Aby wyświetlić tę treść, wymagana jest subskrypcja JoVE. Zaloguj się lub rozpocznij bezpłatny okres próbny.
Method Article
* Wspomniani autorzy wnieśli do projektu równy wkład.
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 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.
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; Querétaro, 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.
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. Please click here to view a larger version of this figure.
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'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 "rest"), 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 "Okay," the conditioned reinforcer and behavioral event marker "click," and the Keep Going Signal (KGS) "good"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.
Access restricted. Please log in or start a trial to view this content.
Ethical approval for these methods was obtained from the Auburn University Institutional Animal Care and Use Committee and all methods were performed in accordance with their guidelines and regulations. For auditory exposure, progression through the sessions is based on week number. For stationing sessions, a specific session-specified performance criterion (e.g., at least eleven-second duration of chin targeting), must be met before the trainer may advance the dog to the next session in that training phase. Otherwise, that step is repeated.
1. Auditory Exposure sessions
NOTE: These sessions constitute passive exposure and active classical counterconditioning of a positive Conditioned Emotional Response (CER) to MRI scanner noise; the scanner noise is established as a stimulus predicting the access to toy play or food rewards. Exposure sessions occur once per week for approximately 10 min.
Figure 2: Active Exposure. Active Exposure (AE) is a short-delay classical conditioning procedure. 10 s CS (i.e., scan audio presented by itself), 20 s CS + US (i.e., ball and scan audio presented together), 10 s delay (no ball, no scan audio). After this delay, the trial starts over. There are ten trials per session, with incremental volume increases over sessions. Please click here to view a larger version of this figure.
NOTE: Collect the following audio: MRI scanner baseline, shim, localizer (scout), MPRAGE, GRE Field, EPI, Multiband EPI, DTI, and RESOLVE DTI, using, for example, a smartphone's audio recording app through the open door of a 3T MRI suite during phantom scans. Determine volume level of audio playback during training sessions via a decibel meter phone app.
2. Stationing sessions
NOTE: These sessions are divided into two phases: Open environment and Mock MRI. After the chin-to-object target is learned, durations are increased on a percentile schedule of 10% increases. As new elements and pieces of equipment are added into the training context, certain criteria of the behavior (e.g., duration) are temporarily relaxed:
1) In the stationing sessions, the trainer trains a nose-touch behavior to a folded towel and then a chin rest on a folded towel. That chin rest behavior is generalized to occur in a foam chin rest and gradually built to a 5 min bout duration.
2) Simultaneously, robust down and stay behaviors are built and maintained.
3) Those behaviors are then conditioned to occur in an enclosed space (i.e., tunnel) and at a 3' elevation.
4) The dog is then acclimated to the head enclosure (mock human extremity RF coil).
5) Ear padding is introduced, and scan audio is (re)introduced in the context of the stationing behavior.
The dog will ultimately be able to perform a robust chin rest with head and body enclosed at a 3' elevation, with ear padding and scan audio playing at 90 + decibels (dB), for at least 5 min bouts. On reinforcement - some dogs are inherently more motivated by food, whereas others are more motivated by play or praise9. In "click-then-treat" (C/T), the T does not necessarily mean food treats, rather it refers to the reward procedure, whatever that may be for that particular dog at that particular stage in its training. Although food rewards lend themselves to higher rates and stiller repetitions of behavior, whatever the dog prefers can be used initially, even if it is high-motion play (e.g., ball, tug). As the chin target behavior becomes more resilient against distraction and duration, transition to using food rewards. Eventually, toy play can be saved for long-duration or chained bouts of chin rest performance.
3. Transfer
NOTE: 1) Upon reaching final criterion of the stationing behavior in the mock MRI training location (5 min down-stay and chin rest in mock bore and mock RF coil while wearing ear padding, with scanner noise playing at 80-110 dB), the dog undergoes five distinct location transfer (generalization) sessions. During these transfer sessions the dog stations to the above criteria in several indoor and outdoor locations that are as unique as possible, with different sights, sounds, and degrees of social distraction across settings (e.g., secluded grass field, quiet academic building hallway, busy academic building lobby, crowded bus stop, loud water treatment plant)8.
4. MRI
Access restricted. Please log in or start a trial to view this content.
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...
Access restricted. Please log in or start a trial to view this content.
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...
Access restricted. Please log in or start a trial to view this content.
The authors have nothing to disclose.
We are grateful to Canine Performance Sciences and Auburn University Departments of Psychology and Electrical & Computer Engineering. This work was supported by the Association of Professional Dog Trainers.
Access restricted. Please log in or start a trial to view this content.
Name | Company | Catalog Number | Comments |
Acrylic Mock Radiofrequency Coil | Menards | TU59018594 | Mock Radiofrequency (RF) Coil: 8" diameter x 4' Concrete Form Tube. Makes four mock RF coils; cut form tube in four even lengths for four 8" diameter x 1' mock RF coils. |
Agility Tunnel | J&J Dog Supplies | TT053 | Open Agility Training Tunnel |
Bluetooth Speaker | Sharkk | SP-SK896WTR-GRY | Portable Scan Audio Playback: Waterproof Bluetooth Speaker Sharkk 2O IP67 Bluetooth Speaker Outdoor Pool Beach and Shower Portable Wireless Speaker |
Cardboard Concrete Form Tube | Menards | TU10120014 | Stationary Mock MRI Bore: Sonotube 24" diameter x 12' Standard Wall Water-Resistant Concrete Form. Makes two mock bores; cut form tube in half for two 24" diameter x 6' bores. |
Chuckit Ball | Chuckit! | 17030 | Toy Reward: Chuckit! Ultra Ball |
Decibel X | Skypaw | Decibel meter phone app | |
Exercise Mat | 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. | ||
Folding Table | 3' x 6' folding table | ||
Microfiber Car Wax Applicator Pad | Viking Car Care | 862400 | Viking Car Care Microfiber Applicator Pads |
Natural Balance Treat Log | Natual Balance | 236020 | Food Reward: E.g., Chicken Formula Dog Food Roll, 3.5-lb roll |
Plywood | Platform: 2"x4"x6' length of wood affixed to 3'x6' plywood board. Hot glue exercise mat on plywood board for traction. Braces: 3 4x4x4" cubes cut at 45-degree angle affixed to ends of 1"x4"x3' lengths of wood. Makes 3 braces. | ||
Sand Bags | J&J Dog Supplies | AG155 | J&J Professional Quality Sandbags x 2 |
Speaker System | Pioneer Electrics | HTD645DV | Stationary Scan Audio Playback: Pioneer HTD645DV 5 Disk DVD Home Theater System with Wireless Surround Speakers. Operating Instructions. |
Towel | standard towel |
Access restricted. Please log in or start a trial to view this content.
Zapytaj o uprawnienia na użycie tekstu lub obrazów z tego artykułu JoVE
Zapytaj o uprawnieniaThis article has been published
Video Coming Soon
Copyright © 2025 MyJoVE Corporation. Wszelkie prawa zastrzeżone