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Explore methodologies like single unit recording, extracellular recording, intracellular recording, and more in behavioral neuroscience to study brain and behavior connections.
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Methodology in Behavioral Neuroscience Jeremy L. Loebach NIH Postdoctoral Fellow Indiana University
Goals of Behavioral Neuroscience • Examine link between brain and behavior • Brain • Behavior • How?
Single Unit Recordings • Microelectrode • Glass micropipette • Conductive wire • Neutral solution • Place in or near neuron and record the electrical activity • Precise enough to isolate individual neurons
Extracellular Recording • Microelectrode • Very small tip (3-10 micron ~ 1/1,000,000 m) • Small enough to isolate a single neuron • Place near cell and record changes in electrical activity • Dendrites • EPSP • IPSP • Place near a node of Ranvier • Action potential
Receptive Fields of Simple Cells Hubel & Wiesel movie
Intracellular Recording • Microelectrode • Glass micropipette • Much smaller tip (<1 micron) • Small enough to penetrate the cell wall • Insert electrode inside of the neuron • Record changes in resting potential • Tells you about the currents entering and exiting the neuron • Change voltage of the cell and see how the cell reacts
The Voltage Clamp • Control the electrical charge of the cell from the inside • Modulate resting potential to see what effect it has on the cell • Increase • Decrease • Observe what other areas are stimulated as a result
Single Unit Recording • Pro • Can record from single neurons • Can be done in vivo or in vitro • Can systematically manipulate the conditions under which the cell will respond • Con • Invasive • Anesthesia • Difficult to do while animal is awake and behaving • Requires responses from a large number of neurons to study a system
Multi-Unit Recording • Macro electrode • Larger diameter electrode is used • Record the responses of a large number of neurons at the same time • Local field potentials • Changes in the resting potential of the neurons at the dendrites • Dipole
Multi Unit Recording • Pro • Can record many neurons at a time • Not as invasive • Can utilize awake behaving preparations • Con • Not as precise as single unit recording • Traces can include artifacts not related to the behavior
Electro Encephalography (EEG) • Recording of Local Field Potentials from the surface of the scalp • LFPs spread out and can be measured from the surface of the skin • Record electrical activity • Electrode array (1-300 channels) • Reference electrodes on neutral areas where there are no neurons • Nose • Reference for eye movements and blinks • Eye
Event Related Potential (ERP) • Recording of Local Field Potentials from the surface of the scalp • EEG • Synchronized to the onset of a stimulus • Change in electrical activity in response to an event
ERP Components • P1/N1 dipole • Initial positive peak followed by a negative peak • Sensory processing • P200 • Attention • P300 • Rarity • Oddball paradigm • N400 • Semantic processing
EEG/ERP • Pro • Noninvasive • Inexpensive • Subject does not have to be performing a task • Great timing information • Con • Spatially poor • Imprecise
Positron Emission Tomography (PET) • Measures metabolic needs of the brain • Cells require energy to function • ATP is the primary source of energy in the brain • Glucose is changed into ATP and used for energy • Follow the glucose to see which areas are active during a task
PET • How? • Attach a radioactive isotope to the glucose • Cyclotron generates the isotope • Short half life (1 hour or less) • When decays it emits a positron • Positron collides with an electron and explodes, releasing photons at collision site • Photosensitive ring detects the photons • Triangulate the source • Register to the brain
PET • Are not limited to glucose alone • Can attach tracer to any molecule • Neurotransmitter • See where a neurotransmitter is being used • Parkinson’s • Dopamine • DOPA is a precursor for Dopamine • Converted to Dopamine in substantia nigra • Substantia nigra is destroyed, Dopamine production decreases • Trace DOPA
PET • Pro • Any molecule can be tagged • Con • Expensive • Injecting the body with radioactive substance • Poor resolution
Functional Magnetic Resonance Imaging • Blood Oxygen Level Dependent Signal (BOLD) • Track oxygen in the blood using magnets • Oxygenated blood is high in iron • Iron is magnetic • Oxygen is also used by cells • When Oxygen is stripped from the blood by the cells, the blood becomes less magnetic • Flows back to the heart and lungs for re-oxygenation
fMRI • Pro • Great spatial resolution • Relatively inexpensive • Con • Poor temporal resolution • 1-5 seconds • Indirect measure of neural activity • Not sure what governs the hemodynamic response in the first place
Event Related Optical Signal (EROS) • Uses light to assess function • Skull is transparent to light • Shine light through skull at an angle, it will arc and exit elsewhere • Depending on what gets in the way, the light will be differentially deflected • Measure the amount of light that returns to assess neural function • Swelling of neuron due to ions entering and leaving causes light to be deflected
EROS • Pro • Great temporal resolution • Milliseconds • Fast • Inexpensive • Con • Requires registration to MRI • Surface --- can only reach a few centimeters below cortical surface
Summary • Depending on what you are looking for, some methods may be more appropriate than others • Know the strengths of a technique • Know the limitations of a technique • Modern neuroimaging allows extension of techniques used in animals to humans • Ask the right questions • Use the right methodology
How do we observe this experimentally? • Record the activity of individual neurons during a task to see what they do • Extracellular • Intracellular • Sample the extracellular fluid from the synapse • What neurotransmitters are being used • Block the receptor and see how behavior is affected • Record from multiple neurons to see what a given brain region is responsible for
Review: Anatomy of the Neuron • Dendrites • Soma • Axon • Nodes of Ranvier • Terminal Boutons
Review: Neurotransmitters • Neurotransmitter • Chemical substances used in neural communication • Made in soma • Packaged in vesicles • Transported down the axon to the terminal boutons
Review: Neurotransmission • Arrival of AP stimulates release • Crosses synaptic cleft to stimulate post-synaptic neuron