (−5 to −10 mV)

K⁺ leak channels Major determinant (Vm ≈ E_K)

Factor Contribution to RMP

Na⁺ leak channels Slight depolarizing effect

Cl⁻ equilibrium Stabilizes Vm

Impermeant anions (Pr⁻) Donnan effect, osmotic balance

Key Concept:
The resting membrane potential is mainly determined by K⁺ diffusion through leak channels, but fine-
tuned by Na⁺, Cl⁻, and electrogenic pumps. It provides the baseline electrical state necessary for
excitability, signaling, and transport.
Topic 3: Action Potential & Properties

1. Definition
 An Action Potential (AP) is a rapid, transient, regenerative electrical event in excitable membranes.
 Functions:
o Long-distance communication (neurons).
o Triggering contraction (muscle).
o Coordinating rhythmic activity (cardiac pacemaker cells).

2. Ionic Basis of Action Potential

1. Resting state
o Vm ≈ −70 mV.
o Dominant permeability: K⁺ leak channels.
2. Depolarization (Rising phase)
o Stimulus brings Vm to threshold (~ −55 mV).
o Voltage-gated Na⁺ channels open → Na⁺ influx.
o Positive feedback → rapid upstroke.
3. Overshoot
o Vm becomes positive (~ +30 mV).
o Driving force on Na⁺ decreases.
4. Repolarization (Falling phase)
o Na⁺ channels inactivate.
o Voltage-gated K⁺ channels open → K⁺ efflux.
o Vm moves back toward E_K.
5. Afterhyperpolarization (Undershoot)
o Vm goes slightly more negative than rest due to prolonged K⁺ conductance.
6. Recovery
o K⁺ channels close, Vm returns to resting potential.
o Na⁺/K⁺ ATPase restores ionic gradients.

3. Properties of Action Potential

3.1 All-or-None Principle
 If stimulus ≥ threshold → full AP.
  Subthreshold → no AP.
 Suprathreshold → same amplitude AP (not bigger).
3.2 Threshold
 The critical level of depolarization required to trigger Na⁺ channel opening.
3.3 Refractory Periods
 Absolute refractory period:
o Na⁺ channels inactivated → no AP possible.
 Relative refractory period:
o K⁺ channels still open → stronger stimulus needed.
 Ensures unidirectional propagation.
3.4 Conduction Properties
 Non-decremental propagation: AP amplitude does not decay along axon.
 Conduction velocity depends on:
o Axon diameter (larger = faster).
o Myelination (saltatory conduction).
o Membrane capacitance & resistance (affect time constant, space constant).
3.5 Stereotyped Waveform
 AP shape (amplitude, duration) is consistent for a given cell type.

4. Membrane Properties & AP

 Capacitance (C_m):
o Determines how quickly Vm can change.
o Lower C_m (as in myelinated axons) → faster conduction.
 Resistance (R_m):
o High R_m (myelination) reduces leak → longer length constant (λ).
o AP spreads further before decaying, allowing saltatory conduction.
 Time constant (τ = R_m × C_m):
o Short τ → faster depolarization.
o Long τ → more temporal summation of inputs.

5. Functional Importance
 Neurons: Basis for information coding (frequency, pattern of APs).
 Skeletal muscle: AP triggers Ca²⁺ release → contraction.
  Cardiac muscle: AP ensures rhythmic contraction & refractory period prevents tetanus.
 Endocrine cells: APs trigger hormone secretion (e.g., insulin release).

6. Clinical Relevance
 Na⁺ channel blockers (lidocaine, TTX): Prevent AP initiation → anesthesia.
 K⁺ channel blockers (dendrotoxin): Prolong AP → hyperexcitability.
 Demyelination (MS, Guillain–Barré): Slows conduction → conduction block.
 Electrolyte imbalance:
o Hyperkalemia → depolarizes RMP, inactivates Na⁺ channels.
o Hypokalemia → hyperpolarizes RMP, harder to reach threshold.

7. Quick Recap Table

Property Feature

All-or-none AP fires only if threshold reached

Threshold Critical depolarization (~ −55 mV)

Refractory periods Absolute (no AP), Relative (needs strong stimulus)

Conduction Active, regenerative, non-decremental

Velocity factors Diameter ↑, Myelination ↑, Capacitance ↓

Na⁺ influx → depolarization; K⁺ efflux →

Ionic basis
repolarization

Key Concept:
Action potentials are regenerative, all-or-none signals shaped by Na⁺ and K⁺ channel dynamics and
strongly influenced by membrane capacitance, resistance, and myelination.
Topic 4: Permeability Changes during Action Potential

1. Introduction
 The action potential (AP) is driven by time- and voltage-dependent changes in membrane
permeability (P) to different ions.
 At rest: high P_K, low P_Na, moderate P_Cl.
 During AP: dynamic shifts in P_Na and P_K underlie depolarization and repolarization.
- First revealed by Hodgkin & Huxley (1952) using voltage clamp experiments in squid giant axon.

2. Permeability Sequence During AP

2.1 Resting State
 High P_K (K⁺ leak channels open).  Vm close to E_K (~ −70 mV).
 Very low P_Na.

2.2 Depolarization Phase (Upstroke)

 Threshold depolarization → opens  Massive Na⁺ influx drives Vm toward
voltage-gated Na⁺ channels. E_Na (+60 mV).
 P_Na increases ~1000-fold in <1 ms.  Positive feedback: depolarization opens
more Na⁺ channels.

2.3 Overshoot
 Vm becomes positive (~ +30 mV).  Na⁺ channels begin to inactivate.
 Driving force for Na⁺ decreases as Vm
approaches E_Na.

2.4 Repolarization Phase

 P_Na decreases as Na⁺ channels  P_K increases 10–20× above resting.
inactivate.
 Strong K⁺ efflux repolarizes Vm toward
 Voltage-gated K⁺ channels open E_K.
(delayed rectifiers).

2.5 Afterhyperpolarization (Undershoot)

 K⁺ channels remain open longer than Na⁺ channels.
 P_K still elevated → Vm goes below resting (closer to E_K).
 Eventually K⁺ channels close → Vm returns to RMP.
3. Graphical Summary
 If you plot relative permeability (P_Na, P_K) vs. time:
o P_Na: rapid spike during depolarization, then falls due to inactivation.
o P_K: slower rise, peaks during repolarization, remains high during undershoot, then returns
to baseline.

4. Functional Importance
 Unidirectional propagation: Na⁺ channel inactivation + refractory period ensures AP moves
forward.
 Repolarization speed: K⁺ conductance restores Vm rapidly, readying cell for next AP.
 Excitability control: Balance of Na⁺ and K⁺ permeability sets threshold and firing frequency.

5. Clinical & Pharmacological Relevance

 Tetrodotoxin (TTX): Blocks Na⁺ channels → prevents AP (no depolarization).
 Local anesthetics (lidocaine): Reversibly block Na⁺ channels → pain blockade.
 K⁺ channel blockers (4-aminopyridine, dendrotoxin): Prolong AP → used experimentally and in
MS therapy (to enhance conduction).
 Hyperkalemia/hypokalemia: Alter driving force for K⁺, affecting repolarization and excitability.

6. Quick Recap Table

Phase P_Na P_K Result

Resting Low High Vm ≈ E_K

Depolarization ↑↑↑ (fast) Low Na⁺ influx, Vm → E_Na

Overshoot Falling (inactivation) Starting to rise Vm positive

Repolarization Low (inactivated) ↑↑↑ (delayed) K⁺ efflux, Vm → E_K

Afterhyperpolarization Low Still high Vm below RMP

Recovery Low Resting level Vm returns to RMP

Key Concept:
The action potential is produced by a transient, sequential rise and fall in Na⁺ and K⁺ permeability:
 Na⁺ permeability dominates early → depolarization.
 K⁺ permeability dominates later → repolarization & undershoot.
Topic 5: Ion Channels

1. Introduction
 Ion channels are transmembrane proteins that form selective pores for ion movement.
 Conduct ions at very high rates (10⁶–10⁸ ions/sec) compared to transporters.
 Provide selectivity, gating (open/close control), and fast signaling.
 Essential for resting membrane potential, action potentials, synaptic transmission, secretion,
and sensory perception.

2. General Structure of Ion Channels

 Subunit organization: Many channels are oligomeric (tetramers, pentamers, or heteromultimers).
 Pore region: Forms ion conduction pathway.
 Selectivity filter: Determines which ions pass (size, charge, hydration energy).
 Gating domains: Sense stimuli and control open/closed states.
 Accessory subunits: Modulate channel properties (kinetics, localization).

3. Types of Ion Channels

3.1 By Selectivity
 K⁺ channels: Highly selective for K⁺ over Na⁺ (>1000:1).
 Na⁺ channels: Selective for Na⁺, critical for AP initiation.
 Ca²⁺ channels: Selective for Ca²⁺, key in neurotransmitter release & contraction.
 Cl⁻ channels: Allow anion flow, regulate excitability and cell volume.
 Non-selective cation channels: Permit multiple cations (e.g., NMDA receptor).

3.2 By Gating Mechanism

1. Voltage-gated channels (VGICs)
o Open in response to changes in membrane potential.
o Families: Naᵥ, Kᵥ, Caᵥ, Clᵥ.
o Example: VG Na⁺ channels in axons → AP depolarization.
2. Ligand-gated ion channels (LGICs / ionotropic receptors)
o Open when neurotransmitter/ligand binds.
o Example: Nicotinic ACh receptor (cation channel), GABA_A receptor (Cl⁻).
3. Mechanosensitive channels
o Open in response to stretch, pressure, or vibration.
 o Example: Hair cell channels in cochlea (hearing).
4. Second messenger–gated channels
o Regulated by intracellular messengers (cAMP, IP₃, Ca²⁺).
o Example: cAMP-gated channels in olfactory neurons.
5. Leak channels (background channels)
o Always open, contribute to RMP.
o Example: K2P K⁺ leak channels.

4. Ion Channel Selectivity

 Determined by pore size, charge distribution, and hydration shell interactions.
 Example: K⁺ channels exclude Na⁺ even though Na⁺ is smaller → filter stabilizes dehydrated K⁺
optimally, but not Na⁺.

5. Functional Roles
 Resting potential: K⁺ leak channels.
 Action potentials: Voltage-gated Na⁺ and K⁺ channels.
 Synaptic transmission: Ligand-gated channels (ionotropic glutamate, GABA_A).
 Excitation–contraction coupling: Ca²⁺ channels in muscle.
 Secretion: Ca²⁺ entry → exocytosis of neurotransmitters/hormones.
 Sensory functions: Mechanosensitive & cyclic nucleotide-gated channels in touch, smell, vision,
hearing.

6. Channelopathies (Diseases of Ion Channels)

 Na⁺ channelopathies: Epilepsy, periodic paralysis, cardiac arrhythmias.
 K⁺ channelopathies: Long QT syndrome, ataxia, epilepsy.
 Ca²⁺ channelopathies: Migraine, congenital night blindness, Lambert-Eaton syndrome.
 Cl⁻ channelopathies: Cystic fibrosis (CFTR mutation), myotonia congenita.
 Mechanosensitive channel defects: Contribute to kidney disorders, hypertension.

7. Experimental Methods
 Patch-clamp recording: Measures single-channel currents.
 Voltage clamp: Reveals ionic permeability changes during AP.
 Cryo-EM/X-ray crystallography: Determines atomic structures.
 Molecular genetics: Identify mutations underlying channelopathies.
8. Clinical & Pharmacological Importance
 Local anesthetics (lidocaine, procaine): Block voltage-gated Na⁺ channels.
 Anti-epileptics (phenytoin, carbamazepine): Modulate Na⁺/Ca²⁺ channels.
 Anti-arrhythmics: Target Na⁺ and K⁺ channels.
 Benzodiazepines: Enhance GABA_A receptor channel opening.
 CF therapy: Drugs (ivacaftor) that restore CFTR Cl⁻ channel function.

9. Quick Recap Table

Channel Type Gating Mechanism Selectivity Example Function

Na⁺, K⁺, Ca²⁺,

Voltage-gated Membrane potential Naᵥ in axons AP generation
Cl⁻

Ligand-gated Neurotransmitters Cations/Cl⁻ nAChR, GABA_A Synaptic transmission

Mechanosensitive Stretch/pressure Non-specific Hair cell channels Hearing, touch

cAMP-gated
Second messenger-gated cAMP, IP₃, Ca²⁺ Cations Sensory pathways
channels

Leak channels Always open K⁺ (mostly) K2P Resting potential

Key Concept:
Ion channels are selective, gated pores essential for electrical excitability, signaling, and homeostasis. Their
dysfunction leads to channelopathies, and they are major drug targets.
Topic 6: Cardiac Muscle Action Potential

1. Introduction
 Cardiac muscle APs are longer in duration (200–400 ms) than neuronal APs (~2 ms).
 This ensures sustained contraction (systole) and prevents tetany.
 Different cardiac cells have distinct AP shapes:
o Ventricular/atrial myocytes → long plateau-type AP.
o Pacemaker cells (SA & AV nodes) → slow, spontaneous depolarization (automaticity).

2. Ventricular Myocyte Action Potential (Phases 0–4)

Phase 0: Rapid Depolarization
 Triggered when Vm reaches threshold (~ −70 mV).
 Fast Na⁺ channels open → Na⁺ influx.
 Vm rises to +20 to +30 mV.
Phase 1: Initial Repolarization
 Na⁺ channels inactivate.
 Transient outward K⁺ current (I_to) → small repolarization.
Phase 2: Plateau Phase (unique to cardiac AP)
 Balance between:
o Ca²⁺ influx (via L-type Ca²⁺ channels, I_Ca-L).
o K⁺ efflux (delayed rectifier K⁺ channels).
 Maintains Vm near 0 mV for ~200 ms.
 Function: Allows Ca²⁺ entry → triggers Ca²⁺ release from SR → contraction.
Phase 3: Repolarization
 Ca²⁺ channels close.
 K⁺ efflux dominates (delayed rectifiers, I_K).
 Vm returns toward E_K (~ −90 mV).
Phase 4: Resting Membrane Potential
 Maintained by K⁺ leak channels (I_K1).
 Na⁺/K⁺ ATPase restores gradients.
 Stable in ventricular myocytes (not in pacemaker cells).

3. Pacemaker Cell Action Potential (SA & AV nodes)

 No stable RMP → instead, a slow depolarization (pacemaker potential) in Phase 4.
  Ionic basis:
o Funny current (I_f): Na⁺ “leak” inward during hyperpolarization.
o T-type Ca²⁺ channels contribute near threshold.
o L-type Ca²⁺ channels cause Phase 0 depolarization (instead of fast Na⁺).
o Repolarization (Phase 3) by K⁺ efflux.
 Responsible for automaticity of the heart.

4. Refractory Periods
 Absolute refractory period (ARP): During plateau & early repolarization → no new AP possible.
 Relative refractory period (RRP): During late repolarization → stronger stimulus may trigger AP.
 Long refractory period prevents tetanic contraction → ensures rhythmic beating.

5. Functional Importance
 Plateau phase: Ensures long contraction → effective ejection of blood.
 Ca²⁺ influx: Couples excitation to contraction (excitation–contraction coupling).
 Refractoriness: Prevents arrhythmias and maintains rhythmic pumping.

6. Clinical Relevance
 Anti-arrhythmic drugs:
o Class I (Na⁺ channel blockers): slow Phase 0 depolarization.
o Class II (β-blockers): reduce Ca²⁺ currents, slow conduction.
o Class III (K⁺ channel blockers): prolong repolarization & refractory period.
o Class IV (Ca²⁺ channel blockers): reduce plateau phase & excitability.
 Ischemia: Reduces ATP → impairs Na⁺/K⁺ pump → depolarization → arrhythmias.
 Long QT syndrome: Delayed repolarization (Phase 3) → risk of torsades de pointes.
 Digitalis (digoxin): Inhibits Na⁺/K⁺ ATPase → ↑ intracellular Ca²⁺ → ↑ contractility.

7. Quick Recap Table

Phase Vm Change Ion Movements Notes

0 Rapid depolarization Na⁺ influx (fast Na⁺ channels) Upstroke

1 Initial repolarization K⁺ efflux (I_to) Brief dip

Ca²⁺ influx (L-type) ≈ K⁺

2 Plateau Excitation–contraction coupling
efflux
Phase Vm Change Ion Movements Notes

3 Repolarization K⁺ efflux (delayed rectifiers) Vm → E_K

4 Resting potential K⁺ leak, Na⁺/K⁺ ATPase Stable in myocytes

Key Concept:
The cardiac action potential is longer and more complex than neuronal APs due to the plateau phase
(Ca²⁺ influx). This ensures sustained contraction, refractory period, and rhythmic pumping of the
heart.
Topic 7: Nernst & Goldman Equations

1. Nernst Equation
1.1 Purpose
 Describes the equilibrium potential (Eion) for a single ion species across a membrane.
 Predicts the voltage at which the electrochemical driving force is zero (no net ion flux).

1.2 Formula
RT [ion]outside
Eion = ln ( )
zF [ion ]inside

At 37 °C (310 K), simplified:

E K =61mV ⋅log ¿ ¿

where:
 R = gas constant = 8.314 J·mol⁻¹·K⁻¹
 T = absolute temperature (K)
 F = Faraday constant = 96,485 C/mol
 z = ion valence
 [[ion] outside, [ion]inside = extracellular and intracellular concentrations

1.3 Example (Neuron, K⁺)

 ¿¿
5
E K =61⋅ log ⁡( )≈−90 mV
140
 Thus, if Vm ≠ −90 mV, K⁺ will flow to drive Vm toward E K .

2. Goldman–Hodgkin–Katz (GHK) Equation

2.1 Purpose
 Extends Nernst → accounts for multiple ions and their relative permeabilities (P).
 Explains the resting membrane potential (Vm) in real cells.

2.2 Formula
Vm=61 mV ⋅ log¿ ¿
Note: For Cl⁻ (anion), the inside/outside terms are flipped because of its negative charge.
2.3 Key Insights
 At rest, PK≫PNa,PCl → Vm is close to EK (−90 mV).
 But because Na⁺ permeability is nonzero, Vm is slightly depolarized (~ −70 mV).
 Cl⁻ acts as a stabilizer, balancing charges.

3. Differences Between Nernst & Goldman

Feature Nernst Equation Goldman Equation

Ions considered Single ion Multiple ions

Condition Equilibrium (net ion flux = 0) Steady-state (net current = 0, ions still moving)

Predicts Reversal potential (Eion) Resting potential (Vm)

4. Physiological Significance
 Nernst:
o Predicts reversal potential of ion-selective channels.
o Determines ion flux direction at synapses or AP phases.
 Goldman:
o Explains resting potential (Vm ≈ −70 mV in neurons).
o Basis for understanding how changes in extracellular [K⁺], [Na⁺], [Cl⁻] alter excitability.

5. Clinical Relevance
 Hyperkalemia: ↑[K⁺]out → E_K less negative → depolarized Vm → arrhythmia risk.
 Hypokalemia: ↓[K⁺]out → E_K more negative → hyperpolarized Vm → muscle weakness.
 Ischemia: Na⁺/K⁺ ATPase failure → gradients collapse → Vm drifts toward 0.
 Drug targets: Many anesthetics and antiepileptics alter ion permeabilities → shift Vm.

6. Quick Recap Table

Equation Expression Application

61 [out]
Nernst Eion = log ( ) Single-ion equilibrium potential
z [¿]

Goldman Vm=61 mV ⋅ log¿ ¿ Multi-ion resting potential

Key Concept:
 Nernst equation → “equilibrium potential” for one ion.
 Goldman equation → “resting potential” considering all permeant ions.
Topic 8: Hodgkin–Katz Experiments

1. Historical Context
 1939–1949: Alan Hodgkin & Bernard Katz studied the squid giant axon (~1 mm diameter).
 Goal: Understand ionic basis of the resting membrane potential (RMP).
 Why squid axon? → Large size made it possible to insert electrodes and measure intracellular
potentials.

2. Experimental Setup
 Inserted microelectrodes inside axon to measure membrane potential (Vm).
 Manipulated external ion concentrations ([K⁺], [Na⁺], [Cl⁻]).
 Compared measured Vm with predictions from Nernst equation.

3. Key Observations
1. Potassium dependence
o Changing extracellular [K⁺] shifted Vm in a way predicted by the Nernst equation for K⁺.
o Strong evidence that K⁺ is the main determinant of RMP.
2. Role of sodium
o Vm was not exactly equal to E_K (−90 mV).
o Instead, Vm was closer to −65 mV.
o Concluded: Na⁺ permeability is small but nonzero at rest, pulling Vm slightly positive.
3. Role of chloride
o Cl⁻ passively distributes near equilibrium, also influences Vm.
4. Na⁺/K⁺ Pump
o Showed that RMP is a steady-state, not a true equilibrium.
o Pump maintains gradients (3 Na⁺ out, 2 K⁺ in).

4. Development of the Hodgkin–Katz Equation

 Generalized Goldman’s constant field equation by adding permeability terms.
 Final form (Goldman–Hodgkin–Katz, GHK equation):
Vm=61 mV ⋅ log¿ ¿
 This equation explained the measured Vm (~ −65 mV) far better than the K⁺-only Nernst potential.

5. Significance
  Experimental proof that RMP is determined by multiple ions (mainly K⁺, with smaller Na⁺ & Cl⁻
contributions).
 Showed that Vm is a steady-state potential maintained by ion pumps, not a passive equilibrium.
 Formed the foundation for Hodgkin–Huxley model (1952) → ionic basis of the action potential.

6. Clinical & Physiological Importance

 Explains how [K⁺]out changes alter excitability:
o Hyperkalemia: Depolarizes Vm → arrhythmias.
o Hypokalemia: Hyperpolarizes Vm → muscle weakness.
 Understanding permeability ratios helps in designing drugs that target ion channels (anti-
arrhythmics, anesthetics).
 Basis for electrophysiological experiments (voltage clamp, patch clamp).

7. Quick Recap Table

Observation Hodgkin–Katz Conclusion

Vm shifted with [K⁺]out RMP mainly set by K⁺ gradient

Vm not equal to E_K Na⁺ contributes slightly

Cl⁻ distribution Helps stabilize Vm

Pump activity required RMP is a steady state, not equilibrium

Equation developed GHK equation predicts Vm (~ −65 mV)

Key Concept:
The Hodgkin–Katz experiments proved that the resting membrane potential depends not just on K⁺, but
on the combined permeabilities of K⁺, Na⁺, and Cl⁻, maintained by active ion pumps.
Topic 9: Role of Potassium (K⁺) and Sodium (Na⁺)

1. Introduction
 K⁺ and Na⁺ are the principal cations in excitable cells.
 Their unequal distribution across the plasma membrane underlies:
o Resting membrane potential (RMP).
o Action potential generation & propagation.
o Secondary transport processes.
 Gradients maintained by Na⁺/K⁺ ATPase.

2. Distribution of K⁺ and Na⁺ in Mammalian Neurons

Ion Intracellular [ ] Extracellular [ ] Ratio

K⁺ ~140 mM ~5 mM 28 : 1 (inside > outside)

Na⁺ ~10–15 mM ~145 mM 1 : 14 (outside > inside)

These steep gradients are essential for electrical excitability.

3. Role of K⁺ (Potassium)
1. Resting Membrane Potential
o Dominant contributor (via K⁺ leak channels).
o RMP close to E_K (−90 mV).
2. Repolarization of Action Potential
o Voltage-gated K⁺ channels open during AP → K⁺ efflux.
o Brings Vm back toward E_K.
3. Afterhyperpolarization
o Delayed K⁺ channel closing → Vm temporarily below RMP.
4. Cell Volume Regulation
o K⁺ efflux balances osmotic effects of intracellular anions.
5. Excitability Control
o [K⁺]out strongly influences excitability:
 ↑[K⁺]out (hyperkalemia) → depolarization → Na⁺ channel inactivation →
arrhythmias.
 ↓[K⁺]out (hypokalemia) → hyperpolarization → decreased excitability.

4. Role of Na⁺ (Sodium)

 1. Depolarization of Action Potential
o Voltage-gated Na⁺ channels open rapidly at threshold.
o Massive Na⁺ influx drives Vm toward E_Na (+60 mV).
2. Driving Force for Secondary Active Transport
o Na⁺ gradient powers symporters/antiporters (e.g., Na⁺/glucose, Na⁺/Ca²⁺ exchanger).
3. Excitation–Secretion Coupling
o Na⁺ entry depolarizes membrane → activates voltage-gated Ca²⁺ channels →
neurotransmitter/hormone release.
4. Recovery & Gradient Maintenance
o Na⁺/K⁺ ATPase extrudes Na⁺, preventing osmotic swelling.

5. Role of Na⁺/K⁺ ATPase

 Pumps 3 Na⁺ out, 2 K⁺ in per ATP.
 Maintains gradients critical for RMP & excitability.
 Electrogenic: contributes −5 to −10 mV to RMP.
 Inhibited by ouabain/digoxin → depolarization, altered excitability, ↑contractility (heart).

6. Interplay of K⁺ and Na⁺ in Membrane Potentials

 Resting potential: Mainly set by K⁺ (due to leak channels) but shifted toward Na⁺ because of small
Na⁺p permeability.
 Action potential:
o Rising phase → Na⁺ influx. o Falling phase → K⁺ efflux.
 Excitability: Depends on relative balance of Na⁺ and K⁺ driving forces.

7. Clinical Relevance
 Hyperkalemia (↑K⁺out):
o Depolarizes Vm, reduces Na⁺ channel availability → arrhythmias, muscle weakness.
 Hypokalemia (↓K⁺out):
o Hyperpolarizes Vm → reduced excitability → paralysis.
 Hyponatremia/Hypernatremia:
o Alter osmotic gradients → cerebral edema or shrinkage.
 Na⁺ channel blockers (lidocaine, TTX): Prevent AP initiation → anesthetics, toxins.
 K⁺ channel blockers (4-AP, dendrotoxin): Prolong AP → potential therapy for MS.
8. Quick Recap Table

Ion Major Role at Rest Role in AP Other Functions

K⁺ Sets RMP (via leak) Repolarization, afterhyperpolarization Volume regulation, excitability control

Na⁺ Minor at RMP Depolarization (rising phase) Secondary transport, secretion, excitability

Key Concept:
 K⁺ dominates resting potential & repolarization.
 Na⁺ dominates depolarization & drives secondary transport.
 Together, regulated by Na⁺/K⁺ ATPase, they form the core ionic basis of excitability and
homeostasis.