Membrane potentials Flashcards
How can glass capillaries achieve intracellular recordings
Microelectrode is filled with a conducting salt solution and a wire inserted into it to connect to an voltmeter that measures the difference in potential between the fine tip (<1um) and a ground
What does penetrating the neuronal membrane with the fine tip of an electrode reveal
A hyperpolarised resting membrane potential (-50 to -90mV, around -65mV)
Why is the plasma membrane called a lipid bilayer
It is made up of phospholipids with hydrophilic heads and hydrophobic tails that line up naturally to form a lipid bilayer when in an aqueous solution (Singer and Nicholson, 1972)
What does the fluid mosaic model describe
The structure of the plasma membrane as a mosaic of components, with cholesterol, proteins and carbohydrates floating in a sea of phospholipids
The phospholipids and embedded proteins can diffuse rapidly and laterally in the membrane
What are the 3 functions of the phospholipid bilayer
Impermeable, proteins allow communication and movement of molecules, insulator
3 functions of the phospholipid bilayer- impermeable
Impermeable to ions and organic molecules- can maintain the cell’s intracellular environment
3 functions of the phospholipid bilayer- proteins allow communicatino and movement of molecuels
The proteins that span from the intracellular to extracellular space allow the cell to communicate with its environment and allow selective movement of molecules across the membrane eg nutrients
3 functions of the phospholipid bilayer- insulator
The bilayer separates ionic charges in the intracellular and extracellular salt fluids, thus acting as an insulator between 2 conductors (capacitor), allowing charge to be stored on the neuronal membrane
What is the consequence of the asymmetric distribution of ions across the cell membrane with an impermeable membrane
Resting membrane potential of 0mV, as the negative and positive charges are balanced across the membrane (despite asymmetric distribution of ions)
Intracellular vs extracellular Na+ conc
Intracellular 18mM
Extracellular 145mM
Intracellular vs extracellular Cl- conc
Intracellular 5mM
Extracellular 115mM
Intracellular vs extracellular Ca2+ conc
Intracellular 100nM
Extracellular 2mM
Intracellular vs extracellular K+ conc
Intracellular 140mM
Extracellular 5mM
Intracellular vs extracelllar organic anions conc
Intracellular 75mM
Extracellular 15mM
What channels can mainly explain the hyperpolarised resting membrane potential
Leak channels in the membrane that are predominantly selective for K+ ions
What is the effect of K+ selective leak channels in the membrane on the movement of K+
K+ ions flow through these channels along their concentration gradient to from the inside to the outside of the cell, and accumulate on the outside surface of the membrane
What is the effect of K+ moving out of selective leak channels
As K+ is positively charged, there is a net accumulation of negative charges on the inner side of the membrane, and the membrane becomes hyperpolarised relative to the extracellular fluid
What is the effect on K+ of the membrane becoming hyperpolarised by movement of K+ out via leak channels
It generates an electrical force that attracts the K+ ions back into the cell
At first the diffusional gradient is stronger than the opposing electrical gradient and there continues to be a net efflux of K+
What happens as the membrane becomes progressively more hyperpolarised by the movement of K+ back into the cell
The system will eventually reach electrochemical equilibrium when the electrical gradient balances the diffusional gradient and there is no net flux of K+ across the membrane
What is the Nernst equation
Calculates the membrane potential at which equilibrium potential is reached by any single ion
How can we test the Nernst equation in real cells
By manipulating extracellular K+ concentration and examining the changes in membrane potential
The membrane potential at each conc should be predicted by the Nernst equation ie proportional to log ([K+]out (Bernstein, 1902)
What is the result of testing the Nernst equation in real cells by manupulating extracellular K+ concentration
Plotting extracellular K+ conc against RMP on a logarithmic scale reveals that it clearly deviates from the Nernst potential for K+ ions at low values of of [K+]out (Hodgkins and Horowicz, 1959)
Why is the RMP of cells not predicted by the Nernst equation for K+
The leak channels are weakly permeable to other ions, mainly Na+ ions moving into the cell (1/100 fold)
Other ions are moving across the membrane that are not taken into account by the Nernst equation for K+ alone
What can be used to calculate the correct electrochemical equilibrium potential for the leak channels (taking into account all ions) aka resting membrane potential
Goldman-Hodgkin-Katz equation- the average of Nernst potentials for each permeable ion weighted by their relative permeabilities
What happens to cations when they dissolve in water
The cations can dissociate as the partial negative charges of the water molecules surround the ion, creating a 3D sphere of water around the ion and allowing it to disperse in solution
What do ions have to shed in order to pass through the pore of the leak channels as single ions
Have to shed their hydration shells
How do ions shed their hydration shells at the pore of the leak channel
At the selectivity filter, the water molecules can be replaced by interactions with oxygen atoms from carbonyl or hydroxyl groups of the protein, literally puttling the K+ ion, allowing it to diffuse through the channel
How are the leak channels 100 fold more permeable to K+ than Na+
At the selectivity filter, the spatial configuration of the oxygen atoms from carbonyl or hydroxyl groups of the protein are better positioned for interactions with the K+ hydration shell, meaning it is harder for Na+ to shed its hydration shell to pass through the channel, enabling selectivity
What does setting up and maintaining ion gradients across the membrane depend on
Active transport to move ions up their electrochemical gradients
What does primary active transport use energy from
Hydrolysis of ATP
What are active transporters called
Pumps
What is the primary example of a membrane pump
The Na+ pump, which simultaneously pumps 3 Na+ ions out and 2 K+ ions in, using energy from hydrolysis of ATP
What does it mean to say the Na+/K+ pump is electrogenic
It directly causes a small hyperpolarisation of the membrane (few mV) due to imbalances in the movement of charge (net loss of one + ion)
What is secondary active trasport
When the energy stored in the electrochemical gradient is used to drive active transport aka movement of the ion up their concentration gradient is coupled with the movement of another ion down their concentration gradient
What are antiports
Ion exchangers- the driving ion and driven ion/molecule move in opposite directions
What is an example of an antiport
eg Na+/Ca2+ exchanger where the flow of Na+ down its electrochemical gradient back into the cell is used to expel Ca2+
What are symports
Cotransporters- the direction of transport is the same for the driving ion and the driven ion/molecule
What is example of a symport
K+/Cl cotransporter, where both K+ and Cl- move out of the cell
What are types of secondary active transport proteins
Symports and antiports
Evidence that the negative resting membrane potential of neurons is mainly due to leak channels not active transport
If we take a quiescent isolated neuron and block Na+/K+ pump using oubain, there is a small depolarisation due to the lost electrogenic activity of the pump, but the resting membrane potential does not collapse (Brisson et al, 2014)
How does oubain affect active neuronal networks differently to isolated neurons
In active neuronal networks, much higher demands for transporters to maintain intracelllar/extracellular environments so applying oubain can lead to rapid collapse of membrane potentials
eg glia use active transport to maintain ion gradients and uptake neurotransmitters, active transport used for neuronal signalling
What is the process of osmosis
If a membrane is semi-permeable to water, the water molecules will move down their concentration gradient from a region of low solute concentration to higher solute concentration
What is the effect on the water solution when a solid is placed into a chamber on one side of a semi-permeable membrane
The solid dissolves and becomes surrounded by hydration shells, leaving less free water molecules on this side
Water moves by osmosis down its conc gradient to this side, exerting hydrostatic pressure on the membrane that tries to push the water molecules back across the membrane
When does the system reach equilibrium in a chamber with a semi-permeable membrane where a solid is dissolved on one side
The system reaches equilibrium when the hydrostatic pressure balances the osmotic pressure
What impact can osmotic pressure on the membrane have on cells
Can cause them to shrivel or swell
What can cause neuron swelling
Accumulation of ions inside the neuron leads to the influx of water, and the corresponding increase in hydrostatic pressure will cause the neuron to swell
What can be the effect of cellular swelling
Reduces the volume of the extracellular space which affects the conc and diffusion of molecules in the extracellular fluid, which alters neuronal excitability
When may cellular swelling occur
May occur to some extent during periods of high electrical activity, but is more prononuced under pathological conditions eg ischemic stroke (loss of blood supply, leading to oxygen and glucose deprivation)
What normal long term processes involve changes in neuronal volume
Physiological processes like growth, endocytosis, endocytosis
What are aquaporins
Specialised water channels in the membrane that facilitate osmosis
Study showing the importance of aquaporins
Deleting the gene for Aquaporin4 in mice is associated with a sevenfold reduction in cell plasma membrane water permeability
Howevever, these mice still appear largely normal, so slow mechanisms of water transport appear sufficient to sustain normal brain function
Modelling the neuronal membrane as an electronic circuit- what are the leak channels represented as
A resistor, with the ability to conduct current specified in terms of resistance or conductance
If conductance is high, ions can move very rapidly through the pore and vv
Modelling the neuronal membrane as an electronic circuit- what are the ionic gradients across the membrane represented as
A battery (chemical driving force) in series with the resistor, which tries to clamp the membrane circuit at the Nernst potential of the channels
Modelling the neuronal membrane as an electronic circuit- what does the circuit give a transmembrane potential of
-60mV, with current continuously flowing through the leak conductance
Modelling the neuronal membrane as an electronic circuit- what can predict the change in transmembrane potential when current is injected into the circuit
The change in transmembrane potential can be predicted by Ohm’s law
Modelling the neuronal membrane as an electronic circuit- what property of the cells causes the time delay in reaching the membrane potential predicted by Ohm’s law
Due to the capacitance of the membrane (consists of an insulator separating 2 electrical conductive surfaces which can be modelled as a capacitor in parallel with the membrane conductance