Name: investigating the neural mechanisms of aware and unaware fear memory with fmri
Uploaded: 2011-10-06T12:00:00
Description: Watch this Scientific Journal Video about Investigating the Neural Mechanisms of Aware and Unaware Fear Memory with fMRI at JoVE.com

Support provided by the University of Alabama at Birmingham Faculty Development Grant Program.

1. Psychophysiology
The Biopac Systems, Inc. physiological monitoring system (see Table of specific equipment) is non-standard equipment in most imaging facilities. Schedule 15-30 minutes prior to participant arrival to set up physiological monitoring and other equipment described in this protocol (Figure 1).
<ol>
<li>Connect a control room computer operating AcqKnowledge (Biopac Systems, Inc.) physiological monitoring software to the Biopac MP150 (MP150WSW) using a standard Ethernet crossov...

<ol>
	<li>LaBar, K. S., Gatenby, J. C., Gore, J. C., LeDoux, J. E., Phelps, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+amygdala+activation+during+conditioned+fear+acquisition+and+extinction:+a+mixed-trial+fMRI+study.">Human amygdala activation during conditioned fear acquisition and extinction: a mixed-trial fMRI study.</a> Neuron. 20, 937-945 (1998).</li><li>Buchel, C., Morris, J., Dolan, R. J., Friston, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+systems+mediating+aversive+conditioning:+an+event-related+fMRI+study.">Brain systems mediating aversive conditioning: an event-related fMRI study.</a> Neuron. 20, 947-957 (1998).</li><li>Cheng, D. T., Knight, D. C., Smith, C. N., Stein, E. A., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+MRI+of+human+amygdala+activity+during+Pavlovian+fear+conditioning:+stimulus+processing+versus+response+expression.">Functional MRI of human amygdala activity during Pavlovian fear conditioning: stimulus processing versus response expression.</a> Behav. Neurosci. 117, 3-10 (2003).</li><li>Knight, D. C., Smith, C. N., Stein, E. A., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+MRI+of+human+Pavlovian+fear+conditioning:+patterns+of+activation+as+a+function+of+learning.">Functional MRI of human Pavlovian fear conditioning: patterns of activation as a function of learning.</a> Neuroreport. 10, 3665-3670 (1999).</li><li>Cheng, D. T., Knight, D. C., Smith, C. N., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+amygdala+activity+during+the+expression+of+fear+responses.">Human amygdala activity during the expression of fear responses.</a> Behav. Neurosci. 120, 1187-1195 (2006).</li><li>Balderston, N. L., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conditioning+with+masked+stimuli+affects+the+timecourse+of+skin+conductance+responses.">Conditioning with masked stimuli affects the timecourse of skin conductance responses.</a> Behav. Neurosci. 124, 478-489 (2010).</li><li>Esteves, F., Parra, C., Dimberg, U., Ohman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonconscious+associative+learning:+Pavlovian+conditioning+of+skin+conductance+responses+to+masked+fear-relevant+facial+stimuli.">Nonconscious associative learning: Pavlovian conditioning of skin conductance responses to masked fear-relevant facial stimuli.</a> Psychophysiology. 31, 375-385 (1994).</li><li>Cheng, D. T., Richards, J., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity+in+the+human+amygdala+corresponds+to+early,+rather+than+late+period+autonomic+responses+to+a+signal+for+shock.">Activity in the human amygdala corresponds to early, rather than late period autonomic responses to a signal for shock.</a> Learn. Mem. 14, 485-490 (2007).</li><li>Knight, D. C., Nguyen, H. T., Bandettini, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+human+amygdala+in+the+production+of+conditioned+fear+responses.">The role of the human amygdala in the production of conditioned fear responses.</a> Neuroimage. 26, 1193-1200 (2005).</li><li>Knight, D. C., Nguyen, H. T., Bandettini, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+awareness+in+delay+and+trace+fear+conditioning+in+humans.">The role of awareness in delay and trace fear conditioning in humans.</a> Cogn. Affect. Behav. Neurosci. 6, 157-162 (2006).</li><li>Schultz, D. H., Helmstetter, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classical+conditioning+of+autonomic+fear+responses+is+independent+of+contingency+awareness.">Classical conditioning of autonomic fear responses is independent of contingency awareness.</a> J. Exp. Psychol. Anim. Behav. Process. 36, 495-500 (2010).</li><li>Dunsmoor, J. E., Bandettini, P. A., Knight, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+correlates+of+unconditioned+response+diminution+during+Pavlovian+conditioning.">Neural correlates of unconditioned response diminution during Pavlovian conditioning.</a> Neuroimage. 40, 811-817 (2008).</li><li>Katkin, E. S., Wiens, S., Ohman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonconscious+fear+conditioning,+visceral+perception,+and+the+development+of+gut+feelings.">Nonconscious fear conditioning, visceral perception, and the development of gut feelings.</a> Psychol. Sci. 12, 366-370 (2001).</li><li>Knight, D. C., Waters, N. S., King, M. K., Bandettini, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Learning-related+diminution+of+unconditioned+SCR+and+fMRI+signal+responses.">Learning-related diminution of unconditioned SCR and fMRI signal responses.</a> Neuroimage. 49, 843-848 (2010).</li><li>Knight, D. C., Waters, N. S., Bandettini, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+substrates+of+explicit+and+implicit+fear+memory.">Neural substrates of explicit and implicit fear memory.</a> Neuroimage. 45, 208-214 (2009).</li><li>Lovibond, P. F., Shanks, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+awareness+in+Pavlovian+conditioning:+empirical+evidence+and+theoretical+implications.">The role of awareness in Pavlovian conditioning: empirical evidence and theoretical implications.</a> J. Exp. Psychol. Anim. Behav. Process. 28, 3-26 (2002).</li><li>Hippocampus, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> 8, 620-626 (1998).</li><li>Cox, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AFNI:+software+for+analysis+and+visualization+of+functional+magnetic+resonance+neuroimages.">AFNI: software for analysis and visualization of functional magnetic resonance neuroimages.</a> Comput. Biomed. Res. 29, 162-173 (1996).</li><li>Knight, D. C., Nguyen, H. T., Bandettini, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+conditional+fear+with+and+without+awareness.">Expression of conditional fear with and without awareness.</a> Proc. Natl. Acad. Sci. U. S. A. 100, 15280-15283 (2003).</li><li>Bunce, S. C., Bernat, E., Wong, P. S., Shevrin, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Further+evidence+for+unconscious+learning:+preliminary+support+for+the+conditioning+of+facial+EMG+to+subliminal+stimuli.">Further evidence for unconscious learning: preliminary support for the conditioning of facial EMG to subliminal stimuli.</a> J. Psychiatr. Res. 33, 341-347 (1999).</li><li>Kotze, H. F., Moller, A. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+auditory+subliminal+stimulation+on+GSR.">Effect of auditory subliminal stimulation on GSR.</a> Psychol. Rep. 67, 931-934 (1990).</li><li>Miller, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Threshold+variability+in+subliminal+perception+experiments:+fixed+threshold+estimates+reduce+power+to+detect+subliminal+effects.">Threshold variability in subliminal perception experiments: fixed threshold estimates reduce power to detect subliminal effects.</a> J. Exp. Psychol. Hum. Percept. Perform. 17, 841-851 (1991).</li><li>Tabbert, K., Stark, R., Kirsch, P., Vaitl, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissociation+of+neural+responses+and+skin+conductance+reactions+during+fear+conditioning+with+and+without+awareness+of+stimulus+contingencies.">Dissociation of neural responses and skin conductance reactions during fear conditioning with and without awareness of stimulus contingencies.</a> Neuroimage. 32, 761-770 (2006).</li></ol>

The fear conditioning methodology described here provides a means to investigate the neural mechanisms of aware and unaware fear memory processes. This method takes advantage of the simultaneous monitoring of behavioral, autonomic, and fMRI data. Monitoring behavioral (i.e. UCS expectancy) and autonomic responses (i.e. SCR) is a critical component of this method. UCS expectancy provides a means to assess contingency awareness, while SCR provides an index of CR expression. Together, these behavioral and autonomic response...

<table border="1">
<tbody>
 <tr>
 <td>Equipment</td>
 <td>Company</td>
 <td>Item number</td>
 </tr>
 <tr>
 <td>Integrated Functional Imaging System (IFIS-SA)</td>
 <td>Invivo Corp., Orlando, FL</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Master Control Unit (located in the control room) 
 Peripheral Interface Unit (located in the MRI chamber) 
 Audio/Visual Display Unit (located in the MRI chamber), includes:
 <ul>
 <li>6.4" (diagonal) LCD video screen <ul>
 <li>640 x 480 resolution and 15&deg; field of view</li></ul></li>
 <li>acoustic interface box<ul>
 <li>delivers pneumatic sound in stereo</li></ul></li>
 <li>MR-compatible stereo headphones </li>
 </ul></td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>PHYSIOLOGICAL MONITORING SYSTEM</td>
 <td>Biopac Systems, Inc., Goleta, CA</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Data Acquisition and Analysis System for Windows (MP150) 
 Isolated Digital Interface (Digital Interface) 
 Galvanic Skin Response (GSR) Amplifier 
 MRI Cable/Filter System to Transducer Amplifier set, includes: 
 <ul>
 <li>MRI extension cable (Chamber to filter)</li>
 <li>RF interference filter </li>
 <li>MRI extension Cable (GSR amplifier to filter)</li>
 </ul>
 Additional components: 
 DB25 M/F ribbon cable 
 Disposable radiotranslucent electrodes 
 Carbon fiber leads</td>
 <td>&nbsp;</td>
 <td valign="top">MP150WSW 
 STP100C 
 EDA100C-MRI 
 MECMRI-TRANS 
 - MECMRI-1 
 - MRIRFIF 
 - MECMRI-3 
 CBL110C 
 EL508 
 LEAD108
 </td>
 </tr>
 <tr>
 <td>JOYSTICK</td>
 <td>Current Designs, Inc., Philadelphia, PA</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Legacy Joystick</td>
 <td>&nbsp;</td>
 <td>HH-JOY-4</td>
 </tr>
 <tr>
 <td>Legacy fORP Interface</td>
 <td>&nbsp;</td>
 <td>FIU-005</td>
 </tr>
 </tbody>
</table>

investigating the neural mechanisms of aware and unaware fear memory with fmri

Pavlovian fear conditioning is often used in combination with functional magnetic resonance imaging (fMRI) in humans to investigate the neural substrates of associative learning 1-5. In these studies, it is important to provide behavioral evidence of conditioning to verify that differences in brain activity are learning-related and correlated with human behavior. 
Fear conditioning studies often monitor autonomic responses (e.g. skin conductance response; SCR) as an index of learning and memory 6-8. In addition, other behavioral measures can provide valuable information about the learning process and/or other cognitive functions that influence conditioning. For example, the impact unconditioned stimulus (UCS) expectancies have on the expression of the conditioned response (CR) and unconditioned response (UCR) has been a topic of interest in several recent studies 9-14. SCR and UCS expectancy measures have recently been used in conjunction with fMRI to investigate the neural substrates of aware and unaware fear learning and memory processes 15. Although these cognitive processes can be evaluated to some degree following the conditioning session, post-conditioning assessments cannot measure expectations on a trial-to-trial basis and are susceptible to interference and forgetting, as well as other factors that may distort results 16,17 .
Monitoring autonomic and behavioral responses simultaneously with fMRI provides a mechanism by which the neural substrates that mediate complex relationships between cognitive processes and behavioral/autonomic responses can be assessed. However, monitoring autonomic and behavioral responses in the MRI environment poses a number of practical problems. Specifically, 1) standard behavioral and physiological monitoring equipment is constructed of ferrous material that cannot be safely used near the MRI scanner, 2) when this equipment is placed outside of the MRI scanning chamber, the cables projecting to the subject can carry RF noise that produces artifacts in brain images, 3) artifacts can be produced within the skin conductance signal by switching gradients during scanning, 4) the fMRI signal produced by the motor demands of behavioral responses may need to be distinguished from activity related to the cognitive processes of interest. Each of these issues can be resolved with modifications to the setup of physiological monitoring equipment and additional data analysis procedures. Here we present a methodology to simultaneously monitor autonomic and behavioral responses during fMRI, and demonstrate the use of these methods to investigate aware and unaware memory processes during fear conditioning.

Watch this Scientific Journal Video about Investigating the Neural Mechanisms of Aware and Unaware Fear Memory with fMRI at JoVE.com

Investigating the Neural Mechanisms of Aware and Unaware Fear Memory with fMRI

A methodology to investigate the neural mechanisms that support aware and unaware memory processes during fear conditioning is described. This method monitors blood oxygen level dependent (BOLD) functional magnetic resonance imaging, skin conductance response, and unconditioned stimulus expectancy during Pavlovian fear conditioning to assess the neural correlates of distinct memory processes.

Pavlovian fear conditioning is often used in combination with functional magnetic resonance imaging (fMRI) in humans to investigate the neural substrates ...

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