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• 19th century Galvanism demonstrated by Luigi & Lucia Galvani. They investigated role of
electricity in nerves by using two probes made of different metals (copper and iron)
• Galvanism: Contraction of a muscle that is stimulated by an electric current.
Galvani believed in idea called animal electricity, however it was proven incorrect
• Animal electricity: Vital fluid that produces movement in animals.
• Allessandro Volta argued against Galvanism by refuting animal electricity, and claiming only
one type of electricity exists. He believed electricity is generated chemically.
Voltaic pile: Stacks of alternating metals (electrodes) soaked in electrolytes.
• Galvanism (animal electricity) V. Volta (regular electricity) debate
Bioelectricity
• Giovani Aldini stimulated corpse with electricity allowing reanimation (movement, sitting up)
• Vitalist (Unique laws apply to living beings) v. Mechanist (Universal laws apply to everything)
• Aldini’s ideas lead to understanding of electricity in movement, and heart
• Camillo Golgi (1873) developed the black reaction to prove the reticular theory
• Black reaction: Block of formaldehyde fixed tissue submerged into potassium dichromate
for 2 days (hardens tissue), then immersion in silver nitrate (silver chromate salt) for 2 days.
Silver forms black precipitate which fills entire neuron to allow visualization of shape
Only stains 5% of neurons in tissue which shows patches of staining
Neuron Doctrine v. Reticular Theory
Localization of Function
• Localization of function: Brain has discrete regions that serve specific functions
Developed during the 19th & 20th century
• Dr. Paul Broca studied a patient with expressive aphasia (only says one word). Broca
dissected brain post-mortem and searched for lesions. Large lesion in Broca’s area
First demonstration of localization of function in brain
• Greek physician Galen (131- 201 CE) studied animals by cutting phrenic nerve (breathing)
Gladiators with injuries below neck are capable of breathing, opposite isn't true
• Julien Cesar Legallois was a vivisectionist (surgery on living animals). He decapitated
animal at different levels of brain stem, observed that headless body could breath and survive (5
hours). If it occurred further down brain stem, then respiration stopped completely
Respiration continued until he got to medulla
Identified brainstem, and medulla oblongata as essential for life
Theorized that breathing localized to medulla (untrue due to cardiovascular regulation)
• Dr. John Harlow studied his patient Pheneas Gage. Rod was driven through skull, frontal
lobes. Gage recovered with no focal neurological deficit besides blindness. Memory intact,
intellect declined, and personality changed (uncontrollable desire), loss of executive function
First observations how higher cognitive processes are localized to parts of the brain
Anatomy of a Neuron
• Neurons: Excitable cells that process, and transmit information via electrical and chemical
signals. Distinct specialized structures from other cells.
• Dendrite: Short, branched processes extending from soma and receive signals from other
neurons or chemical messengers. Transmit information as electrical information to soma.
• Soma: Cell body. Contains same organelles as other cells. Organelles restricted to this.
• Axons (Neurofibers): Single, long, slender projections that extend outward from soma and
conduct electrical signals away from cell body and toward target(s).
Nerves: Bundles of axons.
• Synaptic terminal: End of axon where electrical signal is converted to a chemical signal.
• Dendrites and soma are the receptive part of neuron
Glia
• Astrocytes: Largest, most prevalent type of glia cell. Found in CNS. Two roles.
Supportive role as a glial cell component in neural tissue
Indicates diseased tissue since reactive astrogliosis is a reliable, sensitive marker
• Elaborate processes extend from cell body to give star-like appearance
• Outnumbers neurons 5-fold
• Foot processes wrap around endothelial in capillary walls
Contributes to production and maintenance of blood-brain barrier
• Important in forming glial scar, very reactive to injury
• Markers have been used in research to determine diseased tissue
Glial Fibrillary Acidic Protein (GFAP) is a fairly accurate marker
Glutamate-Glutamine cycle
Types of Astrocytes
Myelin-forming glia
Microglia
• Older X-ray-based methods are used to image basic features of brain and cerebral vasculature
• Very little contrast, poor resolution, high dose of radiation
fMRI of a Patient’s Brain that Harbored a Right Frontal Lobe Glioma (Gray Area)
• fMRI provide live images the patient are asked to think about things, or ordered to do task
• Allows localization of tumor in relation to sensory cortex, and other structures
• Shows structure, and function of brain
Indirect “Images” of Functional Localization and Maps using Invasive and Noninvasive
Neurophysiological Approaches
• Intracranial surface electrodes: View brain by removing skull, but not penetrating the brain.
Constructed map of sensory, and motor cortex
Surface electrodes record electrical activity in brain
• Trans-cranial surface electrodes: View brain without removing skull.
Device generates magnetic field that delivers pulse to cerebral cortex (Mini-MRI)
Used to treat mood disorders
Single-unit electrophysiological recording in a monkey
• Single-unit recording: Electrode is inserted into cortex, doesn’t penetrate neuron but sits
outside it. Records neuronal activity. Records number of action potential discharges
• Skin is stimulated by probe. Firing rate of neuron in response to stimulus is recorded
• Maps cortical regions (sensory, memory)
• Dyes penetrate cell membranes and change color due to calcium or voltage change
• Displays how neuronal circuits work together by recording multiple neurons simultaneously
• in-vitro rather than in-vivo preparation
• Older dyes had slower response times which doesnt allow useful capture of activity
• Modern dyes have faster changing emission properties
Functional Relationships
• Environment —> Sensory system —> CNS —> Motor system —> Effectors
Anatomical Terminology
Neuroanatomical Directions
• Dorsal and ventral are altered in CNS since the brain makes a 90° turn
Dorsal is above midline of brain, ventral is below midline of brain
Organization of the Brain
• CNS originates from neural tube which grows, and develops. Timing is critical
• Prosencephalon (Forebrain): Develops into two key brain structures
Telencephalon: Becomes cerebrum (cerebral cortex), white matter, and basal nuclei
Overgrows diencephalon, reaches back to metencephalon.
Diencephalon: Become epithalamus, hypothalamus, and thalamus. (eye cups)
• Mesencephalon (Midbrain): Rostral part of brainstem. Contains visual, and auditory structures
• Rhombencephalon (Hindbrain): Divided into two parts.
Metencephalon: Pons (brainstem), and cerebellum
Myelencephalon: Medulla oblongata (brainstem)
• Cerebral cortex: Outer cerebrum layer composed of folded gray matter. Role in consciousness.
• Sulcus: Shallow invagination. Inside. Contains a bottom.
Sulci and gyri are unique as a face
• Gyrus: Exposed portion of tissue. Outside.
• Fissure: Deep invagination. Inside. Contains no bottom.
• Central sulcus: Divides frontal, and parietal lobe. Conserved location in humans.
• Lateral (Sylvian) fissure: Divides frontal, and temporal lobe. Conserved location in humans.
• Parieto-occipital sulcus: Divides parietal, and occipital lobe. Conserved location in humans.
• Cingulate gyrus: Fold of tissue above corpus callosum.
• Cingulate sulcus: Separates the frontal, and parietal lobes from the cingulate gyrus.
• Pre-occipital notch: Located after occipital lobe, and before temporal lobe.
• Hippocampus: Memory center. Controls learning, long-term memories, and remote memories.
• Brainstem: Consists of medulla, pons, and midbrain.
• Amygdala: Combat center. Involved in processing memory, decision-making and emotional
response (fear, aggression)
• Striatum: Coordinates aspects of cognition including motor, action planning, decision-
making, motivation, reinforcement, and reward perception. Component of basal ganglia.
Planes of Section
Oblique Cut
• Encephalon: Brain.
• Prosencephalon: Forebrain. (cerebrum)
Telencephalon: Endbrain.
Contains cerebral hemispheres
Cerebral cortex, white matter, basal ganglia, and lateral ventricles
Diencephalon: Inter-brain.
Contains thalamus, hypothalamus, and third ventricle
• Rhombencephalon: Hindbrain. (brainstem)
Mesencephalon: Midbrain.
Contains substantia Nigra, cerebral peduncle, and cerebral aqueduct
Metencephalon: After-brain.
Contains pons, cerebellum, and fourth ventricle.
Myelencephalon: Medulla.
Contains medulla oblongata, and fourth ventricle
• Brain is divided into 2 major divisions, and 5 sub-divisions
Each subdivision contains parts of ventricular system where CSF is created and flows
1. Frontal lobe: Controls all voluntary movement, perception, problem-solving, and reasoning.
Prefrontal cortex: Controls executive function, impulse control, and long-term planning.
Motor cortex: Controls skeletal muscles. Located on precentral gyrus.
Broca’s area: Controls speech production.
2. Parietal lobe: Controls touch, pressure, temperature, pain, spatial processing, and orientation.
Somatosensory cortex: Somatosensory processing. Located on postcentral gyrus.
3. Occipital lobe: Visual centers of brain. Sits on top of cerebellum.
4. Temporal lobe: Controls sound processing, speech perception, short-term memory, emotion.
5. Limbic lobe: C-shaped region that crosses brain hemispheres within the cortex, including
portions of the temporal, parietal, and frontal lobes.
• Grey matter: Composed of somas, dendrites (input fibers), and axons (output fibers)
Not confined to the cerebral cortex, found in deeper structures as well
Stained red with Eosin dye
• White matter: Composed of axons coated in myelin.
Stained blue with Luxol Fast Blue
• Sometimes grey and white matter mix together like in the Reticular Formation
• Gray matter is present on outside, white matter is internal
• Fixed: Submerged in fixed formaldehyde
Using compound that labels myelin proteins
White matter is dark blue, gray matter is pale pink
Cells of the Cerebral Cortex
Pyramidal Neuron
• Dendrites are not smooth, they have mini processes coming off called dendritic spines.
Each one is a synapse
• They are dynamic, and change with activity
• This is where learning takes place since they change shape due to being used
Astrocytes
• Astrocytes are a type of glial (support) cell that has many functions.
• They create circuit boxes around synapses to allow and control communication
• They coat both neurons and capillaries, thus influencing brain metabolism.
• They serve as “guide-wires” during the development of the cerebral cortex and other brain
regions, guiding cells to their proper place.
• They also react to damage caused by neurological diseases, stroke, and trauma.
Reactive Astrocytes
• Reactive astrocytes lack star-like appearance. Cell body and processes hypertrophy, forming
scar-like structures
• Takes up debris, produces signals to attract microglia. They grow into damaged site to form
a scaffold to seal-off injury. Adheres tissue, provides barrier to prevent axons from growing
Microglia
• Microglia: Brain’s surveillance cells. Immune system. Monitors disease, and tissue damage.
• When they detect a pathological change, they multiply, migrate to the diseased or damaged
site, and engulf and digest the pathogens/debris
• Brain’s blood is supplied from four arteries: 2 internal carotid arteries and 2 vertebral arteries
• Upon reaching forebrain, the internal Carotid gives the Anterior and Middle cerebral arteries
• Two vertebral arteries reach the ventral surface of the brainstem and give off the first pair of
Cerebellar arteries before coming together to form the Basilar artery
• Basilar artery gives rise to two pairs of Cerebellar arteries and Posterior Cerebral arteries
• Basilar artery: Arise from vertebral arteries. Runs along midline of pons (major supplier).
• Vertebral artery: Arise from the subclavian arteries.
• Common carotid artery: Arise from aorta. Supplies blood to head, and neck.
• Internal carotid artery: Arise from common carotid artery. Supplies blood to brain, and eyes.
• External carotid artery: Arise from common carotid artery. Supplies blood to face, scalp, skull.
• Subclavian artery: Arise from the aorta.
• Flow of blood: Vertebral artery —> Basilar artery —> Posterior communicating artery —>
Middle cerebral artery —> Anterior communicating artery —> Anterior cerebral arteries
• Posterior inferior cerebellar arteries: Branch off vertebral artery. Supplies blood to cerebellum.
• Anterior inferior cerebellar arteries: Branch off basilar artery. Supplies blood to cerebellum.
• Pontine arteries: Branch off basilar artery. Supplies blood to the pons.
• Superior cerebellar arteries: Branch off basilar artery. Supplies blood to cerebellum.
• Posterior cerebral arteries: Branch off basilar artery. Supplies blood to occipital lobe.
• Anterior choroidal artery: Branch off internal carotid artery. Supplies blood to lobes.
• Ophthalmic artery: Branch off internal carotid artery. Supplies blood to eyes.
• Middle cerebral arteries: Branch off internal carotid artery. Supplied blood to lobes.
• Circle of Willis: Three cerebral, two communicating, and one carotid artery.
Clinical importance in headaches (applied pressure to optic nerves, hypothalamus)
Brain’s Ventricular System
• Brain ventricles: Creates CSF, and flows through ventricles into spinal cord.
• Flow of CSF: Lateral ventricle —> Third ventricle —> Cerebral aqueduct —> Fourth ventricle
• Most rostral part are lateral ventricles
• Forebrain grows forward, then back (Lateral ventricle follows that C shape)
• Anterior (frontal) horn of lateral ventricle: Connects to 3rd ventricle. Impinges into frontal lobe
• Inferior (temporal) horn of lateral ventricle: Impinges into temporal lobe.
• Posterior (occipital) horn of lateral ventricle: Impinges into occipital lobe.
• All ventricles are continuous
• No ventricles in the occipital lobe
• Ventricle system: Communicating network of cavities filled with cerebrospinal fluid (CSF) and
located within the brain parenchyma. Four main regions.
1. Lateral ventricle: Two. Largest. One in each cerebral hemisphere.
Under corpus callosum and above fornix
Connected to the third ventricle by interventricular foramen (foramen of Monro)
2. Third ventricle: Separates the two thalami and hypothalami.
Located in the diencephalon, midline.
3. Cerebral aqueduct: Connects the third, and fourth ventricles.
Located in the mid-brain, separated by colliculus and mesencephalon
4. Fourth ventricle: Continuous with central spinal canal, and subarachnoid layer.
Located between cerebellum and lower brainstem
Choroid Plexus
• Cells of the choroid plexus receive blood from capillaries, transform blood into CSF, and
release CSF into the ventricles.
• Convective flow of CSF washes brain which removes debris, and toxins
Accelerates during certain sleep stages. Sleep deprivation reduces convective flow
Brain’s Protective Coverings: The Meninges
Relationship of the spinal cord and spinal nerves in the vertebral column
• Myelin is flipped compared to cerebrum (Gray matter on inside, white matter on outside)
• Gray matter is arranged in layers, and forms butterfly structure
• Sensory information is collected by sensory ganglia. Their dendrites extend to periphery,
axons extend to gray matter of dorsal horn.
• In myotatic reflex, tensor and flexor cell bodies reside in ventral, interneuron in dorsal.
Interneurons housed entirely in gray matter of spinal cord
• Sensory and motor information travel within the same nerve
• Gray matter in spinal cord arranged in specific layers (Rexed Laminae), or subdivisions
• Dorsal horn
Lamina II: Substantia gelatinosa. Modulates pain pathways.
• Ventral horn
Lamina IX: Motor neuron columns that govern limb musculature.
Learning Objectives (Week 1)
1. Describe the historical evolution of the essential ideas about the nervous system in
terms of neurons as excitable cells.
Cell theory proposed by Schwann and Schleiden. Joseph von Gerlach devised protoplasmic
network (reticular theory).
Neuron theory: CNS made up of individual, discrete parts. Neuronists, or reductionists. Santiago
Ramón y Cajal.
Reticular theory: CNS acts as a single unit, not discrete parts. Joseph von Gerlach.
Protoplasmic network. Holistic interpretation.
Localization of function: Brain has discrete regions that serve specific functions
Dr. Broca: Discovered Broca’s area by studying expressive aphasia, and lesions
Galen: Studied animals by cutting phrenic nerve (breathing), noticed decapitated gladiators
Legallois: Vivisectionist. Identified brainstem, and medulla oblongata as essential for life.
Dr. Harlow: Showed that higher cognitive processes were localized. Pheneas Gage impaled.
4. Describe techniques used to study nervous system and brain organization/
connectivity
X-ray: Photographs basic features of brain and cerebral vasculature. Very little contrast, poor
resolution, high dose of radiation.
b. CAT scan
CAT scan: X-ray source and detector moved around subject. Non-invasive imaging that makes
horizontal slices. Greater resolution of brain structures.
MRI: Safe, non-invasive technique. Provides greater resolution, and clarity. Magnetic field aligns
water molecules, and radio-waves produce resonance of water molecules
fMRI: Provides live images while patients think about something. Allows localization of tumor in
relation to other structures. Shows structure, and function of brain
Intracranial surface electrodes: Skull removed, electrodes placed on brain which records
electrical activity. Constructs map of sensory, and motor cortex.
Transcranial magnetic stimulation: Device placed on outside of skull that generates magnetic
field, and delivers pulses to brain. Treats mood disorders.
Single unit recording: Electrode inserted into cortex, but doesn’t penetrate neurons. Records
neuronal activity. Skin stimulated by probe, and neuronal firing rate is recorded.
Fluorescent dye imaging: Dyes penetrate cell membranes and change color due to calcium or
voltage. Records multiple neurons simultaneously to show how neuronal circuits work together.
Cerebral cortex: Outer layer of cerebrum which is folded gray matter. Role in consciousness.
Sulcus: Shallow invagination. Inside. Contains a bottom.
Gyrus: Exposed portion of tissue. Outside.
Fissure: Deep invagination. Inside. Contains no bottom.
Central sulcus: Divides frontal, and parietal lobe. Conserved location in humans.
Parieto-occipital sulcus: Divides parietal and occipital lobe.
Lateral (Sylvian) fissure: Divides frontal, and temporal lobe. Conserved location in humans.
Cingulate gyrus: Fold of tissue above corpus callosum.
Cingulate sulcus: Separates the frontal, and parietal lobes from the cingulate gyrus.
Pre-occipital notch: Located after occipital lobe, and before temporal lobe.
Longitudinal fissure: Separates two cerebral hemispheres
Precentral gyrus: Front-half of central gyrus that carries out motor cortex.
Postcentral gyrus: Back-half of central gyrus that carries out sensory cortex.
Cerebellum: Top of hindbrain. Integrating center. Role in posture, coordination, and balance.
Pons: Above medulla. Relay between the cortex and the medulla. Controls balance.
Medulla oblongata: Bottom of hindbrain responsible for regulating vital functions.
Mammillary body: Caudal end of hypothalamus.
Optic chiasm: Rostral end of hypothalamus. Visual tracts cross.
Corpus callosum: White matter that connects both sides of brain.
c. Distinguish gray matter from white matter
Grey matter: Composed of somas, dendrites (input fibers), and axons (output fibers)
White matter: Composed of axons coated in myelin.
Glia: Glue for nervous system. Maintains extracellular ions, modulates signal conduction,
regulates neurotransmitter uptake, scaffold for neural development, interface for immune
system, convective flow, and involved in neural injury
Astrocytes: Most prevalent type of glia cell found in CNS. Star-like. Facilitates Glutamate-
Glutamine cycle, creates circuit boxes around synapses, coat both neurons and capillaries,
“guide-wires” during development, and reacts to damage to form scars.
Dura Mater: Outermost layer. Tough mother, very durable. Contains venous.
Arachnoid layer: Middle layer. Spider, thin, and web-like appearance.
Sub-arachnoid space: Contains CSF and blood vessels
Pia Mater: Inner-most layer. Tender mother, very delicate. Follows contours of brain closely.
g. Identify the following structures of the human brain on images
i. Forebrain
ii. Midbrain
iii. Hindbrain
iv. Telencephalon
1. The five cortical lobes
2. Major sulci and gyri
v. Diencephalon
vi. Mesencephalon
vii. Metencephalon
viii. Myelencephalon
ix. Brainstem
x. Cerebral hemispheres
xi. Corpus callosum
xii. Cerebellum
xiii. Thalamus
xiv. Hypothalamus
xv. Optic tract
xvi. Optic chiasm
xvii. Mammillary body
xviii. Superior colliculus
xix. Inferior colliculus
xx. Pons
xxi. Medulla oblongata
xxii. Ventricular system
Weeks 2-4: Neural Signaling
• All living cells maintain a potential difference across their cell membranes (Vm)
• Inside of the cell is usually negative relative to the outside
• In nerve cells the value of the resting membrane potential is between -40 and -90 mV
• Where does this potential difference across the plasma membrane come from?
• Ionic pathways for Na+, K+, and Cl- are required to establish resting membrane potential (Vm)
• Chemical gradient: Difference in concentration across a membrane.
• Electrical gradient: Difference in charge across a membrane. Initially does not exist, but
established as soon as K+ begins to move from internal into external compartments.
• Active transporters: Actively moves ions against concentration gradient. Maintains Vm.
• Ion channels: Allows ions to diffuse down concentration gradient. Selective permeability.
Establishes membrane potential
Electrochemical equilibrium
• Membrane only permeable to K+. No net movement during 1 mM inside, and 1 mM outside
• Increasing the KCl concentration of the inside compartment to 10 mM initially causes a small
movement of K+ into outside compartment (initial conditions) until electromotive force acting on
K+ balances concentration gradient, and there is no further net movement of K+ (equilibrium)
Simple Derivation of the Nernst Equilibrium Potential
• Equation calculates Veq based on nature of ion (valence) and ion chemical gradient
• Free energy of chemical gradient (ΔGchemical) = RT * ln( [X]o / [X]i )
R = Gas constant: 8.314 J per kelvin per mol
T = Absolute temperature in Kelvin (K = C + 273.15)
• Free energy of electrical gradient (ΔGelectrical) = zFVeq
Free energy of electrical gradient
z = Valence of ion, unit-less
F = Faradays constant: 96,485 Coulombs per mol
V = Voltage
• Chemical and electrical are equal and opposite at equilibrium, ΔGelectrical = ΔGchemical
zFV = RT * ln( [X]o / [X]i )
• Final Nernst Equation : V = RT/zF * ln( [X]o / [X]i )
• This equation calculates potential established across membrane based on valence and
chemical gradient, if only one type of ion channel is present. This potential known as Nernst
potential or Nernst equilibrium potential. Only one ion present
• Nernst (equilibrium) potential (Veq): Vm at which ionic species is at equilibrium
• Common to use ion symbol as subscript to denote Veq for that ion (VK, VNa, VCl)
• If only one ion present, and channels are present and open, then Veq = Vm
• ENa = +64 mV, EK = -90, ECl = -87, ECa = +160
• If two or more ions contribute to Vm, then Nernst no longer yields Vm, must use GHK
• The Nernst equation gives a formula that relates the numerical values of the concentration
gradient to the electrical gradient that balances it.
Model Cell
Nernst equilibrium
• Defines the electrical potential across a membrane that will balance a particular concentration
gradient of an ion to which the membrane is permeable
• Synonymous with equilibrium potential and also called the reversal potential
• Depends upon the concentration gradient and the ion valency; does NOT depend on the
degree of permeability of the membrane to the ion
• Relatively few ions have to move to set up the potential; concentration gradients are not
significantly disturbed in achieving an equilibrium
• If a membrane is permeable to only ONE type of ion, then the membrane potential will
automatically move to the equilibrium potential of that ion.
• Assumes permeability therefore it left out of equation. Derived from chemical and electrical
• Always check units, convert to mM
Understanding Leak channels
• Leak channels: Ion pathways across plasma membrane which establish Vm. Controls Vrest
• Very little alters permeability characteristics of leak channels
• They are not gated, therefore it is constant background permeability
• At rest (cell is unperturbed), permeability of membrane goes from K+ > Na+ > Cl-
• Not known if separate leak channels for each ions, or single leak channel with different
permeability properties (K+ passes relative to [Na+] and [Cl-])
• Electrochemical gradients contribute to Vrest
• Na+-K+ ATPase is important in maintaining chemical gradients
• Each ion has its own Nernst potential which is different from all other ions
• The membrane potential is a “compromise” between the various equilibrium potentials,
each weighted by the membrane's permeability and absolute concentration of the ion.
• At rest, Vm is dominated by potassium because of large PK
• There will be a flux of ions across the membrane of any ion for which the membrane potential
(Vm) is not at the equilibrium potential (Veq)
• If Vm is stable, then the total net flux of ions across the membrane is zero
Driving Force
• When ion is not at equilibrium, an electrochemical driving force (VDF) acts on the ion, causing
the ion to flux across the plasma membrane according to its own electrochemical gradient.
• Equation: VDF = Vm - Veq
Veq calculated from Nernst equation if concentration and temperature provided
Vm calculated by Goldman equation if all variable information provided
Example 1: DFK
Example 2: DFCl
• Given: Vm = Vrest = -68 mV, VK = -97 mV, VNa = +61 mV, VCl = -64 mV
• DFK = Vm - VK = -68 - (-97) = +29mV (K+ efflux)
• DFNa = Vm - VNa = -68 - (+61) = -129mV (Na+ influx)
• DFCl = Vm - VCl = -68 - (-64) = -4 mV (Cl- efflux)
• Asymmetric ion distribution and selective ion channels are needed to establish Vm
• In most cells, Vrest determined mainly by K+ and Na+, may have Cl- contribution
• Since in most cells, Vm =/= Veq for any ions, at rest each ion moves down its electrochemical
gradient due to DF acting on ion (replenished by active transport)
• Vm = Vrest = -68 mV, VK = -97 mV, VNa = +61 mV, VCl = -64 mV
• DFK = Vm - VK = -68 - (-97) = +29mV (K+ efflux)
• DFNa = Vm - VNa = -68 - (+61) = -129mV (Na+ influx)
• DFCl = Vm - VCl = -68 - (-64) = -4 mV (Cl- efflux)
• Asymmetric ion distribution and selective ion channels are needed to establish Vm
• In most cells, Vrest determined mainly by K+ and Na+, may have Cl- contribution
• Since in most cells, Vm =/= Veq for any ions, at rest each ion moves down its electrochemical
gradient due to DF acting on ion (replenished by active transport)
Na+-K+ ATPase Inhibition
• Under resting physiological conditions, constant [K+] efflux and [Na+] influx, in regards to cell
This dissipates gradients if ionic concentrations are not maintained
• Primary active transporter pumps K+ into and Na+ out of cell (Na+-K+ ATPase) which
counteracts constant movements of these two ions due to electrochemical gradients
• Moves 2 K+ into cell and 3 Na+ out of cell, creating electrogenic transport process every
transport cycle, 1 net positive charge is transmitted from inside to outside
Magnitude varies from cell to cell, depends on other ionic pathways
• Na -K+ ATPase activity is inhibited by chemical agent (Ouabain, Vanadate), Vm is initially
+
slightly depolarized (> 5 mV) due to lack of ATP hydrolysis and transport cycle
Does not immediately abolish membrane potential
• 1. Vrest = -68 mV —> 2. Ouabain applied, modest depolarization of cell, Vm = -63 mV —>
3. Cell not able to pump for long period of time, Vm becomes less negative until it completely
disappears, Cell is dead. Constant Na+ influx and K+ efflux abolishes their chemical gradients.
• In absence of inhibitors, Veq is close to 0, leading Vm to 0. To maintain Vm, cells have to expend
energy to maintain proper intracellular Na+ and K+ concentrations.
• Na+-K+ ATPase is not responsible for generation of Vm, it is responsible for maintaining it, by
maintaining normal intracellular K+ and Na+ concentrations
Na-K ATPase is an electrogenic pump (-)
• Cells expend energy to maintain [Na+]i and [K+]i, through activity of Na+-K+ ATPase in order
to maintain Vrest. Vm = Vrest is essential for many physiological processes
• Action potential (electrical impulse) is responsible for nervous system communication.
Involves rapid reversal of Vm, intracellular is positive with respect to outside
Vm has to depolarize from -70 mV to -50 mV
At Vthreshold, voltage-gated sodium and potassium channels can be activated
Normal action potential generation therefore depends on normal value of Vrest
• Voltage-gated sodium channels of excitable cells enter inactivated, non-conducting state after
opening. Can not re-enter open state without recovering to closed state.
Recovery is voltage-dependent, Occurs when Vm is more negative than Vrest
Return of Vm to Vrest is critical for allowing channels to continue functioning
• If condition causes change (increased [K+]o), voltage-gated sodium channels can not recover
after being activated and subsequently inactivated from action potential
Neurons and cardiac myocytes can no longer function. Catastrophic for organism
Na+-Glucose Cotransporter
• Na+-Glucose cotransport is electrogenic, and Vm sensitive since 2 Na+ export each cycle
• At Veq, there is no net transport of Na+ and Glucose, Vm = Vrev (reversal potential)
Vm is rarely equal to Vrev, therefore transporters are not at equilibrium at Vrest
No net transport if Vrest = Vrev
• Reversal potential: Vrev = RT/2zNaF * ln ( ([Na+]o2 * [Glucose]o) / ([Na+]i2 * [Glucose]i) )
• Driving force acting on transport: DFT = Vm - Vrev
• If Vm is more negative than Vrev, transporter cotransports Na+ and glucose into the cell.
Referred to as Sodium-coupled transporter forward mode of transport
Under normal physiological concentrations, SGLT works in forward mode.
• If Vm is more positive than Vrev, transporter cotransports Na+ and glucose out of the cell.
Referred to as Sodium-coupled transporter reverse mode of transport
• Direction of transport changes forward to reverse depends on Vm (DF acting on transporter)
• Since Vm is rarely equal to Vrev, and [K+]o and [K+]i are regulated within narrow limits, Glucose
accumulates intracellularly. SGLT works in forward mode to achieve Vrev = Vm
• Concentrative transporter capacity: [Glucose]i / [Glucose]o = ( [Na+]o2 / [Na+]i2 ) * e^(-2VmF/RT)
SGLT is described as ratio of [Glucose]i to [Glucose]o
Vm influences concentrative capacity of the Na+-Glucose transporter
As Vm becomes more negative, concentrative capacity increases
If Vm not maintained at large negative values, CC is reduced substantially
• Concentrative capacity provides measure of how well transporter cumulates substrate in cell
against chemical gradient, given that [K+] are fixed due to cellular homeostatic processes
• Vm provides significant free energy for glucose accumulation inside epithelial cells of small
intestine and kidney proximal tubules, complete removal in lumen before ultra filtrate moves on
• For cell with Vrest = -50 mV: [Glucose]i / [Glucose]o ~4000
SGLT can concentrate glucose ~4000 times higher inside cell than outside
If Vm collapses then glucose can not be concentrated in the cell
2.2 - Passive Electrical Properties of the Neuron
Overview
• Excitable cells (neurons, muscles) generate electrical signals conducted over long distances
• Nernst and GHK equations only apply to steady state conditions (constant voltage)
• Stable equilibrium state: Intracellular and extracellular solutions must be electrically neutral,
in osmotic balance, and have no net movement of ions.
• Non-steady state conditions include action potentials, synaptic potentials, and sensory
generator potentials
Questions to consider
• They provide a return pathway to complete electrical circuit when active currents flow in/out
Determines time course and amplitude of potential generated by synaptic current
• They determine whether a synaptic potential generated at one location of a neuron (e.g. a
dendrite) will result in a supra-threshold depolarization
• They influence the speed at which an action potential is conducted down the axon
Input resistance
• Internal & external ion solutions act as two conductors, while phospholipid bilayer acts as
a capacitor since it is a good, thin insulating layer. High capacitance, separation of charge.
Membranes as Resistors
• Negative charges accumulate on the inside, positive charges accumulate on the outside
• Capacitance of the membrane has the effect of reducing rate at which the membrane
potential changes in response to current
• If the membrane had only resistive properties, a step pulse of outward current passed
across it would change the membrane potential instantaneously
• If the membrane had only capacitive properties, the membrane potential would change
linearly with time in response to the same step pulse of current.
• Because the membrane has both capacitive and resistive properties in parallel, the actual
change in membrane potential combines features of the two pure responses
• Initial slope of the relation between Vm and time reflects a purely capacitive element,
whereas the final slope and amplitude reflect a purely resistive element
• Since the resistance and capacitance of the membrane are in parallel, the voltage across each
element must always be the same and equal to the membrane potential
• Capacitance (C): C = Q / V
Q = Charge (Units: Coulombs)
V = Voltage
• Capacitance (C): C = 𝜀0 * (A/d)
A = Area of plate
d = Distance of plate separation
↑ A = ↑C
d=↓C
𝜀 = Permittivity or dielectric constant
• Smaller neuron with smaller area has a shorter time to change Vm which allows faster
conduction velocity
Capacitance in Electrical Circuit
• V = Q / C —> ΔV = ΔQ / C
• Ic = ΔQ / Δt —> ΔQ = Ic * Δt
Ic = Current flow across capacitor
• ΔV = Ic * Δt / C
• Magnitude of voltage change across capacitor in response to a current depends on the
duration of current, as time is required to deposit and remove charge on plates of the capacitor
• Capacitance is directly proportional to area of the capacitor plates. Larger area will store more
charge for a given potential difference
• Because capacitance increases with cell size; More charge, and therefore current, is required
to produce the same change in membrane potential in a larger neuron than in a smaller one
• Im = Ii + Ic
Ii = Ionic (resistor) membrane current
Ic = Capacitive membrane current
Im = Total membrane current
• Length constant (λ): Distance at which ΔVm decayed 37% compared to point of injection.
Conductance-based membrane model
• V = I/R = gV
• Capacitor: Phospholipid bilayer
• Resistor: Ionic permeabilities (channels)
• Battery: Electrochemical driving forces
• Kirchhoff’s law: Total current flowing across the cell membrane is the sum of the capacitate
current and the ionic currents.
Equation: Im = Ii + Ic
3.1 - Action Potential
• The Hodgkin-Huxley (HH; Hodgkin & Huxley, 1952) model for the generation of nerve action
potentials is one of most successful models of a complex biological process ever formulated
• Basic concepts expressed in the model have proved a valid approach to the study of
bioelectrical activity from the most primitive single-celled organisms such as Paramecium, right
through to the neurons within our own brains.
• Squid axon is very large (1 mm) which is why it was used as a first model
• Insertion of glass electrodes down center of axon, recorded electrical events within axon
• Demonstrated that electrical events were extremely fast by comparing to sin wave frequency
• Changing ion concentrations of solution within squid changed action potential events
• In order for an ion channel to open, all gates within that channel must open simultaneously
• Individual gates open and close randomly and VERY rapidly
• Open probability: Probability of a gate being open. Dependent on voltage across membrane.
• Activation variable: Probability of a gate being open at any point in time for that gate.
• Gates carry charges themselves and their position in the membrane is affected by the electric
potential across the membrane
1. One internal electrode measures Vm, and connected to voltage clamp amplifier.
2. Voltage clamp amplifier compares Vm to desired (command) potential.
3. Clamp amplifier injects current through a second electrode when Vm differs from command
potential. Feedback causes Vm to become command potential.
4. Current flowing back into axon, and thus across its membrane, is measured
• Two electrodes inside squid axon (only one needed for small neuron)
• Reference electrode hooked up to ground, used as a reference
Current flow across a squid axon membrane during a voltage clamp experiment
• IV relationship
• Late portion is the outward current. Due to potassium
Continuously gets larger
• Early portion is the inward current. Due to sodium
Gets larger, then smaller, switches direction, then gets bigger
Current reverses flow at equilibrium potential for sodium
Dependence of the early current on sodium
• Inward current becomes outward because the driving force on sodium changes (no sodium
outside, only inside, driving force is now outward)
• Driving force determines direction of ions
• Experiment shows that there is another ion participating that is unaffected by sodium
• Depolarizing membrane potential produces early Na+ influx followed by delayed K+ efflux
• Differences in time course suggest there are two different mechanisms activated by
depolarization
Both channels activated by voltage, but different mechanisms of activation
A mathematical description of permeability changes across the plasma membrane
Membrane conductance changes underlying the action potential are time- and voltage-
dependent
Hodgkin and Huxley Voltage Clamp Studies Key points:
Mathematical reconstruction of the action potential
• Resting: m-gate is closed (open probability low), h-gate is open (open probability high)
• Activated: m-gate is open (open probability high), h-gate is open (open probability decreasing)
• Inactivated: m-gate is open (open probability high), h-gate is closed (open probability low)
• h-gate moving slowly gives sodium time to flux across the membrane
• To get back to resting state, membrane requires repolarization
Voltage gated sodium channels: activation, inactivation, and recovery from inactivation
• Closed channel —> Open channel —> Inactive —> Closed channel
• n-gates: Activation gates. 4 of them. Open probability increases with depolarization of Vm.
Much slower kinetics, lacks inactivation gate
• Called delayed rectifier
Threshold
• Membrane depolarization —> pNa increase —> Na+ influx —> Membrane depolarization
• Positive feedback loop
• Increasing open probability of m-gates
• Cycle stops due to potassium, and inactivation of sodium channels
• Fast positive cycle: Depolarize Vm —> Open Na+ channels —> Increase Na+ current —>
Depolarization
• Slow negative cycle: Depolarize Vm —> Open K+ channels —> Increase K+ current —>
Hyperpolarization
Refractory period
• Refractory period: Time where cell can not activate another action potential.
• Absolute refractory period: Voltage-gated sodium channels unavailable to open again.
• Relative refractory period: Some voltage-gated sodium channels available to open, but stimuli
needs to be larger than initial threshold stimuli.
3.2 - Conduction & Propagation
Why does a voltage signal applied to one end of a nerve fiber fail to spread?
• Membrane resistance (Rm): Resistance across membrane wall, describes extent of current
leaking through membrane. Connected in parallel with capacitor.
• Longitudinal resistance (Ri): Resistance within axon cytoplasm. Connected in series.
• Extracellular compartment is so large that it is assumed to have zero resistance
• Changing parameters of fiber affects resistances
• Example: Longer processes, Ri & Rm decrease due to added leakiness, Cm increases due to
greater area
The length constant (λ) depends on both Rm and Ri
• λ = (Rm/Ri)1/2
• Distance over which the potential change spreads increases with increasing Rm
Prevents leakiness to allow current flow
• Distance over which the potential change spreads decreases with increasing Ri
Difficult for charges to travel along axoplasm
• Squid axon:
Rm = 2000 Ωcm2, Axon diameter = 1 mm, Ri = 30 Ωcm, λ = 13 mm
• Frog axon:
Rm = 2000 Ωcm2, Axon diameter = 50 μm, Ri = higher, λ = 1.4 mm (lower)
• Mammalian axon:
Rm = 2000 Ωcm2, Axon diameter = 1 μm, Ri = MUCH higher, λ = 0.3 mm (MUCH lower)
• Diameter is the only parameter that is changing. λ decreases, Ri increases
• Rinput and λ determine the size of the signal generated in a nerve process (V = IR)
• Rinput and λ determine how far a signal will spread
• Rinput and λ depend on fiber size (determines resistance to flow along core)
• Rinput and λ depend on resistive properties of the cytoplasm and plasma membrane
• Example: Small dendrite will have larger Rinput, larger current, larger voltage change
Larger dendrite has a longer length constant, smaller voltage change
• Action potentials are able to propagate without loss in size. Same amplitude and size,
propagate without degradation
• Action potentials don’t depend on passive properties on membrane. Propagation depends on
active processes
Voltage dependent feedback cycles
• Same feedback cycles responsible for generating the action potential are responsible to
propagation of electrical signals down the axon.
• Fast positive cycle: Open Na+ channels —> Increase Na+ current —> Depolarization
• Slow negative cycle: Open K+ channels —> Increase K+ current —> Hyperpolarization
• Distance current flows depends on: 1. Duration of action potential, 2. Conduction velocity
• Depolarization phase due to Na+ movement, repolarization phase due to K+ movement
K channels stay open until depolarization is over, takes longer
• Wave of depolarization followed by wave of hyperpolarization
• Ion flux activates K+ channels along wave of depolarization. Inactivation of Na+ channels, and
activation of K+ channels creates a trailing dip in Vm
• Distance depends on duration of action potential, and conduction velocity of fiber
• Action potentials only travel one direction due to refractory period of cell
• AP’s last 2 ms, conduction rate is 10mm/ms, AP spreads about 20mm. Passive travels 2 mm.
• AP traveling longer distance requires greater conduction velocity
Requires different size or different resistance or different capacitance
Capacitance influences time course of current change
• Depolarization opens voltage-gated Na+ channels and leads to the delayed opening of
voltage-gated K+ channels as well as Na+ channel inactivation
• As AP travels down axon, some aspects are passive
Active process is depolarization and repolarization
Passive process is spread of current to adjacent portions of process
• Current spreads via diffusion
Conduction Velocity
Myelinated axons
• Myelin occurs on axons, not dendrites. Larger axons tend to be myelinated over smaller ones
• Lamellae: Myelin wrappings. Two layers of plasma membrane with no organelles.
Number of lamellae ranges from 10-20 to a maximum of ~1605
• Wrapping of 160 lamellae = 320 membranes in series between axon membrane and ECF
• Effective membrane resistance (Rm) is increased by a factor of 320 (Less current leaks out)
• Membrane capacitance (Cm) is reduced by a factor of 320 (Increased thickness of membrane)
Reduced capacitance reduces number of charges that need to be discharged
Speeds up rate of signal propagation down axon
• Nodes of Ranvier: Gaps in the myelin sheath where axonal membrane is exposed to ECF.
• Inter-node: Distance between nodes of Ranvier, ranges 200 μm to 2 mm.
Myelin
• Myelin impacts both passive and active properties of the axon membrane:
• Passive: Increases Rm, decreases Cm
• Active: Sculpts membrane organization of ion channels, and restriction of AP regeneration
Consequences of Myelination on Conduction Properties of the Axon
Demyelination and the fate of the axon
• Transient event allows other oligodendrocytes to remyelinate the axon, although new myelin
is thinner, and shorter than original. Possible recovery from disease.
• Long-term event leads to neuron degeneration. Slowed conduction velocity, reduction in
firing frequency, and eventual loss of neurons themselves
• Multiple sclerosis (auto-immune disease) causes demyelination of axons within CNS due to
immune system attacking myelin. Conduction velocity in non-myelinated axons is reduced.
4.1 - Synaptic Physiology
Electrical Synapses
• Electrical synapse: Gap junctions (closer than normal) in neurons that allow communication.
Links cytosol of two neighboring cells together which creates large conductance
Electrically, and chemically coupled together
• Connexons: Composed of 6 connexins: 1 set of 6 pre-synaptically, 1 set of 6 post-synaptically
Each cell contributes 6 connexins (half of full connexon), 21 connexin genes (GJA-GJE)
Connexons form plaques on neurons with multiple gap junctions present
• Connexin: 4 membrane spanning domains
• Presynaptic terminal: End of neuron where action potential was originated
• Postsynaptic terminal: End of neuron that coupled potential from another neuron
• Current flows passively (diffusion) through connexons
• Rapid transmission. Faster than chemical synapses, important in escape reflexes
Minimal delay (0.1ms) between presynaptic and postsynaptic neurons
• Bi-directional communication, presynaptic can turn into postsynaptic
• Cell synchronization. Syncing excitability in one cell to another (Smooth, cardiac muscle)
• Transfer of key molecules directly, passage of small molecules and ions
Second messengers (ATP, cAMP, Ca2+) can all diffuse through pores
• Transmission less likely to fail to toxins or synaptic depression (reliable)
• Capable of modulation by chemical messengers. Not just conduits of current
• Weaker than chemical synapses, therefore not prevalent in CNS
Chemical Synapses
• Chemical synapse: Secretion of neurotransmitter via synaptic vesicles. Most common type.
Different structure and function than electrical synapse
• Electrical signal in presynaptic cell communicated to postsynaptic cell via chemical
messenger (neurotransmitters). NT released from presynaptic terminal, diffused across
synaptic cleft, interacts with receptors on postsynaptic membrane.
• Presynaptic terminal: End of neuron where neurotransmitter is being released. Synaptic
vesicles clustered around region of exocytosis, active zone or site of release
Active zone: Site where vesicles fuse with pre-synaptic membrane
• Postsynaptic density: Opposite of active zone. Intracellular elements responsible for
maintaining postsynaptic receptors at proper locations in postsynaptic cell.
• Synaptic cleft: Separation between presynaptic and postsynaptic membrane (20-30 nm)
• Relies on diffusion across gap so they can interact with receptors anchored on postsynaptic
Since it is a small distance, diffusion is rapid
• Significant delay in signal (1 ms) but far more flexible than electrical synapse
Possible to use multiple chemical messengers, and multiple receptors
Far greater power in chemical than electrical
Synaptic Vesicles
• Active zone: Portion of the presynaptic membrane opposite the postsynaptic density across
the synaptic cleft. Site of synaptic vesicle docking, and fusion which leads to neurotransmitter
release. Three principle functions which are crucial for synaptic transmission.
1. Physically link synaptic vesicles, voltage-gated calcium channels, and other presynaptic
components into a single complex, thereby ensuring efficient co-localization of the elements
that are required for fast synaptic transmission.
2. They organize autocrine communication. Controls presynaptic receptors to allow
presynaptic modulation of neurotransmitter release. (self-regulation)
3. They mediate various forms of short- and long-term presynaptic plasticity.
Learning occurs with strengthen some synapses, and weakening other
Populations of Presynaptic Vesicles and the Kinetics of Exocytosis
• Three presynaptic vesicle pools. They vary in size and kinetics of exocytosis
• Reserve pool: (80-90%), Deposit of synaptic vesicles. Only triggered under intense stimulation,
not during normal stimulation. Capable of sustained release for much longer period.
• Recycling pool: (10-15%), Maintains release over time with moderate stimulation. Refilled
continuously by newly recycled vesicles.
• Readily releasable pool (RRP): Only a few vesicles (1%), already docked at active zone,
primed for release. Immediately available upon stimulation. Rapidly depleted. Responsible for
the immediate release of neurotransmitters. Fastest kinetics.
• For replenishment, you need source of new vesicles. RRP replenishes from vesicles in
recycling pool. Recycling replenished from vesicles from reserve pool.
Vesicle Recycling
• After fusion pore opens, synaptic vesicles recycle via three alternate pathways.
• Fast, Reuse (kiss-and-stay): NT uptake —> ATP —> Priming —> Ca2+ —> Pore fusion
Fastest. Vesicle never undocks from active zone, undergoes refilling with NT
• Reuse (kiss-and-run): NT uptake —> Docking —> ATP —> Priming —> Ca2+ —> Fusion pore
opening —> Ca2+ —> Endocytosis —> H+ —> Repeat
Local recycling, vesicle is endocytosed, enters recycling pool, refills, and re-docks
• Endosomal recycling: NT uptake —> Docking —> ATP —> Priming —> Ca —> Fusion pore
opening —> Ca —> Endocytosis —> H+ —> Endosome fusion —> Budding —> Repeat
Slowest. Undocked vesicle enters endosome, refills with NT, enters recycling pool.
Quantal Release
• (1936) Henry Dale, Otto Loewi prepared neuromuscular junction, and demonstrated
1. Stimulating motor nerves released acetyl choline (ACh)
2. ACh injected into arteries supplying blood to muscles caused large synchronous contraction
Bypassed motor nerve entirely, injected ACh caused twitching
3. Changes in muscle Vm by stimulation of motor nerves were mimicked by applying ACh
4. Curare antagonized these changes in membrane potential (Blocked ACh)
5. Eserine (prevents hydrolysis of ACh) potentiated these changes in membrane potential
• Showed that motor neurons released ACh in pre-synaptic terminal, and that ACh were
responsible for changes at end plates
• Neuromuscular junction: Chemical synapse at axon of motor neuron and motor end plate.
• At synaptic terminal, motor neuron releases NT (Acetylcholine (ACh)) into synaptic cleft
• Ruffled sarcolemma structure of motor end plate in order to maximize surface area available
for ACh receptors and esterases.
• Receptors are nicotinic receptors on motor end plate, binding changes ionotropic
configuration so the pore is permeable to small ions (Ca2+, Na+, K+, Cl-)
• Total ionic current is comprised mainly Na+ current because of the larger electrochemical
gradients acting on Na+ at rest. So it mainly acts as a sodium channel
• Voltage-gated sodium channels are located in the troughs of the ruffles, ACh receptor is
located on the peak of the ruffle.
• ACh receptor is anchored in place, ACh-esterase is also tethered in place nearby
• Voltage clamp recordings at the motor end plate in order to understand the cause of EPP
• Synaptic currents recorded at membrane potentials between -100 mV and +55 mV
Below 0 mV current flows into muscle. Above 0 mV, current flows out of muscle
• Final permeability is 0 mV. However, no ions have reversal potential at 0.
• Effect of ACh changes the post-synaptic membrane to change permeability of cations, results
in total current with reversal potential at 0 mV
• A. and N. Takeuchi changed the concentrations of Na, K, and Ca in the external solution
Concluded that the effect of ACh produced a general increase in cation permeability
Quantal release of neurotransmitter
• Before activating motor neuron, or applying ACh. They notice spontaneous depolarizations
which are called miniature end plate potentials (MEPPs)
MEPPs are always the same size
• In low Ca2+, unable to trigger EPP, but still MEPPs. Thus, MEPPs did not come from muscle
• MEPP produces 1 mV depolarization
• Total EPP is always a multiple of the size of the MEPP
• MEPP is the release of a single ACh vesicle which is about 10,000 molecules
• 1 vesicle = 1 quanta
• Membrane studded with population of proteins involved in docking, priming, and exocytosis
• Membrane fusion is a spontaneous process, thus fusion is not energetically favorable
• SNARE complex: SNARE proteins on vesicles (Synaptobrevin, Synaptotagmin) that interact
with SNARE proteins on membrane (Syntaxin, SNAP-25). These proteins bridge vesicle and
plasma membrane, and mediates vesicle docking/targeting.
• Synaptotagmin: Calcium sensor. Anchored to vesicle
Priming step activates Synaptotagmin to make it sensitive to calcium
Calcium binding triggers conformation in SNARE complex
• Docking and priming result from SNARE proteins (anchor vesicle, calcium sensitivity, fusion)
1. Free SNARES on vesicle and plasma membrane
2. SNARE complexes form as vesicle docks
3. Synaptotagmin binds to SNARE complex
4. Entering Ca2+ binds to synaptotagmin, curves membrane which brings membranes together.
5. Fusion of membranes leads to fusion pore, and exocytotic release of neurotransmitter
• Botulinum and tetanus toxins affect SNARE proteins involved in vesicle fusion
Multiple elements (BoTX-A, BoTX-C, BoTX-G, etc)
Each one is a different protease that targets specific cleavage site
Both of these act by blocking presynaptic membrane fusion, disrupt vesicular release
• Vesicle cycle is subject to congenital, and environmental problems. Anything interrupting
vesicle cycle, interrupts synaptic communication.
• SNARE proteins are neurotoxin targets (Botulinum and tetanus). Both of these act by
blocking presynaptic membrane fusion. Proteases that cleave presynaptic proteins.
Neurotransmitter Definitions
• Definition: Substance that is released at a synapse of by one neuron and that affects another
cell, either neuron or effector organ, in a specific manner.
• Hurley definition: First messengers released from presynaptic terminals, that must interact with
specific receptors on their target cells in order to support transmission of information.
• Information transfer occurs between either two neurons, a neuron and target effector (i.e.
skeletal muscle), or a neuron and glial cells
• Change electrical properties of target membranes by altering membrane permeability
Neurotransmitter Criteria
Neurotransmitters Categories
• Stronger signal —> Greater calcium influx —> More synaptic vesicles released
• Ca2+ release dependent on AP frequency (threshold vs supra-threshold signals)
Timeline
• NT release —> Receptor binding —> Ion channel open/close (permeability) —> Conductance
change —> Current (ion) flow —> Postsynaptic potential changes —> Postsynaptic excited/
inhibited —> Summation determines AP occurrence
• Linked to, or are ion channels. Contain two functional domains. Extracellular domain that
binds neurotransmitters. Membrane spanning domain that forms hydrophilic pore
• This pore prevents ion interaction with hydrophobic core of membrane
• Combines transmitter binding, and channel functions into single molecular entity
• Made up of multimers of individual protein subunits, each contributing to pore
• At least two state are open and closed (May be more like inactivated)
Transition between two states relies on ligand binding to receptor
• Opposite is possible —> binding of ligand causes channel to close instead of open
• Neurons and muscles both express ligand-gated channels activated by neurotransmitters
• Nicotinic-ACh receptor is a good example of ionotropic receptors
• Binding domain is combined with ion channel
• Inhibitory Postsynaptic Potential (IPSP): An inhibitory impulse, lowers the membrane potential
below rest. Hyperpolarization. Due to opening chloride channels (negative charge flows into
cell) or potassium leaving the cell.
Summation of Potentials
• Each cell receives multiple inputs, could be a combination of both EPSP and IPSP
Must be integrated by the cell to produce some response
• Two inputs in a row, without degrading, will sum together in size. This can cause an action
potential. An EPSP and IPSP in a row can cancel out.
• Temporal summation: Single EPSP (E1) input activated in close enough time that they sum
together.
• Spatial summation: Two different EPSPs (E1, E2) inputs activated in close enough time so
they sum together
• EPSP-IPSP cancellation: Two different inputs (E1, I1) activated, but one is excitatory, and
the other inhibitory.
• Large amount of integration takes place as multiple excitatory and inhibitory inputs are
activated at different times.
• Patterns of action potentials reflect patterns of input activation
• Depends on passive properties, further current spread allows great opportunity for summation
Learning Objectives (Weeks 2-4)
Excitable cells: Ability to rapidly and reversely, reverse Vm. Membrane potential is dynamic.
Non-excitable cells: Maintains Vm = Vrest at all times.
Membrane Potential (Vm): Voltage difference across membrane at any given point.
Resting Membrane Potential (Vrest): Membrane potential with no disturbances to cell.
Potential measured using glass microelectrodes inserted into the cell and extracellular fluid, and
a voltage meter recording the difference between inside and outside. Intracellular potential is
measured with respect to extracellular potential being reference point (equal to zero). The
equation used is Vm = Vi - Vo
3) Explain what is meant by the statement: “The plasma membrane is polarized.” Define
depolarization, repolarization, and hyperpolarization.
Membrane potential is typically negative with respect to outside. Therefore membrane has
asymmetry of charges across membrane. Because of this, membrane is considered polarized
Vm is maintained by Na+-K+ ATPase activity which maintains [K+]i and [Na+]i, and two organ
systems: the Kidneys and Endocrine system which maintain [K+]o and [Na+]o
Nernst equilibrium potential occurs at Vm when no net ion movement occurs because electrical
gradient balances out chemical gradient. Opposite charges attract each other on the same side,
and same charges repel each other on opposite side. No driving force.
6) Calculate the membrane potential for a neuron using the GHK equation
DFX = Vm - VX
Driving Force is the electrochemical force acting on ion if Vm =/= Veq. Causes net movement of
ions. Magnitude of DF indicates how far ion is from equilibrium. Acting on every ion. Arithmetic
sign of DF acting on ion along with valence (cation or anion) are used to predict direction of ion
flow. DF acting on different ions are not equal.
Cation (Na+, K+, Ca2+) rules: VDF > 0 then ion flow outward. VDF < 0 then ion flows inward
Anion (Cl-) rules: VDF > 0 then ion flow inward. VDF < 0 then ion flows outward.
8) Provide examples for why maintaining a negative membrane potential is important and
explain these examples
Membrane capacitance: Capacitance across membrane wall, separates charge, and determines
time constant.
Membrane resistance: Resistance across membrane wall, describes extent of current leaking
through membrane. Connected in parallel with capacitor.
Axial resistance: Resistance within axoplasm. Connected in series.
Input resistance: Reflects extent to which membrane channels are open. Decreases with larger
cell size, and greater ion channel density.
12) Explain significance of length, and time constants. What factors influence these?
Length constant (λ): Distance at which ΔVm has decayed 37% of its value compared to point of
injection. Measures how far current can flow. λ = (Rm/Ri)1/2
Time constant (τ): Time taken to reach 63% of final voltage. τm = RmCm
13) Explain the molecular and ionic events underlying each phase of the action potential
14) How was the voltage clamp method used to study how ionic currents flow during an
action potential? What evidence was presented that the early phase was carried by
sodium and the late phase by potassium?
Voltage-clamp: Studies voltage- and time-dependence of squid giant axon permeability. Analysis
demonstrated there two different, voltage-dependent permeability pathways. One, Na-selective,
which transiently activated and inactivated. Another, K-selective, which slowly activated.
Tetrodotoxin (TTX): Blocks voltage gated sodium channels. Removed transient inward current
Tetraethylammonium (TEA): Blocks voltage gated potassium channel. Removed delayed
outward current.
15) Explain the gating mechanisms for voltage gated sodium channels and potassium
channels during the action potential.
16) Describe the characteristic features of the action potential and their significance
Action potentials: Depolarize. All-or none. Frequency proportional to stimulus strength. Large
amplitude that does not decay. Duration varies with strength of stimulus. Mediated by voltage-
gated sodium and potassium channels. Reverses membrane polarity. Absolute and relative
refractory periods. Requires threshold to be reached. Caused by response to depolarizing
stimulus, and involves coordinated activity of voltage-gated ion channels.
18) Explain the difference between an absolute and relative refractory period. What is the
physiological significance of these refractory periods?
Absolute refractory period (ARP): Initiation to after peak of action potential. Another stimulus (no
matter how strong) won’t lead to second AP due to inactivated voltage-gated calcium channels.
Relative refractory period (RRP): Occurs until sodium channels recover. If strong enough stimuli
are given, cell may generate another action potential. However, stimuli must be stronger than
what was originally needed at Vrest.
Action potentials always produce the same magnitude of depolarization, thus information
encoded in another form. Stronger signals produces higher AP frequency. These action
potentials are separated by either ARP or ARP + RRP depending on signal strength.
20) Explain how action potentials are propagated down an axon and the significance of
myelination
Action potentials occur at axon hillock, and conducted down the axon. However, core (salt
solution) has poor conductance, and insulator (membrane) has low resistance. Therefore signal
can not travel as far. Myelin has high resistance which allows signal to travel farther. Axons are
typically myelinated with nodes of Ranvier in-between segments of myelin. Action potential
generates wave of depolarization down axon, followed by repolarization.
21) Describe the salient differences for electrical and chemical synapses
Electrical Synapses: Gap junctions. Links cytosol of two neighbor, Large conductance. No
synaptic delay. Reliable. Fast. Cell synchronization. Direct transfer of molecules. Capable of
modulation. Information flows in both directions. Less flexible. Less powerful.
22) Identify the chemical classes of neurotransmitters and the criteria used to establish a
particular chemical messenger as a neurotransmitter
Class II - Biogenic amines. Derived from specific amino acids. Includes catecholamines
Class III - Amino acids. Ex. Glycine, GABA (inhibitory), Glutamate (excitatory)
Neuropeptides: Transmitters made up of chains of amino acids, vary in length. Rely on protein
synthesis, they begin their life cycle in the soma, precursors must be packaged into synaptic
vesicles transported down axon to presynaptic terminals. Synthetic enzymes.
Neurotransmitters is still synthesized and packaged in presynaptic terminal.
Criteria:
4. Deactivation mechanism
23) Describe the different vesicle pools in terms of location, kinetics, size, inter-mixing
Reserve pool: (80-90%), Deposit of synaptic vesicles. Only triggered under intense stimulation,
not during normal stimulation. Capable of sustained release for much longer period.
Recycling pool: (10-15%), Maintains release over time with moderate stimulation. Refilled
continuously by newly recycled vesicles. Replenishes from reserve pool.
Readily releasable pool (RRP): (1%), Already docked at active zone, primed for release.
Immediately available upon stimulation. Rapidly depleted. Responsible for the immediate
release of neurotransmitters. Replenishes from recycling pool.
24) Explain the mechanisms of vesicle fusion. What proteins are present at the active
zone and play a role is vesicle fusion?
Excitatory Postsynaptic Potential (EPSP): An excitatory impulse, raises the membrane potential
above rest. Depolarization. Due to opening sodium channels (positive charges flow into cell)
Inhibitory Postsynaptic Potential (IPSP): An inhibitory impulse, lowers the membrane potential
below rest. Hyperpolarization. Due to opening chloride channels (negative charge flows into
cell) or potassium leaving the cell.
Temporal summation: Single EPSP (E1) inputs activated rapid enough so they sum.
Spatial summation: Two different EPSPs (E1, E2) inputs activated rapid enough so they sum.
27) Use elements of electrical circuits to model the electrical behavior of a neuron
• 3 parts - first part is short answer (one, or two sentences), 2nd is identification, 3rd is short
essay (out of five, choose two)
• Muddy points are potential essay questions
1. Describe the pathway for blood in the brain, and significance of circle of willis
2. What is the relationship between the two equations V=IR and R=1/g. Can you explain the
relationship between conductance, resistance, and a change in voltage. Explain voltage clamp
and the graph of the data presented in class.
3. Explain an action potential, and permeabilities
•
Things to do —
3. Reviewing the phases of the action potential. Describe each phase, changes of permeability,
state of gates, underlying conductance changes (timing), and how that affects shape of action
potential
3. Draw an equivalent circuit of a neuron; include passive and active elements. Be able to
explain each of these elements.