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.

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