We are grateful to Canine Performance Sciences and Auburn University Departments of Psychology and Electrical &#38; Computer Engineering. This work was supported by the Association of Professional Dog Trainers.

<ol>
	<li>T&#243;th, L., G&#225;csi, M., Mikl&#243;si, &. #. 1. 9. 3. ;., Bogner, P., Repa, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Awake+dog+brain+magnetic+resonance+imaging.">Awake dog brain magnetic resonance imaging.</a> Journal of Veterinary Behavior. 4 (2), (2009).</li><li>Berns, G. S., Brooks, A. M., Spivak, M. <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+in+awake+unrestrained+dogs.">Functional MRI in awake unrestrained dogs.</a> PLoS One. 7 (5), e38027 (2012).</li><li>Bunford, N., Andics, A., Kis, A., Miklosi, A., Gacsi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Canis+familiaris+As+a+Model+for+Non-Invasive+Comparative+Neuroscience.">Canis familiaris As a Model for Non-Invasive Comparative Neuroscience.</a> Trends in Neurosciences. 40 (7), 438-452 (2017).</li><li>Jia, H., et al. <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+the+Olfactory+System+in+Conscious+Dogs.">Functional MRI of the Olfactory System in Conscious Dogs.</a> Plos One. 9 (1), e86362 (2014).</li><li>Thompkins, A. M., Deshpande, G., Waggoner, P., Katz, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+Magnetic+Resonance+Imaging+of+the+Domestic+Dog:+Research,+Methodology,+and+Conceptual+Issues.">Functional Magnetic Resonance Imaging of the Domestic Dog: Research, Methodology, and Conceptual Issues.</a> Comparative Cognition &amp; Behavior Reviews. 11, 63-82 (2016).</li><li>Berns, G. S., Cook, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Why+Did+the+Dog+Walk+Into+the+MRI?.">Why Did the Dog Walk Into the MRI?.</a> Current Directions in Psychological Science. 25 (5), 363-369 (2016).</li><li>Huber, L., Lamm, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+dog+cognition+by+functional+magnetic+resonance+imaging.">Understanding dog cognition by functional magnetic resonance imaging.</a> Learning &amp; Behavior. 45 (2), 101-102 (2017).</li><li>Ramirez, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Animal training: successful animal management through positive reinforcement. , (1999).</li><li>Gerencser, L., Bunford, N., Moesta, A., Miklosi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+validation+of+the+Canine+Reward+Responsiveness+Scale+-Examining+individual+differences+in+reward+responsiveness+of+the+domestic+dog.">Development and validation of the Canine Reward Responsiveness Scale -Examining individual differences in reward responsiveness of the domestic dog.</a> Scientific Reports. 8 (1), 4421 (2018).</li><li>Thompkins, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Separate+brain+areas+for+processing+human+and+dog+faces+as+revealed+by+awake+fMRI+in+dogs+(Canis+familiaris).">Separate brain areas for processing human and dog faces as revealed by awake fMRI in dogs (Canis familiaris).</a> Learning &amp; Behavior. 46 (4), 561-573 (2018).</li><li>Lazarowski, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+of+the+Behavioral+Characteristics+of+Dogs+Purpose-Bred+and+Prepared+to+Perform+Vapor+Wake&#174;+Detection+of+Person-Borne+Explosives.">Investigation of the Behavioral Characteristics of Dogs Purpose-Bred and Prepared to Perform Vapor Wake&#174; Detection of Person-Borne Explosives.</a> Frontiers in Veterinary Science. 5 (50), (2018).</li><li>Lazarowski, L., Waggoner, P., Katz, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+future+of+detector+dog+research.">The future of detector dog research.</a> Comparative Cognition &amp; Behavior Reviews. 14, 77-80 (2019).</li><li>Leidinger, C., Herrmann, F., Thone-Reineke, C., Baumgart, N., Baumgart, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Introducing+Clicker+Training+as+a+Cognitive+Enrichment+for+Laboratory+Mice.">Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice.</a> Journal of Visualized Experiments. (121), e55415 (2017).</li><li>Council, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Guide for the Care and Use of Laboratory Animals. , (2011).</li><li>Andics, A., G&#225;bor, A., Farag&#243;, T., Szab&#243;, D., Mikl&#243;si, &. #. 1. 9. 3. ;. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+mechanisms+for+lexical+processing+in+dogs.">Neural mechanisms for lexical processing in dogs.</a> Science. 353 (6303), 1030-1032 (2016).</li><li>Berns, G. S., Brooks, A. M., Spivak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Scent+of+the+familiar:+An+fMRI+study+of+canine+brain+responses+to+familiar+and+unfamiliar+human+and+dog+odors.">Scent of the familiar: An fMRI study of canine brain responses to familiar and unfamiliar human and dog odors.</a> Behavioural Processes. 110, 37-46 (2015).</li><li>Berns, G. S., Brooks, A., Spivak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Replicability+and+Heterogeneity+of+Awake+Unrestrained+Canine+fMRI+Responses.">Replicability and Heterogeneity of Awake Unrestrained Canine fMRI Responses.</a> PLoS One. 8 (12), e81698 (2013).</li><li>Cook, P., Prichard, A., Spivak, M., Berns, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Jealousy+in+dogs?+Evidence+from+brain+imaging.">Jealousy in dogs? Evidence from brain imaging.</a> Animal Sentience. 117, 1-15 (2018).</li><li>Cook, P. F., Brooks, A., Spivak, M., Berns, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regional+brain+activations+in+awake+unrestrained+dogs.">Regional brain activations in awake unrestrained dogs.</a> Journal of Veterinary Behavior-Clinical Applications and Research. 16, 104-112 (2016).</li><li>Cook, P. F., Prichard, A., Spivak, M., Berns, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Awake+canine+fMRI+predicts+dogs'+preference+for+praise+vs+food.">Awake canine fMRI predicts dogs' preference for praise vs food.</a> Social Cognitive and Affective Neuroscience. 11 (12), 1853-1862 (2016).</li><li>Cook, P. F., Spivak, M., Berns, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurobehavioral+evidence+for+individual+differences+in+canine+cognitive+control:+an+awake+fMRI+study.">Neurobehavioral evidence for individual differences in canine cognitive control: an awake fMRI study.</a> Animal Cognition. 19 (5), 867-878 (2016).</li><li>Cook, P. F., Spivak, M., Berns, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=One+pair+of+hands+is+not+like+another:+caudate+BOLD+response+in+dogs+depends+on+signal+source+and+canine+temperament.">One pair of hands is not like another: caudate BOLD response in dogs depends on signal source and canine temperament.</a> PeerJ. 2, e596 (2014).</li><li>Dilks, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Awake+fMRI+reveals+a+specialized+region+in+dog+temporal+cortex+for+face+processing.">Awake fMRI reveals a specialized region in dog temporal cortex for face processing.</a> PeerJ. 3, e1115 (2015).</li><li>Prichard, A., Chhibber, R., Athanassiades, K., Spivak, M., Berns, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+neural+learning+in+dogs:+A+multimodal+sensory+fMRI+study.">Fast neural learning in dogs: A multimodal sensory fMRI study.</a> Scientific Reports. 8 (1), 14614 (2018).</li><li>Prichard, A., Cook, P. F., Spivak, M., Chhibber, R., Berns, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Awake+fMRI+Reveals+Brain+Regions+for+Novel+Word+Detection+in+Dogs.">Awake fMRI Reveals Brain Regions for Novel Word Detection in Dogs.</a> Frontiers in Neuroscience. 12, 737 (2018).</li><li>Ramaihgari, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zinc+Nanoparticles+Enhance+Brain+Connectivity+in+the+Canine+Olfactory+Network:+Evidence+From+an+fMRI+Study+in+Unrestrained+Awake+Dogs.">Zinc Nanoparticles Enhance Brain Connectivity in the Canine Olfactory Network: Evidence From an fMRI Study in Unrestrained Awake Dogs.</a> Frontiers in Veterinary Science. 5, 127 (2018).</li><li>Robinson, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+Structural+Connectivity+of+the+Default+Mode+Network+in+Dogs+using+Diffusion+Tensor+Imaging.">Characterization of Structural Connectivity of the Default Mode Network in Dogs using Diffusion Tensor Imaging.</a> Scientific Reports. 6, 36851 (2016).</li><li>Berns, G. S., Brooks, A. M., Spivak, M., Levy, K. <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+in+Awake+Dogs+Predicts+Suitability+for+Assistance+Work.">Functional MRI in Awake Dogs Predicts Suitability for Assistance Work.</a> Scientific Reports. 7, 43704 (2017).</li><li>Berns, G. S., Spivak, M., Nemanic, S., Northrup, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+Findings+in+Dogs+Trained+for+Awake-MRI.">Clinical Findings in Dogs Trained for Awake-MRI.</a> Frontiers in Veterinary Science. 5, 209 (2018).</li><li>Jia, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+odor-induced+activity+in+the+canine+brain+using+zinc+nanoparticles:+A+functional+MRI+study+in+fully+unrestrained+conscious+dogs.">Enhancement of odor-induced activity in the canine brain using zinc nanoparticles: A functional MRI study in fully unrestrained conscious dogs.</a> Chemical Senses. 41 (1), 53-67 (2016).</li><li>Kyathanahally, S. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anterior-posterior+dissociation+of+the+default+mode+network+in+dogs.">Anterior-posterior dissociation of the default mode network in dogs.</a> Brain Structure and Function. 220 (2), 1063-1076 (2015).</li></ol>

The authors have nothing to disclose.

The protocol described above separates the training of the stationing (chin rest) behavior from desensitization to the MRI environment. Further, it utilizes a generalization procedure of stationing in several dissimilar locations, to assist in the transfer of the stationing behavior to the real MRI scan environment; it does so without the need for extensive training time in the MRI scan environment, which can be expensive. Overall, the training and testing was completed in 14 hours and resulted in immediate transfer to n...

Magnetic resonance imaging (MRI) conducted on unrestrained awake dogs is a new method, creating a fresh way to examine function and structure in the dog brain. The first published accounts of MR image acquisition from unrestrained awake dogs were published in 2009 (structural) and 2012 (functional)1,2. There are several advantages of functional magnetic resonance imaging (fMRI) for studying brain function in unrestrained awake dogs. First, the data collection is similar to that of humans, and therefore more readily generalizable across species3. Second, there is no need for anesthesia, eliminating any undesirable aftereffects. Third, brain activity is altered by anesthesia and hence cognitive function can be better assessed without anesthesia4. Fourth, while fluid/food deprivation and physical restraint allow researchers to probe nonsedated animals (e.g., rodent, avian, and primate models), those animals can be in very different cognitive states from their non-deprived and unrestrained counterparts3.
At the moment, there are five laboratories around the world that are scanning awake dogs (Atlanta, USA; Auburn, USA; Budapest, Hungary; Quer&#233;taro, Mexico; Vienna, Austria), and there is no standardized method for training dogs to willfully undergo an MRI scan5,6,7. All the training methods share the common goal of training dogs to remain still for extended periods of time, which is necessary for quality brain scans. While all methods work via the principles of reinforcement learning, how exactly it is implemented varies, and we do not yet know the impact of this variance on the results. Therefore, if a proposed training method is accepted and comes to be widely used, it may reduce some amount of undesirable variance in the data. In this article, we focus on the training method for stationing in the MRI scanner. MRI scanning is expensive, and the proposed method we developed has the purpose of being cost-effective and thus generalizable to trainers around the world without regular access to an MRI scanner for training.
The method consists of two major components: training and testing. Training consists of two phases. Phase one is training the dog to chin target (i.e., station) in an open environment and phase two is training the dog to station in a mock MRI. Desensitization to the MRI occurs throughout the training phases, during separate, dedicated Auditory Exposure sessions. Testing consists of stationing in a portable mock MRI, in five different testing locations. The utility of this testing phase is to generalize the stationing behavior, facilitating transfer to the real MRI environment. The overall protocol is summarized in Figure 1.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/60192/60192fig1a.jpg" /> 
Figure 1: Protocol timeline. The protocol timeline is divided into two components, Training and Testing. Training is further divided into two phases, Open Environment and Mock MRI. Separate Auditory Exposure sessions occur during training as well. Testing consists of stationing in a portable mock MRI, in five different transfer locations (T1-T5). Once the dog has generalized the stationing behavior to criterion in five distinct transfer locations, the dog is ready for data collection in the real MRI environment. <a href="https://www.jove.com/files/ftp_upload/60192/60192fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
Depending on the phase, training and testing takes 25 to 75 minutes per week, per dog: one 10-minute Auditory Exposure session and two or more 5 to 30-minute Stationing sessions. This protocol can be completed in 25 weeks. During transfer testing, dogs execute several bouts of a 5-minute motionless down/stay and chin rest in a portable mock MRI (bore, radiofrequency coil, 90+ dB audio, ear padding) in five dissimilar locations. Transfer sessions occur once per week for 30-60 minutes, over five consecutive weeks. During MRI testing, dogs execute several bouts of the final stationing behavior during a 60-minute session of structural and functional data acquisition in a real MRI scanner.
Throughout training and testing, a chin rest is the behavior of focus. A chin rest is the dog touching his chin to an object&#39;s surface following some cue to target (i.e., rest his chin) to that surface. That cue to target can be physical (e.g., gesture, lure), verbal (e.g., spoken word &#34;rest&#34;), or an object (e.g., access to the chin rest itself). Fluent performance of the chin targeting behavior is critical to limiting head motion. In this protocol, the chin rest behavior is conditioned, maintained, and generalized to occur in multiple contexts (different rest apparatuses, in multiple locations) with increasing target duration (up to five minutes). Additionally, the trainer conditions and maintains strong performance of behaviors down and stay, as well as good stimulus control over the release cue &#34;Okay,&#34; the conditioned reinforcer and behavioral event marker &#34;click,&#34; and the Keep Going Signal (KGS) &#34;good&#34;8. Over the course of the protocol, multiple stimuli and apparatuses are introduced at specific stages and for specific intervals. These materials are easily and inexpensively procured. For full details, see the Table of Materials.

<table><tbody><tr><td>Acrylic Mock Radiofrequency Coil</td><td>Menards</td><td>TU59018594</td><td>Mock Radiofrequency (RF) Coil: 8&quot; diameter x 4&#039; Concrete Form Tube. Makes four mock RF coils; cut form tube in four even lengths for four 8&quot; diameter x 1&#039; mock RF coils.</td></tr><tr><td>Agility Tunnel</td><td>J&amp;amp;J Dog Supplies</td><td>TT053</td><td>Open Agility Training Tunnel</td></tr><tr><td>Bluetooth Speaker</td><td>Sharkk</td><td>SP-SK896WTR-GRY</td><td>Portable Scan Audio Playback: Waterproof Bluetooth Speaker Sharkk 2O IP67 Bluetooth Speaker Outdoor Pool Beach and Shower Portable Wireless Speaker</td></tr><tr><td>Cardboard Concrete Form Tube</td><td>Menards</td><td>TU10120014</td><td>Stationary Mock MRI Bore: Sonotube 24&quot; diameter x 12&#039; Standard Wall Water-Resistant Concrete Form. Makes two mock bores; cut form tube in half for two 24&quot; diameter x 6&#039; bores.</td></tr><tr><td>Chuckit Ball</td><td>Chuckit!</td><td>17030</td><td>Toy Reward: Chuckit! Ultra Ball</td></tr><tr><td>Decibel X</td><td>Skypaw</td><td></td><td>Decibel meter phone app</td></tr><tr><td>Exercise Mat</td><td></td><td></td><td>Foam chin rest: cut mat in half lengthwise. Roll up, and secure roll with hot glue. Cut chin-size notch in center with X-ACTO knife. Hot-glue velcro to bottom surface.</td></tr><tr><td>Folding Table</td><td></td><td></td><td>3&#039; x 6&#039; folding table</td></tr><tr><td>Microfiber Car Wax Applicator Pad</td><td>Viking Car Care</td><td>862400</td><td>Viking Car Care Microfiber Applicator Pads</td></tr><tr><td>Natural Balance Treat Log</td><td>Natual Balance</td><td>236020</td><td>Food Reward: E.g., Chicken Formula Dog Food Roll, 3.5-lb roll</td></tr><tr><td>Plywood</td><td></td><td></td><td>Platform: 2&quot;x4&quot;x6&#039; length of wood affixed to 3&#039;x6&#039; plywood board. Hot glue exercise mat on plywood board for traction. Braces: 3 4x4x4&quot; cubes cut at 45-degree angle affixed to ends of 1&quot;x4&quot;x3&#039; lengths of wood. Makes 3 braces.</td></tr><tr><td>Sand Bags</td><td>J&amp;amp;J Dog Supplies</td><td>AG155</td><td>J&amp;amp;J Professional Quality Sandbags x 2</td></tr><tr><td>Speaker System</td><td>Pioneer Electrics</td><td>HTD645DV</td><td>&lt;a href=&quot;https://www.pioneerelectronics.com/pio/pe/images/portal/cit_11221/93327312Operating%20Instructions%20S-HTD630.pdf&quot; target=&quot;_blank&quot;&gt;Stationary Scan Audio Playback: Pioneer HTD645DV 5 Disk DVD Home Theater System with Wireless Surround Speakers. Operating Instructions.&lt;/a&gt;</td></tr><tr><td>Towel</td><td></td><td></td><td>standard towel</td></tr></tbody></table>

training dogs for awake, unrestrained functional magnetic resonance imaging

We present a canine functional Magnetic Resonance Imaging (fMRI) training protocol that can be done in a cost-effective manner, with high-energy dogs, for acquisition of functional and structural data. This method of training dogs for awake, unrestrained fMRI employs a generalization procedure of stationing in several dissimilar locations to facilitate transfer of the stationing behavior to the real MRI scan environment; it does so without the need for extensive training time in the MRI scan environment, which can be expensive. Further, this method splits the training of a stationing (i.e., chin rest) behavior from desensitization to the MRI environment (i.e., 100+ decibel scan audio), the latter accomplished during dedicated Auditory Exposure conditioning sessions. The complete training and testing protocol required 14 hours and resulted in immediate transfer to novel locations. We also present examples of canine fMRI data that have been acquired from visual face processing and olfactory discrimination paradigms.

The mean number of repetitions of each session level is listed in Table 1. The complete training and testing protocol required 14 h (M = 13.55 h, range 12-16 h) and consisted of 90 sessions (range 87-93 sessions). Open environment training lasted 4.38 h (range 3-5 h), mock MRI training lasted 5.4 h (range 4.2-6.5 h), and transfer was 2.5 h divided into five 30 min sessions. Maintenance sessions at level 19 were conducted during transfer and are reflected in the complete t...

Watch this Scientific Journal Video about Training Dogs for Awake, Unrestrained Functional Magnetic Resonance Imaging at JoVE.com

Training Dogs for Awake, Unrestrained Functional Magnetic Resonance Imaging

Magnetic resonance imaging (MRI) on unrestrained awake dogs is a new method with several advantages over imaging with physical or chemical restraint. This protocol introduces a cost-effective training method that minimizes training in the MRI environment, which can be expensive, and maximizes the subject pool available for canine functional MRI.

We present a canine functional Magnetic Resonance Imaging (fMRI) training protocol that can be done in a cost-effective manner, with high-energy dogs, for ...

training-dogs-for-awake-unrestrained-functional-magnetic-resonance

Research

JoVE Journal

Behavior

7.5K Views.  Auburn University. Magnetic resonance imaging (MRI) on unrestrained awake dogs is a new method with several advantages over imaging with physical or chemical restraint. This protocol introduces a cost-effective training method that minimizes training in the MRI environment, which can be expensive, and maximizes the subject pool available for canine functional MRI.

Video: Training Dogs for Awake, Unrestrained Functional Magnetic Resonance Imaging

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful rest."1,2,3 It is thought the default mode network corresponds to self-referential or "internal mentation".2,3
It has been hypothesized that, in humans, activity within the default mode network is correlated with certain pathologies (for instance, hyper-activation has been linked to schizophrenia 4,5,6 and autism spectrum disorders 7 whilst hypo-activation of the network has been linked to Alzheimer's and other neurodegenerative diseases 8). As such, noninvasive modulation of this network may represent a potential therapeutic intervention for a number of neurological and psychiatric pathologies linked to abnormal network activation. One possible tool to effect this modulation is Transcranial Magnetic Stimulation: a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability (either increasing or decreasing it) via the application of localized magnetic field pulses.9
In order to explore the default mode network's propensity towards and tolerance of modulation, we will be combining TMS (to the left inferior parietal lobe) with functional magnetic resonance imaging (fMRI). Through this article, we will examine the protocol and considerations necessary to successfully combine these two neuroscientific tools.

In this article, we examine the methodology and considerations relevant to the combination of TMS and fMRI to examine the effects of brain stimulation on the default network.

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

Neuroscience

Functional Imaging of Auditory Cortex in Adult Cats using High-field fMRI

Current knowledge of sensory processing in the mammalian auditory system is mainly derived from electrophysiological studies in a variety of animal models, including monkeys, ferrets, bats, rodents, and cats. In order to draw suitable parallels between human and animal models of auditory function, it is important to establish a bridge between human functional imaging studies and animal electrophysiological studies. Functional magnetic resonance imaging (fMRI) is an established, minimally invasive method of measuring broad patterns of hemodynamic activity across different regions of the cerebral cortex. This technique is widely used to probe sensory function in the human brain, is a useful tool in linking studies of auditory processing in both humans and animals and has been successfully used to investigate auditory function in monkeys and rodents. The following protocol describes an experimental procedure for investigating auditory function in anesthetized adult cats by measuring stimulus-evoked hemodynamic changes in auditory cortex using fMRI. This method facilitates comparison of the hemodynamic responses across different models of auditory function thus leading to a better understanding of species-independent features of the mammalian auditory cortex.

Functional studies of the auditory system in mammals have traditionally been conducted using spatially-focused techniques such as electrophysiological recordings. The following protocol describes a method of visualizing large-scale patterns of evoked hemodynamic activity in the cat auditory cortex using functional magnetic resonance imaging.

Current knowledge of sensory processing in the mammalian auditory system is mainly derived from electrophysiological studies in a variety of animal ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Optogenetic Functional MRI

The investigation of the functional connectivity of precise neural circuits across the entire intact brain can be achieved through optogenetic functional magnetic resonance imaging (ofMRI), which is a novel technique that combines the relatively high spatial resolution of high-field fMRI with the precision of optogenetic stimulation. Fiber optics that enable delivery of specific wavelengths of light deep into the brain in vivo are implanted into regions of interest in order to specifically stimulate targeted cell types that have been genetically induced to express light-sensitive trans-membrane conductance channels, called opsins. fMRI is used to provide a non-invasive method of determining the brain's global dynamic response to optogenetic stimulation of specific neural circuits through measurement of the blood-oxygen-level-dependent (BOLD) signal, which provides an indirect measurement of neuronal activity. This protocol describes the construction of fiber optic implants, the implantation surgeries, the imaging with photostimulation and the data analysis required to successfully perform ofMRI. In summary, the precise stimulation and whole-brain monitoring ability of ofMRI are crucial factors in making ofMRI a powerful tool for the study of the connectomics of the brain in both healthy and diseased states.

This protocol describes the steps and data analysis required to successfully perform optogenetic functional magnetic resonance imaging (ofMRI). ofMRI is a novel technique that combines high-field fMRI readout with optogenetic stimulation, allowing for cell type-specific mapping of functional neural circuits and their dynamics across the whole living brain.

The investigation of the functional connectivity of precise neural circuits across the entire intact brain can be achieved through optogenetic functional ...

A Protocol for the Administration of Real-Time fMRI Neurofeedback Training

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control neuroplastic processes and correct abnormal function, or even shift functions from damaged tissue to physiologically healthy brain regions, hold the potential to dramatically improve overall health. Of the current neuroplastic interventions in development, neurofeedback training (NFT) from functional Magnetic Resonance Imaging (fMRI) has the advantages of being completely non-invasive, non-pharmacologic, and spatially localized to target brain regions, as well as having no known side effects. Furthermore, NFT techniques, initially developed using fMRI, can often be translated to exercises that can be performed outside of the scanner without the aid of medical professionals or sophisticated medical equipment. In fMRI NFT, the fMRI signal is measured from specific regions of the brain, processed, and presented to the participant in real-time. Through training, self-directed mental processing techniques, that regulate this signal and its underlying neurophysiologic correlates, are developed. FMRI NFT has been used to train volitional control over a wide range of brain regions with implications for several different cognitive, behavioral, and motor systems. Additionally, fMRI NFT has shown promise in a broad range of applications such as the treatment of neurologic disorders and the augmentation of baseline human performance. In this article, we present an fMRI NFT protocol developed at our institution for modulation of both healthy and abnormal brain function, as well as examples of using the method to target both cognitive and auditory regions of the brain.

The ability to induce and/or control neural plasticity may be critical in future treatments for neurologic disorders and the recovery from brain injury. In this paper, we present a protocol on the use of neurofeedback training with functional magnetic resonance imaging to modulate human brain function.

Neurologic disorders are characterized by abnormal cellular-, molecular-, and circuit-level functions in the brain. New methods to induce and control ...

Functional Magnetic Resonance Spectroscopy at 7 T in the Rat Barrel Cortex During Whisker Activation

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to technological developments over the last decade and the increase in magnetic field strength, it is now possible to obtain good resolution spectra in vivo in the rat brain. Neuroenergetics (i.e., the study of brain metabolism) and, especially, metabolic interactions between the different cell types have attracted more and more interest in recent years. Among these metabolic interactions, the existence of a lactate shuttle between neurons and astrocytes is still debated. It is, thus, of great interest to perform functional proton magnetic resonance spectroscopy (1H-MRS) in a rat model of brain activation and monitor lactate. However, the methyl lactate peak overlaps lipid resonance peaks and is difficult to quantify. The protocol described below allows metabolic and lactate fluctuations to be monitored in an activated brain area. Cerebral activation is obtained by whisker stimulation and 1H-MRS is performed in the corresponding activated barrel cortex, whose area is detected using blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI). All steps are fully described: the choice of anesthetics, coils, and sequences, achieving efficient whisker stimulation directly in the magnet, and data processing.

After checking by blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI) that the corresponding somatosensory barrel field cortex area (called S1BF) is correctly activated, the main goal of this study is to quantify lactate content fluctuations in the activated rat brains by localized proton magnetic resonance spectroscopy (1H-MRS) at 7 T.

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to ...

Real-Time fMRI Brain Mapping in Animals

Dynamic fMRI responses vary largely according to the physiological conditions of animals either under anesthesia or in awake states. We developed a real-time fMRI platform to guide experimenters to monitor fMRI responses instantaneously during acquisition, which can be used to modify the physiology of animals to achieve desired hemodynamic responses in animal brains. The real-time fMRI set-up is based on a 14.1T preclinical MRI system, enabling the real-time mapping of dynamic fMRI responses in the primary forepaw somatosensory cortex (FP-S1) of anesthetized rats. Instead of a retrospective analysis to investigate confounding sources leading to the variability of fMRI signals, the real-time fMRI platform provides a more effective scheme to identify dynamic fMRI responses using customized macro-functions and a common neuroimage analysis software in the MRI system. Also, it provides immediate troubleshooting feasibility and a real-time biofeedback stimulation paradigm for brain functional studies in animals.

Animal brain functional mapping can benefit from the real-time functional magnetic resonance imaging (fMRI) experimental set-up. Using the latest software implemented in the animal MRI system, we established a real-time monitoring platform for small animal fMRI.

Dynamic fMRI responses vary largely according to the physiological conditions of animals either under anesthesia or in awake states. We developed a ...

Acquisition of Resting-State Functional Magnetic Resonance Imaging Data in the Rat

Resting-state functional magnetic resonance imaging (rs-fMRI) has become an increasingly popular method to study brain function in a resting, non-task state. This protocol describes a preclinical survival method for obtaining rs-fMRI data. Combining low dose isoflurane with continuous infusion of the &#945;2 adrenergic receptor agonist dexmedetomidine provides a robust option for stable, high-quality data acquisition while preserving brain network function. Furthermore, this procedure allows for spontaneous breathing and near-normal physiology in the rat. Additional imaging sequences can be combined with resting-state acquisition creating experimental protocols with anesthetic stability of up to 5 h using this method. This protocol describes the setup of equipment, monitoring of rat physiology during four distinct phases of anesthesia, acquisition of resting-state scans, quality assessment of data, recovery of the animal, and a brief discussion of post-processing data analysis. This protocol can be used across a wide variety of preclinical rodent models to help reveal the resulting brain network changes that occur at rest.

This protocol describes a method for obtaining stable resting-state functional magnetic resonance imaging (rs-fMRI) data from a rat using low dose isoflurane in combination with low dose dexmedetomidine.

Resting-state functional magnetic resonance imaging (rs-fMRI) has become an increasingly popular method to study brain function in a resting, non-task ...

Head Implants for the Neuroimaging of Awake, Head-Fixed Rats

Anesthetics, commonly used in preclinical and fundamental scientific research, have a depressive influence on the metabolic, neuronal, and vascular functions of the brain and can adversely influence neurophysiological results. The use of awake animals for research studies is advantageous but poses the major challenge of keeping the animals calm and stationary to minimize motion artifacts throughout data acquisition. Awake imaging in smaller-sized rodents (e.g., mice) is very common but remains scant in rats as rats are bigger, stronger, and have a greater tendency to oppose movement restraints and head fixation over the long durations required for imaging. A new model of neuroimaging of awake, head-fixed rats using customized hand-sewn slings, 3D-printed head implants, head caps, and a headframe is described. The results acquired following a single trial of single-whisker stimulation suggest an increase in the intensity of the evoked functional response. The acquisition of the evoked functional response from awake, head-fixed rats is faster than that from anesthetized rats, reliable, reproducible, and can be used for repeated longitudinal studies.

A detailed new procedure for functional imaging of awake, head-fixed rats is described.

Anesthetics, commonly used in preclinical and fundamental scientific research, have a depressive influence on the metabolic, neuronal, and vascular ...

Through a Dog's Eyes: fMRI Decoding of Naturalistic Videos from the Dog Cortex

Recent advancements using machine learning and functional magnetic resonance imaging (fMRI) to decode visual stimuli from the human and nonhuman cortex have resulted in new insights into the nature of perception. However, this approach has yet to be applied substantially to animals other than primates, raising questions about the nature of such representations across the animal kingdom. Here, we used awake fMRI in two domestic dogs and two humans, obtained while each watched specially created dog-appropriate naturalistic videos. We then trained a neural net (Ivis) to classify the video content from a total of 90 min of recorded brain activity from each. We tested both an object-based classifier, attempting to discriminate categories such as dog, human, and car, and an action-based classifier, attempting to discriminate categories such as eating, sniffing, and talking. Compared to the two human subjects, for whom both types of classifier performed well above chance, only action-based classifiers were successful in decoding video content from the dogs. These results demonstrate the first known application of machine learning to decode naturalistic videos from the brain of a carnivore and suggest that the dog's-eye view of the world may be quite different from our own.

Machine learning algorithms have been trained to use patterns of brain activity to "decode" stimuli presented to humans. Here, we demonstrate that the same technique can decode naturalistic video content from the brains of two domestic dogs. We find that decoders based on the actions in the videos were successful in dogs.

Recent advancements using machine learning and functional magnetic resonance imaging (fMRI) to decode visual stimuli from the human and nonhuman cortex ...