Name: touchscreen sustained attention task (sat) for rats
Uploaded: 2017-09-15T15:00:00
Description: Watch this Scientific Journal Video about Touchscreen Sustained Attention Task SAT for Rats at JoVE.com

We would like to thank Joy Bergmann, Attilio Ceretti, Sarah Cohen, Nina Duncan, Arron Hall, Hanna Lefebo, and Madeleine Salvatore for their assistance with the behavior, and Dr. Vinay Parikh, Dr. Brittney Yegla, and Rob Cole for their advice on the SAT parameters. We would like to thank Dr. Jill McGaughy for tips on the training procedure. This work was supported by the National Institutes of Health grant NIMH 092438 and National Science Foundation grant IOS 1552416 to D.A.B.

All experiments were conducted in accordance with the National Institutes of Health guidelines and were approved by the Temple University Institutional Animal Use and Care Committee. This protocol was developed with adult (~60 days of age when starting the study) male and female Sprague-Dawley rats (Charles River).
1. Materials for Touchscreen SAT
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	<li>Ensure that a sound attenuation cubicle surrounds each touchscreen chamber and that chambers are equipped with a house light, tone gener...

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	<li>Cornblatt, B. A., Keilp, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+attention,+genetics,+and+the+pathophysiology+of+schizophrenia.">Impaired attention, genetics, and the pathophysiology of schizophrenia.</a> Schizphr Bull. 20 (1), 31-46 (1994).</li><li>Gmehlin, 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=Attentional+Lapses+of+Adults+with+Attention+Deficit+Hyperactivity+Disorder+in+Tasks+of+Sustained+Attention.">Attentional Lapses of Adults with Attention Deficit Hyperactivity Disorder in Tasks of Sustained Attention.</a> Arch Clin Neuropsychol. 31 (4), 343-357 (2016).</li><li>Perry, R. J., Hodges, J. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Failures+of+sustained+attention+in+life,+lab,+and+brain:+Ecological+validity+of+the+SART.">Failures of sustained attention in life, lab, and brain: Ecological validity of the SART.</a> Neuropsychologia. 48 (9), 2564-2570 (2010).</li><li>Becker, J. B., Prendergast, B. J., Liang, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Female+rats+are+not+more+variable+than+male+rats:+a+meta-analysis+of+neuroscience+studies.">Female rats are not more variable than male rats: a meta-analysis of neuroscience studies.</a> Biol Sex Differ. 7 (1), 34 (2016).</li><li>Cope, Z. A., Young, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Five-Choice+Continuous+Performance+Task+(5C-CPT):+A+Cross-Species+Relevant+Paradigm+for+Assessment+of+Vigilance+and+Response+Inhibition+in+Rodents.">The Five-Choice Continuous Performance Task (5C-CPT): A Cross-Species Relevant Paradigm for Assessment of Vigilance and Response Inhibition in Rodents.</a> Curr Protoc Neurosci. 4, 9-56 (2017).</li><li>Kim, C. 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=The+continuous+performance+test+(rCPT)+for+mice:+a+novel+operant+touchscreen+test+of+attentional+function.">The continuous performance test (rCPT) for mice: a novel operant touchscreen test of attentional function.</a> Psychopharmacology (Berl). 232 (21-22), 3947-3966 (2015).</li><li>Robbins, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+5-choice+serial+reaction+time+task:+behavioural+pharmacology+and+functional+neurochemistry.">The 5-choice serial reaction time task: behavioural pharmacology and functional neurochemistry.</a> Psychopharmacology (Berl). 163 (3-4), 362-380 (2002).</li><li>McGaughy, J., Sarter, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+vigilance+in+rats:+task+validation+and+effects+of+age,+amphetamine,+and+benzodiazepine+receptor+ligands.">Behavioral vigilance in rats: task validation and effects of age, amphetamine, and benzodiazepine receptor ligands.</a> Psychopharmacology (Berl). 117 (3), 340-357 (1995).</li><li>McGaughy, J., Sarter, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+ovariectomy,+192+IgG-saporin-induced+cortical+cholinergic+deafferentation,+and+administration+of+estradiol+on+sustained+attention+performance+in+rats.">Effects of ovariectomy, 192 IgG-saporin-induced cortical cholinergic deafferentation, and administration of estradiol on sustained attention performance in rats.</a> Behav neurosci. 113 (6), 1216-1232 (1999).</li><li>St Peters, M., Cherian, A. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Corticotropin+releasing+factor+impairs+sustained+attention+in+male+and+female+rats.">Corticotropin releasing factor impairs sustained attention in male and female rats.</a> Behav Brain Res. 296, 30-34 (2016).</li><li>Wicks, 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=Method+for+testing+sustained+attention+in+touchscreen+operant+chambers+in+rats.">Method for testing sustained attention in touchscreen operant chambers in rats.</a> J Neurosci Meth. 277, 30-37 (2017).</li><li>Hvoslef-Eide, M., Nilsson, S. R., Saksida, L. M., Bussey, T. 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C., 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=The+touchscreen+operant+platform+for+assessing+executive+function+in+rats+and+mice.">The touchscreen operant platform for assessing executive function in rats and mice.</a> Nat Protoc. 8 (10), 1985-2005 (2013).</li><li>Gill, T. M., Sarter, M., Givens, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+visual+attention+performance-associated+prefrontal+neuronal+activity:+evidence+for+cholinergic+modulation.">Sustained visual attention performance-associated prefrontal neuronal activity: evidence for cholinergic modulation.</a> J Neurosci. 20 (12), 4745-4757 (2000).</li><li>Mazure, C. M., Jones, D. 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Deficits in sustained attention are reported in neuropsychiatric, neurodevelopmental, and neurodegenerative disorders1,2,3, but the processes that contribute to optimal versus disrupted attention are understudied. This is, in part, because specialized equipment limits the easy adaptation of rodent attention tasks. However, touchscreen chambers are addressing this issue by providing the flexibility to run a variety of cognitive t...

Disorders ranging from attention deficit hyperactivity disorder (ADHD) to schizophrenia to Alzheimer&#39;s disease, share attentional impairments as a feature1,2,3. Deficits in sustained attention&#8212;the ability to monitor continuously a situation for intermittent and unpredictable events&#8212;are particularly disruptive, because sustained attention is crucial for selective and divided attention, as well as other cognitive processes4,5. Even in healthy people, difficulty with sustaining attention negatively affects cognition, impairing daily function6. Thus, understanding the neurobiological basis of sustained attention and how it becomes dysregulated could lead to interventions to improve cognition that would benefit many people.
In order to delineate circuits and molecular processes that contribute to proper versus disrupted attention, many researchers have turned to non-human animal models, where the manipulation of specific cell populations and molecular processes during attention tasks is possible. The effort has led to the development of a variety of operant attention tasks that can assess the ability to sustain attention7,8,9,10. One such paradigm, developed by McGaughy and Sarter (1995), is known as the sustained attention task (SAT). Rodents trained in SAT must distinguish signal trials, in which a signal light briefly flashes, from non-signal trials. Attentional demands can be increased by introducing a visual distractor (flashing houselight)10. The parameters critical for assessing attention in SAT are well documented and this task has been validated in male and female rats and mice10,11,12. A version has even been adapted for humans, highlighting the translational utility of SAT13. Importantly, the use of this task helped implicate the basal forebrain corticopetal system in sustained attention4. Specifically, cholinergic neurons in the nucleus basalis of Meynert (NBM)/ substantia inominata (SI) region of the basal forebrain that project to the prefrontal cortex are critical for hits, which are accurate responses on signal trials14,15. In contrast, GABAergic neurons in this region are thought to mediate performance during correct rejections (CRs), which are accurate responses on non-signal trials16. Once the basic circuit for this task was established, factors that modulate this circuit to impair attention ranging from stress hormones to neurotoxic proteins have been identified17,18. Collectively, these studies highlight the utility of SAT.
One limitation to implementing SAT in the laboratory is that the original procedure requires operant chambers permanently configured for the task, with a center panel light and two extendable levers, one designated for hits and one for CRs10. On a signal trial, the panel light briefly illuminates and then levers extend to indicate a 4 s response window. During that window, if the designated hit lever is pressed, then the rat receives a reward (either food or water). Incorrect lever presses on signal trials, known as misses, are not rewarded. On non-signaled trials, the panel light remains off, then levers are extended. While a CR is rewarded, incorrect lever presses on non-signaled trials, known as false alarms, are not rewarded. A failure to make any response during the response window counts as an omission. To expand the availability of SAT, we recently modified the task for touchscreen operant chambers, in which one wall is a touchscreen that can both display visual stimuli and record nose-poke responses19. Touchscreen chambers are becoming popular because of their versatility to run a variety of tasks (e.g., paired associate learning, reversal learning, etc.) with one piece of equipment20. Similar to other touchscreen procedures20,21,22, our adaptation of SAT requires the use of an opaque plastic touchscreen cover, called a mask, with holes cut into it. A central, circular hole allows for the presentation of a signal by presenting a white stimulus on the screen behind it, and two square response areas, one designated for hits and one for CRs, allow the rat to make touch responses19. In the traditional chambers, the 4-s response window is typically indicated with the presentation of the levers, but a previous study demonstrated that a tone can also be used to signal the response window23. In touchscreen SAT, we similarly signal the response window with a brief tone. Other training parameters (e.g., number of trials, intertrial interval, etc.) are kept similar between traditional and touchscreen SAT. We previously assessed performance between the traditional and touchscreen SAT versions and found that it was comparable, suggesting that touchscreen SAT is a valid way to measure sustained attention19. The touchscreen modification makes SAT more versatile because it can now be adapted for widespread use in touchscreen operant boxes.
The present protocol details how to run touchscreen SAT in the laboratory. The training schedules have been slightly altered from our previous report of the task19 to optimize acquisition time in female rats, which, unlike males, had difficulty advancing through the previous schedules24. Results illustrating typical performance in male and female rats are included, along with tips for troubleshooting issues with the task.

<table>
	<tbody>
		<tr>
			<td>Bussey-Saksida Rat Touch Screen Chambers Easy-Install System</td>
			<td>Lafayette Instrument</td>
			<td>80604-20</td>
			<td>Includes: Touch Screen, Chamber base with perforated floors, Large Feeder Reward Area (44.5mm wide x 92.5mm high), Speaker, Houselight, Tone Generator, SAC; Trapezoidal animal working area is 126mm wide at the Feeder, 240mm wide and 332 mm deep at the Screen, and 300 mm in height</td>
		</tr>
		<tr>
			<td>Sound Attenuation Cubicles (SAC)</td>
			<td>Lafayette Instrument</td>
			<td>80604-20</td>
			<td>Attenuation to approximately 35dB; 38kg in weight; External dimensions: 600mm wide x 670mm tall x 352mm deep; Internal dimensions: 540mm wide x 610mm tall x 532mm deep</td>
		</tr>
		<tr>
			<td>Abet II Software for Touch Screens</td>
			<td>Lafayette Instrument</td>
			<td>89505</td>
			<td>Version 2.19</td>
		</tr>
		<tr>
			<td>Whisker Multimedia</td>
			<td>Lafayette Instrument</td>
			<td>80698-1</td>
			<td>Required for Abet II Touch Operation</td>
		</tr>
		<tr>
			<td>Pellet Dispenser</td>
			<td>Lafayette Instrument</td>
			<td>80209-45</td>
			<td>45mg Interchangeable Pellet Size Wheel</td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>Lafayette Instrument</td>
			<td>80600-CAM</td>
			<td>Filtered and focused for IR light</td>
		</tr>
		<tr>
			<td>Microsoft Windows</td>
			<td>Microsoft</td>
			<td></td>
			<td>Windows 7-64 bit recommended</td>
		</tr>
		<tr>
			<td>Controller PC-Touch Screen Chambers</td>
			<td>Lafayette Instrument</td>
			<td>88530</td>
			<td>Dual Core Pentinum Processor; 2.5 GHz or greater; keyboard, mouse, and monitor; Installed PCI card, cable and expansion; Four RS232 ports; Four VGA ports</td>
		</tr>
		<tr>
			<td>Dustless Precision Pellets</td>
			<td>BioServe</td>
			<td>F0165</td>
			<td>45 mg</td>
		</tr>
		<tr>
			<td>Black Acrylic Mask</td>
			<td>Everything Plastic</td>
			<td>Custom Product</td>
			<td>Black polycarbonate with one matte finish side, 3.18 mm thick; Central circular opening, 28.58 mm diameter, 107.95 mm from the bottom, centered between the left and right side; Two square response areas, 28.57 mm x 28.57mm each positioned below and off center, one 85.725 mm from the left and one 85.725 mm from right side, and both 34.925 mm from the bottom.</td>
		</tr>
		<tr>
			<td>Utility Cart</td>
			<td>Fisher Scientific</td>
			<td>11-954-754</td>
			<td>30.75 in. x 18.38 in x 33 in</td>
		</tr>
		<tr>
			<td>Black towel</td>
			<td></td>
			<td></td>
			<td>Large enough to cover cages for animal transport</td>
		</tr>
		<tr>
			<td>Headlights with red lights</td>
			<td>Energizer</td>
			<td>HDL33A2E</td>
			<td>LED white and red light</td>
		</tr>
	</tbody>
</table>

touchscreen sustained attention task (sat) for rats

Sustained attention is the ability to monitor intermittent and unpredictable events over a prolonged period of time. This attentional process subserves other aspects of cognition and is disrupted in certain neurodevelopmental, neuropsychiatric, and neurodegenerative disorders. Thus, it is clinically important to identify mechanisms that impair and improve sustained attention. Such mechanisms are often first discovered using rodent models. Therefore, several behavior procedures for testing aspects of sustained attention have been developed for rodents. One, first described by McGaughy and Sarter (1995), called the sustained attention task (SAT), trains rats to distinguish between signal (i.e., brief light presentation) and non-signal trials. The signals are short and thus require careful attention to be perceived. Attentional demands can be increased further by introducing a distractor (e.g., flashing houselight). We have modified this task for touchscreen operant chambers, which are configured with a touchscreen on one wall that can present stimuli and record responses. Here we detail our protocol for SAT in touchscreen chambers. Additionally, we present standard measures of performance in male and female Sprague-Dawley rats. Comparable performance on this task in both sexes highlights its use for attention studies, especially as more researchers are including female rodents in their experimental design. Moreover, the easy implementation of SAT for the increasingly popular touchscreen chambers increases its utility.

For most attention studies, rats are trained to criteria on SAT and then manipulations to disrupt or improve attention are performed. Acquisition data is not typically presented, but we do so here to illustrate rates of acquisition in male and female Sprague-Dawley rats. To this end, we quantified the median number of days to reach criteria on SAT (starting on the Nose Poke Phase). Note that a few rats never made it to criteria (n = 5 males, and n = 1 female). The median days that it took...

Watch this Scientific Journal Video about Touchscreen Sustained Attention Task SAT for Rats at JoVE.com

Touchscreen Sustained Attention Task SAT for Rats

Sustained attention, or continuously monitoring situations for intermittent and unpredictable events, is a critical aspect of cognition. Here we detail how to test sustained attention in rats using touchscreen operant chambers. We demonstrate comparable performance in male and female rats, making this task useful for studying attention in both sexes.

Touchscreen Sustained Attention Task (SAT) for Rats

Sustained attention is the ability to monitor intermittent and unpredictable events over a prolonged period of time. This attentional process subserves ...

The overall goal of this behavioral task with touchscreen operant chambers is to measure the sustained attention capacity of rats. This method can help uncover the neuro biological mechanisms critical for appropriate sustained attention which is the monitoring of intermittent and unpredictable events over time. The main advantage of this technique is that we have adapted a well validated rodent sustained attention task to the touchscreen operant chamber.An apparatus of growing popularity with programmability to many cognitive tasks. The applications of this technique extend toward the study of neuropsychiatric and neurodegenerative disorders characterized by disrupted attention. Therapies for which, can be tested using the method presented here.Generally, individuals new to this method will struggle with setting up a system to ensure that the rats are being accurately tracked and that the data is being properly recorded. For this experiment, touchscreen chambers must be prepared within sound attenuation cubicles. Equip the chambers with the house light, a tone generator, a camera positioned above, a touchscreen and a pellet dispenser with the feeder reward area.The feeder reward area can be illuminated. Before loading the 45 milligram reward pellets, shake the dust off them using a flour sifter. This dust can clog the feeder.If the rats are outfitted with a head implant, use a feeder with a large aperture. Feed each chamber with a black acrylic mask to limit the response areas on the touchscreen. Leave a central circular hole for the light stimulus and two square response holes below the central hole positioned center.While in use, operate the chamber in a sound attenuation cubicle. This completes the chamber preparations. The text protocol describes how to set the perimeters for the experiment.For this experiment, rats can be housed individually or in groups. Prior to starting a study, handle the rats daily for five minutes per day, for five straight days. Always wear personal protective gear.Start the food restriction three days before starting the behavior shaping. On the first day, record each rats free-feeding weight and calculate their target weight which is 85%of the free-feeding weight. Prior to shaping, pre-expose the rats to 15 food reward pellets in their home cage.After the initial food restriction, adjust the food intake to maintain the target weight, allowing for slight weight gain over time. Prior to starting a new schedule, food restrict the rats a little more than usual, to increase their motivation. On days that rats do not perform the task, increase their food by at least one gram to compensate for not receiving pellet rewards.After each session, wait at least 30 minutes before providing food, to avoid making an association between the testing succession and their feeding time. If the rats are group housed, feed them separately for an hour before regrouping them. In preparation, power up the equipment.Ensure that the pellet dispensers are filled with pellets and ensure that the mask is in position in front of the touchscreen. Then, check the function of each box. Test the pellet dispenser, the screen, the lights and the tone generator.Testing the function of the chambers at the beginning of each day is critical because any one small malfunction can confuse the animals and prevent them from learning the task or performing at a normal level. Next, review the schedule for loading the boxes. Pay close attention to the test group and input the relevant information regarding animal ID, sex, experimental condition, user initials and other notes.With all the preparations made, retrieve the subjects. Generally, test the rats at least 30 minutes into their dark cycle. To work with the rats in the dark, use head mounted red lights.When transporting the rats during the dark cycle, cover their cages with black towels. Once in the behavior room, load the rats into their appropriate box and start the session for the sustained attention task. In the sustained attention task, rats are trained to distinguish signal trials from non-signal trials.Signal trials include a variable duration light flash followed by an audible tone and a four second response window. During the four seconds, if the rat then posed the correct area on the screen. In this case, it is the right response area, the rat is rewarded with a food pellet and this is called, a hit.If the rat pokes the other response area, this is considered a miss and the rat is not rewarded. A non-signal trial does not include the light and the rat must poke a different part of the screen to indicate that no signal was detected to earn a reward. A correct response on a non-signal trial is called, a correct rejection.If the rat pokes the other response area on a non-signal trial, this is considered a false alarm and is not rewarded. The correct response areas for signal and non-signal trials should be counter-balanced between subjects. For signal and non-signal trials, a failure to make any response during the response window counts as a, no mission and is not rewarded.There is also a distracter version of the sustained attention task, where the house light flashes during the second of three blocks of trials. After starting a session, monitor the rats during the first several minutes, to ensure that everything is functioning properly. Then, check their performance intermittently.Once a session is complete, record the summary data and any issues in the notebook. Between trials, wipe down each chamber with 10%ethanol and replace the bedding. After running all the trials for the day, thoroughly clean the chambers with 10%ethanol, making sure the floor grid is as clean as possible.Then, replace the bedding. After every session, it is a good idea to fully update the spreadsheets and back them up to a separate server. The median number of days to reach criteria on SAT was 54.5 days for male rats and 62 days for female rats with no significant difference.Performance measures on the SAT task showed that based on the percentages of hits and correct rejections, males and females were similarly accurate on the signaled and non-signaled trials. An overall measure of intentional performance, the vigilance index or VI, showed similar measures between the sexes as well. Nor were there sex differences in the low percentages of omissions recorded.During the tasks, the visual signal varied between 500, 50 and 25 milliseconds and performance declined with the shorter signals. There was no main effect based on sex. To increase attentional demands, the house light was flashed as a distracter during the second of the three trial blocks, in one subset of rats.As expected, there was a main effect for the trial block. With performance declining in block two and recovering in block three. Unexpectedly, females performed significantly worse than males.After watching this video, you should have a good understanding of how to test sustained attention in rats using touchscreen operant chambers. While attempting this procedure, it's important to remember to keep track of each individual animal's performance and adjust feeding as necessary to ensure that they have acquired the task as quickly as possible. Don't forget, that when working with rodents, protocol should first be approved by IACUC and personal protective equipment should always be worn.

touchscreen-sustained-attention-task-sat-for-rats

Research

JoVE Journal

Neuroscience

Correlating Behavioral Responses to fMRI Signals from Human Prefrontal Cortex: Examining Cognitive Processes Using Task Analysis

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar tasks. Participants' behavior during task performance in an fMRI scanner can then be correlated to the brain activity using the blood-oxygen-level-dependent signal. We measure behavior to be able to sort correct trials, where the subject performed the task correctly and then be able to examine the brain signals related to correct performance. Conversely, if subjects do not perform the task correctly, and these trials are included in the same analysis with the correct trials we would introduce trials that were not only for correct performance. Thus, in many cases these errors can be used themselves to then correlate brain activity to them. We describe two complementary tasks that are used in our lab to examine the brain during suppression of an automatic responses: the stroop1 and anti-saccade tasks. The emotional stroop paradigm instructs participants to either report the superimposed emotional 'word' across the affective faces or the facial 'expressions' of the face stimuli1,2. When the word and the facial expression refer to different emotions, a conflict between what must be said and what is automatically read occurs. The participant has to resolve the conflict between two simultaneously competing processes of word reading and facial expression. Our urge to read out a word leads to strong 'stimulus-response (SR)' associations; hence inhibiting these strong SR's is difficult and participants are prone to making errors. Overcoming this conflict and directing attention away from the face or the word requires the subject to inhibit bottom up processes which typically directs attention to the more salient stimulus. Similarly, in the anti-saccade task3,4,5,6, where an instruction cue is used to direct only attention to a peripheral stimulus location but then the eye movement is made to the mirror opposite position. Yet again we measure behavior by recording the eye movements of participants which allows for the sorting of the behavioral responses into correct and error trials7 which then can be correlated to brain activity. Neuroimaging now allows researchers to measure different behaviors of correct and error trials that are indicative of different cognitive processes and pinpoint the different neural networks involved.

The goal of our research is to correlate behavior to brain activity. Accurate behavioral measures and imaging techniques allow us to elucidate brain-behavior relationships.

The aim of this methods paper is to describe how to implement a neuroimaging technique to examine complementary brain processes engaged by two similar ...

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

Mapping Cortical Dynamics Using Simultaneous MEG/EEG and Anatomically-constrained Minimum-norm Estimates: an Auditory Attention Example

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the cortical networks involved in dynamical perceptual and cognitive tasks, such as attending to different sounds in a cocktail party. Many past studies have employed data recorded at the sensor level only, i.e., the magnetic fields or the electric potentials recorded outside and on the scalp, and have usually focused on activity that is time-locked to the stimulus presentation. This type of event-related field / potential analysis is particularly useful when there are only a small number of distinct dipolar patterns that can be isolated and identified in space and time. Alternatively, by utilizing anatomical information, these distinct field patterns can be localized as current sources on the cortex. However, for a more sustained response that may not be time-locked to a specific stimulus (e.g., in preparation for listening to one of the two simultaneously presented spoken digits based on the cued auditory feature) or may be distributed across multiple spatial locations unknown a priori, the recruitment of a distributed cortical network may not be adequately captured by using a limited number of focal sources.
Here, we describe a procedure that employs individual anatomical MRI data to establish a relationship between the sensor information and the dipole activation on the cortex through the use of minimum-norm estimates (MNE). This inverse imaging approach provides us a tool for distributed source analysis. For illustrative purposes, we will describe all procedures using FreeSurfer and MNE software, both freely available. We will summarize the MRI sequences and analysis steps required to produce a forward model that enables us to relate the expected field pattern caused by the dipoles distributed on the cortex onto the M/EEG sensors. Next, we will step through the necessary processes that facilitate us in denoising the sensor data from environmental and physiological contaminants. We will then outline the procedure for combining and mapping MEG/EEG sensor data onto the cortical space, thereby producing a family of time-series of cortical dipole activation on the brain surface (or "brain movies") related to each experimental condition. Finally, we will highlight a few statistical techniques that enable us to make scientific inference across a subject population (i.e., perform group-level analysis) based on a common cortical coordinate space.

We use magneto- and electroencephalography (MEG/EEG), combined with anatomical information captured by magnetic resonance imaging (MRI), to map the dynamics of the cortical network associated with auditory attention.

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the ...

Conducting Concurrent Electroencephalography and Functional Near-Infrared Spectroscopy Recordings with a Flanker Task

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by inspecting the relationship between the neural and hemodynamic signals. EEG is an electrophysiological technology that can measure the rapid neuronal activity of the cortex, whereas fNIRS relies on the hemodynamic responses to infer brain activation. The combination of EEG and fNIRS neuroimaging techniques can identify more features and reveal more information associated with the functioning of the brain. In this protocol, fused EEG-fNIRS measurements were performed for concurrent recordings of evoked-electrical potentials and hemodynamic responses during a Flanker task. In addition, the critical steps for setting up the hardware and software system as well as the procedures for data acquisition and analysis were provided and discussed in detail. It is expected that the present protocol can pave a new avenue for improving the understanding of the neural mechanisms underlying various cognitive processes by using the EEG and fNIRS signals.

The present protocol describes how to perform concurrent EEG and fNIRS recordings and how to inspect the relationship between the EEG and fNIRS data.&#160;

Concurrent EEG and fNIRS recordings offer an excellent opportunity to gain a full understanding of the neural mechanism of cognitive processing by ...

Event Related Potentials (ERPs) and other EEG Based Methods for Extracting Biomarkers of Brain Dysfunction: Examples from Pediatric Attention Deficit/Hyperactivity Disorder (ADHD)

Neuropsychiatric diagnoses like ADHD are based on subjective methods like interviews, rating scales and observations. There is a need for more brain-based supplements. Stimulant medication is the most common treatment for ADHD. Clinically useful predictors of response have so far not been reported. The aim of this paper is to describe the EEG based methods we apply to extract potential biomarkers for brain dysfunction. Examples relate to biomarkers for pediatric ADHD, and prediction of medication response. The main emphasis is on Event Related Potentials (ERPs).
A nineteen channel EEG is recorded during a 3 min eyes-opened task, a 3 min eyes-closed task, and a 20 min cued visual GO/NOGO task (VCPT). ERPs are recorded during this task. The goal of the ERP protocol is to extract biomarkers of assumed brain dysfunctions that significantly differentiate between a patient group and healthy controls. The protocol includes recording during standard conditions and artifact correction. ERP waves can be used or transformed into latent components. The components of the patient group are compared with controls, empathizing components that, when compared, show relatively high effect sizes. Sub-groups of the patients are selected on the basis of the cluster analysis in the space of the components. Treatment procedure (such as medication, tDCS or neurofeedback protocol) can be applied and the changes in components related to treatment in the subgroups are observed, forming the basis for clinical recommendations.
The methods described were applied in a study of 87 pediatric ADHD patients. The index of medication response discriminated significantly between responders and non-responders with a large, and clinically meaningful effect size (d = 1.84). In an ongoing study comparing ADHD children with matched controls, several variables discriminate significantly between patients and controls. The global index will exceed d = .8. The EEG based methods described here could be clinically meaningful.

Event Related Potentials ERPs and other EEG Based Methods for Extracting Biomarkers of Brain Dysfunction: Examples from Pediatric Attention Deficit/Hyperactivity Disorder ADHD

EEG-methods are applied for extracting biomarkers of brain dysfunctions. The focus is on multi-channel event-related potentials (ERPs) recorded in a cued GO/NOGO task. Non-brain artifacts are corrected and ERPs are compared with the normative data. Examples relate to biomarkers for ADHD diagnosis and prediction of medication response.

Neuropsychiatric diagnoses like ADHD are based on subjective methods like interviews, rating scales and observations. There is a need for more brain-based ...

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. Several recent studies have demonstrated the effectiveness of chronic VNS in enhancing central nervous system plasticity to improve motor rehabilitation, extinction learning, and sensory discrimination. Construction of chronically implantable devices for use in such studies is challenging due to rats&#8217; small size, and typical protocols require extensive training of personnel and time-consuming microfabrication methods. Alternatively, commercially available implantable cuff electrodes can be purchased at a significantly higher cost. In this protocol, we present a simple, low-cost method for construction of small, chronically implantable peripheral nerve cuff electrodes for use in rats. We validate the short and long-term reliability of our cuff electrodes by demonstrating that VNS in ketamine/xylazine anesthetized rats produces decreases in breathing rate consistent with activation of the Hering-Breuer reflex, both at the time of implantation and up to 10 weeks after device implantation. We further demonstrate the suitability of the cuff electrodes for use in chronic stimulation studies by pairing VNS with skilled lever press performance to induce motor cortical map plasticity.

Existing approaches for constructing chronically implantable peripheral nerve cuff electrodes for use in small rodents often require specialized equipment and/or highly trained personnel. In this protocol we demonstrate a simple, low-cost approach for fabricating chronically implantable cuff electrodes, and demonstrate their effectiveness for vagus nerve stimulation (VNS) in rats.

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...

The Impact of Motor Task Conditions on Goal-Directed Arm Reaching Kinematics and Trunk Compensation in Chronic Stroke Survivors

Trunk compensation is the most common movement strategy to substitute for upper extremity (UE) motor deficits in chronic stroke survivors. There is a lack of evidence examining how task conditions impact trunk compensation and goal-directed arm reaching kinematics. This protocol aims to investigate the impact of task conditions, including task difficulty and complexity, on goal-directed arm reaching kinematics. Two non-disabled young adults and two chronic stroke survivors with mild UE motor impairment were recruited for testing the protocol. Each participant performed goal-directed arm reaches with four different task conditions (2 task difficulties [large vs. small targets] X 2 task complexities [pointing vs. picking up]). The task goal was to reach and point at a target or pick up an object located 20 cm in front of the home position as quickly as possible with a stylus or a pair of chopsticks, respectively, in response to an auditory cue. Participants performed ten reaches per task condition. A 3-dimensional motion capture camera system was used to record trunk and arm kinematics. Representative results showed that there was a significant increase in movement duration, movement jerkiness, and trunk compensation as a function of task complexity, but not task difficulty in all participants. Chronic stroke survivors showed significantly slower, jerkier, and more feedback-dependent arm reaches and significantly more compensatory trunk movements than non-disabled adults. Our representative results support that this protocol can be used to investigate the impact of task conditions on motor control strategies in chronic stroke survivors with mild UE motor impairment.

This protocol is intended to investigate the impact of task conditions on movement strategies in chronic stroke survivors. Further, this protocol can be used to examine if a restriction in elbow extension induced by neuromuscular electrical stimulation causes trunk compensation during goal-directed arm reaches in non-disabled adults.

Trunk compensation is the most common movement strategy to substitute for upper extremity (UE) motor deficits in chronic stroke survivors. There is a lack ...

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 ...

Dual-Task Stroop Paradigm for Detecting Cognitive Deficits in High-Functioning Stroke Patients

General clinical cognitive assessment scales are not sensitive enough to cognitive impairment in high-functioning stroke patients. The dual-task assessment has advantages for identifying cognitive deficits in high-functioning stroke patients and has been gradually applied in clinical assessment and cognitive training. Moreover, the Stroop paradigm has higher sensitivity and specificity for attentional assessment than conventional clinical cognitive assessment scales. Therefore, this study presents the dual-task assessment based on the Stroop paradigm to identify cognitive deficits in high-functioning stroke patients. This study demonstrates a single- and dual-task evaluation based on the Stroop paradigm and confirms its feasibility through case experiments and synchronized functional near-infrared spectroscopy evaluation. The Stroop reaction time and correct rate are used as the main indicators to evaluate the cognitive level of the subjects. This study protocol aims to provide new ideas to figure out the ceiling effect in general clinical assessment failure for high-functioning stroke patients.

Clinical assessment scales are notsensitive enough to cognitive dysfunction in high-functioning stroke patients. The dual-task paradigm presents advantages and potential in the assessment and cognitive training of cognitive dysfunction. The study here proposes a dual-task Stroop paradigm to identify cognitive dysfunction in high-functioning stroke patients.

General clinical cognitive assessment scales are not sensitive enough to cognitive impairment in high-functioning stroke patients. The dual-task ...

Surgery and Behavioral Testing in the Tibial Neuroma Transposition Model in Rats

The tibial neuroma transposition (TNT) is a rat model in which allodynia at the neuroma site (tibial nerve) can be independently evaluated from allodynia at the plantar surface of the hind paw innervated by the intact sural nerve. This TNT model is suitable to test therapies for neuroma pain, such as the potential superiority of certain surgical therapies that are already used in the clinic, or to evaluate new drugs and their effect on both pain modalities in the same animal. In this model, a distal lesion (neurotmesis) is made in the tibial nerve, and the proximal nerve end is transposed and fixed subcutaneously and pretibially to enable assessments of the neuroma site with a 15 g Von Frey monofilament. To assess allodynia over the sural nerve, Von Frey monofilaments can be used via the up-down method on the plantar lateral region of the hind paw. After cutting the tibial nerve, mechanical hypersensitivity develops at the neuroma site within 1 week after surgery and persists at least until 12 weeks after surgery. Allodynia at the sural innervated plantar surface develops within 3 weeks after surgery compared to the contralateral limb. At 12 weeks, a neuroma forms on the proximal end of the severed tibial nerve, indicated by dispersion and swirling of axons. For the TNT model surgery, multiple critical (micro)surgical steps need to be followed, and some surgery practice under terminal anesthesia is advised. Compared to other neuropathic pain models, such as the spared nerve injury model, allodynia over the neuroma site can be independently tested from sural nerve hypersensitivity in the TNT model. However, the neuroma site can be tested only in rats, not in mice. The tips and directions provided in this protocol can help research groups working on pain successfully implement the TNT model in their facility.

This protocol describes the tibial neuroma transposition model, which entails a lesion of the tibial nerve with subsequent transposition of the proximal nerve end toward a subcutaneous pretibial or lateral position. Behavioral testing of neuroma pain and plantar hyperalgesia is quantified using Von Frey monofilaments.

The tibial neuroma transposition (TNT) is a rat model in which allodynia at the neuroma site (tibial nerve) can be independently evaluated from allodynia ...