02 May 2009

Functional Neuroanatomy



Functional Neuroanatomy

Histology
Neurons and Glial Cells
Brain Cell Structure and Categorization
Synapse Function
Axonal Transport
Information Flow
Glial Types and Functions

Histological Techniques
* Golgi Stain
* Nissl Stain
* Immunocytochemistry
* Track-tracing (horseradish peroxidase)

Neurons and Glials

* Brain Cells – 2 types
* Characterization
o Shape
o Size
o Function
* Organization

Neuron Components

* Dendrites
* Soma
* Axon Hillock
* Axon
* Terminal Button(s) (Bouton)

Neuron Characterization

* Shape
* Size
* Function

* Monopolar
* Bipolar
* Mutipolar

Neuron Size
Neuron Function
* Sensory
* Motor
* Interneuron
Basis of the Synapse

* Pre-synaptic Zone
* Post-synaptic zone
* Synaptic Cleft
* Synaptic Vesicles
* Dendritic Spines (size & number can be altered)
* Receptors
Axonal Transport
* Afferents (toward area) / efferents (away from area)
* Saltatory Transport (along microtubules, neurofilaments & microfilaments)
* Anterograde
* Retrograde

Glial Function
Glial Types
myelin-producing

Functional Neuroanatomy.ppt

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Research Methods in Biopsychology



RESEARCH METHODS IN BIOPSYCHOLOGY

LECTURE NOTES
METHODS TO STUDY THE NERVOUS SYSTEM

* Brain Imaging & Brain Stimulation Techniques in Living Humans
* Psychophysiological Techniques
* Invasive Physiological Methods
* Neuropharmacological Methods
* Genetic Engineering

BEHAVIORAL METHODS IN BIOPSYCHOLOGY
* Neuropsychological Testing
* Behavioral Methods in Cognitive Neuroscience
* Animal Behavior Paradigms

IMAGING & STIMULATING THE LIVING BRAIN
* Contrast X-rays
o cerebral angiography
* Computed Tomography (CT)
* Magnetic Resonance Imaging (MRI)
* Positron Emission Tomography (PET)
* Functional MRI (fMRI)
* Magnetoencephalography (MEG)
* Transcranial Magnetic Stimulation (TMS)

CT SCANS
* A computer assisted X-ray procedure
* An X-ray scanner is rotated 1o at a time over 180 o
* Computer reconstruction
* Horizontal sections
* Reveal structural abnormalities, such as cortical atrophy or lesions caused by a stroke or trauma.

MRI SCANS
* A strong magnetic field causes hydrogen atoms to align in the same orientation.
* When a radio frequency wave is passed through the head, atomic nuclei emit electromagnetic energy.
* The MRI scanner is tuned to detect radiation emitted from the hydrogen molecules.
* Computer reconstructs image.

MRI VS. CT SCANS
* Advantages of MRI
o No ionizing radiation exposure
o Better spatial resolution
o Horizontal, Frontal or Sagittal planes
* Disadvantages
o Cost
o No ferrous metal!

PET SCANS
* A positron emitting radionuclide is injected (e.g., 2-deoxyglucose).
* Positrons interact with electrons which produce photons (gamma rays) traveling in opposite directions.
* PET scanner detects the photons.
* Computer determines how many gamma rays from a particular region and a map is made showing areas of high to low activity.

PET Versus CAT

* CAT scans show brain structures.
* PET scans reveal brain activity.
* CAT involves absorption of X-rays.
* PET involves emission of radiation by an injected or inhaled isotope.

FUNCTIONAL MRI
* Images brain hemodynamics.
* Advantages over PET:
o No injections need to be given
o Structure and Function
o Shorter imaging time
o Better spatial resolution
o 3-D images

Magnetoencephalography (MEG)

* MEG measures changes in magnetic fields on the scalp surface that are produced by changes in patterns of neural activity.
* Advantage over fMRI
o faster temporal resolution
* Advantage over EEG
o greater accuracy and more reliable localization due to minimal distortion of the signal
* Clinical Uses
o Evaluation of epilepsy: to localize the source of epileptiform brain activity, usually performed with simultaneous EEG

Transcranial Magnetic Stimulation

* TMS disrupts neural activity by creating a magnetic field under a coil positioned near the skull.
o Disruption of specific cortical locations are produced while participants engage in cognitive and/or behavioral tasks.
o This allows researchers to assess functions of specific cortical areas.

PSYCHOPHYSIOLOGY
* Electroencephalography (EEG)
* Electromyography (EMG)
* Electrooculography (EOG)
* Electrodermal activity (Skin Conductance)
* Cardiovascular activity
o Heart rate (EKG)
o Blood Pressure
o Plethysmography

INVASIVE PHYSIOLOGICAL METHODS IN NONHUMANS
* Stereotaxic Surgery
* Lesion Methods
* Electrical Stimulation
* Electrophysiological Recording

LESIONING TECHNIQUES
* Aspiration lesions
* Radio-frequency lesions
* Knife cuts
* Cryogenic blockade
* Chemical Lesions

NEUROHISTOLOGY TECHNIQUES

* Fixation, preservation of tissue, sectioning and staining of tissue
* Uses of histological techniques
o Confirming lesion sites or electrode locations
o In combination with neural tracing techniques (anterograde, retrograde labeling)
o In combination with autoradiography or immunohistochemistry techniques

NEUROHISTOLOGICAL STAINING TECHNIQUES
* Nissl Stains
o e.g., cresyl violet
o stains mainly cell bodies
* Golgi Silver Stain
o stains whole neurons
* Myelin Stains (Fiber stains)
o e.g., Weigert stain
o stains mainly myelin

ELECTROPHYSIOLOGY TECHNIQUES

* Intracellular unit recording
* Extracellular unit recording
* Multiple-unit recording
o See page 114 in Pinel

NEUROPHARMACOLOGICAL METHODS Measuring Chemical Activity in the Brain

* 2-DG Autoradiography
o Radioactive 2-deoxyglucose is injected
o Animal engages in behavior of interest
o Animal is euthanized, brain tissue is removed and sliced
o Tissue slices are coated with photographic emulsion and stored in the dark (much like film processing)
o Areas that absorbed high levels of radioactive substance will appear darker
o Using computer imaging, differences in density can be color coded.
+ e.g., see page 115 in Pinel
* Cerebral Dialysis (in vivo microdialysis)
o Under anesthesia and stereotaxic guidance, a cannula is inserted into a specific brain site.
o Following recovery, a small probe with a semipermeable membrane is inserted into the cannula.
o Fluid is perfused through the probe and chemicals in the extracellular fluid diffuse across the membrane and are collected into a sample vial.
o The samples are then analyzed using a chromatography methods. (e.g. HPLC)

NEUROPHARMACOLOGICAL METHODS
Localizing Neurotransmitters and Receptors

* Immunocytochemistry
o Makes use of antibodies for specific proteins, such as receptors or enzymes.
o The antibody is labeled with a fluorescent die or a radioactive element (commercially available).
o Brain tissue is sliced and exposed to a solution containing the labeled antibody.
o Brain slices are viewed under microscope to identify the regions where protein of interest is distributed.
* In situ hybridization
o Also used to locate peptides or proteins in tissue
o Hybrid strands of mRNA are artificially created and labeled with a dye or radioactive element
o Brain tissue slices are exposed to solution containing the labeled mRNA
o Brain slices are viewed under microscope to identify regions where the mRNA expression is highest
+ e.g., see page 117 in Pinel

GENETIC ENGINEERING
ANIMAL BEHAVIOR PARADIGMS

RESEARCH METHODS IN BIOPSYCHOLOGY.ppt

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Methods and Strategies of Research



Methods and Strategies of Research
Copyright 2001 by Allyn & Bacon

Experimental Ablation

* Ablation involves the destruction of brain tissue followed by an assessment of subsequent changes in behavior
* Ablation techniques include
o Electrolytic lesions/Radio Frequency lesions
o Excitotoxic lesions (kainic acid)
o Neurochemical lesions (6-OHDA)
o Aspiration
o Knife cuts
* Distinction between functions and behaviors
* Brain lesion studies are complicated by the fact that all regions of the brain are interconnected



Stereotaxic Surgery

* A stereotaxic instrument holds the head in a fixed position
o The instrument has an arm that can move in 3 dimensions
o The surgeon can thus position an electrode or other device within a particular sub-cortical structure
* A stereotaxic atlas provides a series of drawings of brain structures
o Each page is a section of brain relative to a landmark on the skull (such as bregma)

Using a Stereotaxic Atlas to Target a Brain Lesion

Histological Techniques

* Histological techniques are used to verify the placement of a lesion within brain
o Perfuse (to remove blood from brain)
+ Remove brain
o Fix brain in formalin to solidify tissue and to prevent autolysis
+ Slice brain into thin sections (10-80 microns thick)
o Use stains to highlight selective neural elements
+ Myelin (Weil stain)
+ Cell body (cresyl violet: Nissl substance in cytoplasm)
+ Membrane (Golgi stain)

Defining Neural Connections

* Neurons in a given region send axonal outputs (efferents) to other brain regions and receive axonal inputs (afferents)
o Tracing efferent connections is done using anterograde labels that are taken up by the cell bodies and transported to axons
+ “Forward: toward axons from cell bodies”
+ Inject the lectin PHA-L into a nucleus, wait several days, process brain tissue.
+ Immunocytochemistry uses a radioactive antibody to PHA-L in order to identify cells containing PHA-L
o Tracing afferent connections is done using retrograde labeling
+ “Backwards: from axons to cell bodies”
+ e.g. fluorogold is a retrograde tracer

Anterograde Tracing
Visualizing a Living Human Brain
Human MRI (Normal)
Images courtesy of Dr. Nancy Andreason
Recording Neural Activity

* Axons conduct action potentials and neurotransmitters elicit postsynaptic potentials
* The electrical events of a discrete region can be recorded using glass microelectrodes (acute recording) or tungsten wire (chronic recording)
* Macroelectrodes record the summated electrical activity of large regions of brain
o Surface electrodes placed on human scalp are used to record brain activity (electroencephalogram: EEG)

Recording Synaptic Activity
Human Brain Imaging
Microdialysis
Microdialysis Probe Details
Artificial Stimulation of Brain
Localization of Neurotransmitters
Receptor Localization Techniques

Genetic Methods
* Genetic research methods seek to demonstrate the linkage between genes and behavior
* Twin studies examine the impact of varying degrees of genetic similarity on behavioral similarity
o Identical twins (MZ) share 100% of their genes while fraternal twins (DZ) share about 50% of their genes
o Concordance rate examines the likelihood of whether a twin shares a behavioral trait with the other twin
o A higher concordance rate for MZ twins relative to DZ twins suggests a genetic influence for that characteristic
* Adoption studies examine the similarity with regard to a trait for an adopted person compared to their adopted parents and their biological parents
* Targeted mutations involve the insertion of defective (knockout) genes into the chromosomes of mice

Methods and Strategies of Research.ppt

Original material copyright ©1997-2005 Addiction Research Unit. Original material presented here may be reprinted or used in any non-commercial form if proper credit is given to source and if original intent remains intact. All other material remains property of its copyright holder and is protected by international copyright laws.

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