Name: place and response learning in the open-field tower maze
Uploaded: 2015-10-28T15:00:00
Description: Watch this Scientific Journal Video about Place and Response Learning in the Open-field Tower Maze at JoVE.com

The authors would like to thank Christopher Newport University for funding the preparation of this manuscript and The University of Vermont for funding the experiments described in the representative results section.

This protocol is approved by the Institutional Animal Care and Use Committee of Christopher Newport University.
1. Construction of the OFTM Apparatus
<ol>
	<li>Construct the boundary wall (40 cm high and 183 cm diameter) from three pieces of white laminate. Construct the circular maze floor (183 cm diameter) from black laminate and glue it to a plastic table top. Secure the boundary wall to the edge of the table with screws and glue.</li>
	<li>Decorate the interior surface of the boundary wall: Leave one quarter...

<ol>
	<li>Luine, V., 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=Repeated+stress+causes+reversible+impairments+of+spatial+memory+performance.">Repeated stress causes reversible impairments of spatial memory performance.</a> Brain Res. 639 (1), 167-170 (1994).</li><li>Lipatova, O., 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=Effects+of+continuous+vs.+cycling+estrogen+replacement+on+the+acquisition,+retention+and+expression+of+place-+and+response-learning+in+the+open-field+tower+maze.">Effects of continuous vs. cycling estrogen replacement on the acquisition, retention and expression of place- and response-learning in the open-field tower maze.</a> Neurobiol Learn Mem. 114, 81-89 (2014).</li><li>Packard, M. G., McGaugh, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Double+dissociation+of+fornix+and+caudate+nucleus+lesions+on+acquisition+of+two+water+maze+tasks:+further+evidence+for+multiple+memory+systems.">Double dissociation of fornix and caudate nucleus lesions on acquisition of two water maze tasks: further evidence for multiple memory systems.</a> Behav Neurosci. 106 (3), 439-446 (1992).</li><li>Packard, M. G., McGaugh, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inactivation+of+hippocampus+or+caudate+nucleus+with+lidocaine+differentially+affects+expression+of+place+and+response+learning.">Inactivation of hippocampus or caudate nucleus with lidocaine differentially affects expression of place and response learning.</a> Neurobiol Learn Mem. 65 (1), 65-72 (1996).</li><li>Poldrack, R. A., Packard, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Competition+among+multiple+memory+systems:+converging+evidence+from+animal+and+human+brain+studies.">Competition among multiple memory systems: converging evidence from animal and human brain studies.</a> Neuropsychologia. 41 (3), 245-251 (2003).</li><li>Quinlan, M. G., Hussain, D., Brake, W. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+cognitive+strategies+in+rats:+the+role+of+estradiol+and+its+interaction+with+dopamine.">Use of cognitive strategies in rats: the role of estradiol and its interaction with dopamine.</a> Horm Behav. 53 (1), 185-191 (2008).</li><li>Korol, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+estrogen+in+balancing+contributions+from+multiple+memory+systems.">Role of estrogen in balancing contributions from multiple memory systems.</a> Neurobiol Learn Mem. 82, 309-323 (2004).</li><li>Cole, M. R., Clipperton, A., Walt, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Place+versus+response+learning+in+rats.">Place versus response learning in rats.</a> Learn Behav. 35 (4), 214-224 (2007).</li><li>Prut, L., Belzung, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+open+field+as+a+paradigm+to+measure+the+effects+of+drugs+on+anxiety-like+behaviors:+a+review.">The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: a review.</a> Eur J Pharmaco. 463 (1-3), 3-33 (2003).</li><li>Harrison, F. E., Hosseini, A. H., McDonald, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+anxiety+and+stress+responses+in+water+maze+and+Barnes+maze+spatial+memory+tasks.">Endogenous anxiety and stress responses in water maze and Barnes maze spatial memory tasks.</a> Behav Brain Res. 198 (1), 247-251 (2009).</li><li>Bowman, R. E., Beck, K. D., Luine, V. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+stress+effects+on+memory:+sex+differences+in+performance+and+monoaminergic+activity.">Chronic stress effects on memory: sex differences in performance and monoaminergic activity.</a> Horm Behav. 43 (1), 48-59 (2003).</li><li>Lunga, P., Herbert, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=17Beta-oestradiol+modulates+glucocorticoid,+neural+and+behavioural+adaptations+to+repeated+restraint+stress+in+female+rats.">17Beta-oestradiol modulates glucocorticoid, neural and behavioural adaptations to repeated restraint stress in female rats.</a> J Neuroendocrinol. 16 (9), 776-7785 (2004).</li><li>Newhouse, P. A., 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=Estrogen+treatment+impairs+cognitive+performance+after+psychosocial+stress+and+monoamine+depletion+in+postmenopausal+women.">Estrogen treatment impairs cognitive performance after psychosocial stress and monoamine depletion in postmenopausal women.</a> Menopause. 17 (4), 860-873 (2010).</li><li>Lipatova, O., Toufexis, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estrogen+enhances+the+retention+of+spatial+reference+memory+in+the+open+field+tower+task,+but+disrupts+the+expression+of+spatial+memory+following+a+novel+start+position.">Estrogen enhances the retention of spatial reference memory in the open field tower task, but disrupts the expression of spatial memory following a novel start position.</a> Neurobiol Learn Mem. 99, 50-58 (2012).</li><li>Lipatova, O., 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=Recency-to-primacy+shift+in+cue+competition.">Recency-to-primacy shift in cue competition.</a> J Exp Psychol Anim Behav Process. 32 (4), 396-406 (2006).</li><li>De La Casa, L. G., Timberlake, W. <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+preexposure+and+retention+interval+placement+on+latent+inhibition+and+perceptual+learning+in+a+choice-maze+discrimination+task.">Effects of preexposure and retention interval placement on latent inhibition and perceptual learning in a choice-maze discrimination task.</a> Learn Behav. 34 (2), 193-201 (2006).</li><li>Liang, K. 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=Buspirone+impaired+acquisition+and+retention+in+avoidance+tasks:+involvement+of+the+hippocampus.">Buspirone impaired acquisition and retention in avoidance tasks: involvement of the hippocampus.</a> Chin J Physiol. 41 (1), 33-44 (1998).</li><li>Gisquet-Verrier, P., Alexinsky, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+contextual+change+determine+long-term+forgetting.">Does contextual change determine long-term forgetting.</a> Animal Learning and Behavior. 14 (4), 349-358 (1986).</li><li>Gisquet-Verrier, 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=Exposure+to+retrieval+cues+improves+retention+performance+and+induces+changes+in+ACTH+and+corticosterone+release.">Exposure to retrieval cues improves retention performance and induces changes in ACTH and corticosterone release.</a> Psychoneuroendocrinology. 29 (4), 529-556 (2004).</li><li>Packard, M. G., Hirsh, R., White, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+effects+of+fornix+and+caudate+nucleus+lesions+on+two+radial+maze+tasks:+evidence+for+multiple+memory+systems.">Differential effects of fornix and caudate nucleus lesions on two radial maze tasks: evidence for multiple memory systems.</a> J Neurosci. 9 (5), 1465-1472 (1989).</li><li>Devan, B. D., McDonald, R. J., White, N. 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+medial+and+lateral+caudate-putamen+lesions+on+place-+and+cue-guided+behaviors+in+the+water+maze:+relation+to+thigmotaxis.">Effects of medial and lateral caudate-putamen lesions on place- and cue-guided behaviors in the water maze: relation to thigmotaxis.</a> Behav Brain Res. 100 (1-2), 5-14 (1999).</li></ol>

Numerous paradigms can be used to investigate learning and memory in rodents, and each has important steps that experimenters should follow. In the OFTM, it is important that maze conditions (e.g., position of maze, intra- and extra-maze cues, dim lighting and background noise level, daily time of training) are kept constant throughout the duration of the experiment. It is also necessary to minimize stress in rat subjects to ensure continuous performance in the OFTM. Stress can be kept to a minimum in female rat...

The open-field tower maze (OFTM) can be used to examine the behavioral and neurobiological mechanisms of spatial learning in rodents. Importantly, the OFTM allows researchers to study both &#8220;place-based&#8221; and &#8220;response-based&#8221; learning types. During place-learning an animal navigates through a maze using the spatial cues in the surrounding environment (e.g., a food reward is located next to a tall beacon). During response-learning the animal makes a specific turn response regardless of its position relative to spatial cues (e.g., make a left or right turn to find a food reward). It has been previously reported that response-learning and place-learning rely on two different memory systems3, 4 and mapping their neural mechanisms has been an ongoing scientific pursuit2, 5-7. The design of the OFTM was adapted from the one previously described8.
Open field paradigms (a component of the OFTM) are commonly used to investigate anxiety in rodents9. During the typical open field anxiety test rats that spend more time in the center of an open field have lower stress and anxiety levels than rats that spend more time near the perimeter. In the OFTM, a rat must enter the center of the maze to find the food reward. Therefore, correct navigation in the OFTM indicates that these rats are non-stressed. One reason why the rats are non-stressed during the training is because unlike standard open-field tests of anxiety training the OFTM requires pre-training phase during which the rat is pre-exposed to the maze over a long period of time. In addition, the training occurs in a dimly lit environment further reducing the anxiety-inducing aspect of an open field which is typically brightly lit.
Another unique feature of the OFTM, compared to some other mazes, is how learning is measured. Experimenters using the Morris Water Maze or Barnes Maze typically measure learning by analyzing the speed and/or path length a rat travels to reach its target location. These dependent measures can be affected by non-learning factors, such as changes in a rat&#8217;s general motor activity. However, the OFTM measures learning using the percent of first-choice correct response a rat makes to find the food reward. This dependent measure is not vulnerable to changes in motor performance and therefore, more accurately assesses learning than measurements of speed and distance. Nevertheless, the experimenter can also measure a rat&#8217;s speed and distance traveled in the OFTM using video tracking software.
Other maze-types, such as the T-maze, Morris Water Maze and Barnes Maze can be used to study place- and response-learning. However, the OFTM is unique in a few notable ways. First, the OFTM is a more sensitive behavioral assay than the T-maze because the OFTM has a chance performance of 25%, whereas the T-maze has a chance performance of 50%. Second, a rat can only turn in one of two ways in the T-maze, but has unrestricted route possibilities when navigating in the OFTM. Third, unlike the Morris Water Maze and Barnes Maze tasks (which have unrestricted route possibilities like the OFTM), experiments using the OFTM do not require rats to navigate through an aversive situation. Rats are motived to navigate in the Morris Water Maze to escape swimming in water and they are motivated to navigate in the Barnes Maze to avoid being in an open, brightly-lit environment. It has been demonstrated that exposure to the Morris Water Maze or Barnes Maze induces the release of stress hormone (i.e., corticosterone) in rodents10. However, rats navigate in the OFTM because they are motivated to find a food reward and training is conducted with dim ambient lighting. Thus, the OFTM is a non-aversive appetitive task. Results from experiments that use a stressful maze paradigm to examine the effects of manipulations such as drug, hormone, exercise, or diet on learning and memory mechanisms can be difficult to interpret because &#8220;stress&#8221;, by itself, may influence cognitive function11.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>OFTM construction supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Safety Glasses, Clear, Uncoated</td>
			<td>Grainger</td>
			<td>4EY97</td>
			<td>Used for general eye protection during maze construction</td>
		</tr>
		<tr>
			<td>Masking Tape, Beige, 18mm x 55m</td>
			<td>Grainger</td>
			<td>15F739</td>
			<td>General supply</td>
		</tr>
		<tr>
			<td>Wiss All-purpose Scissor</td>
			<td>Home Depot&#160;</td>
			<td>37103278647</td>
			<td>General supply</td>
		</tr>
		<tr>
			<td>Lancaster&#160; 6 foot Round Folding table - Heavy Duty white granite plastic - 29&#34;&#160;</td>
			<td>www.webstaurantstore.com</td>
			<td>384YCZ72RND</td>
			<td>Used as base of the maze</td>
		</tr>
		<tr>
			<td>Laminate Countertop sheet in White Matte Finish (48 x 96 x 0.039)</td>
			<td>Home Depot&#160;</td>
			<td>1573603504896</td>
			<td>Used as walls for the maze</td>
		</tr>
		<tr>
			<td>Laminate Countertop sheet in Black Matte Finish (48 x 96 x 0.039)</td>
			<td>Home Depot&#160;</td>
			<td>1595603504896</td>
			<td>Used as floor of the maze</td>
		</tr>
		<tr>
			<td>Adhesive, Spray, 24oz, 17.6oz Net. 3M</td>
			<td>Grainger</td>
			<td>3MA16</td>
			<td>Used to adhere the laminate floor to the table top and pieces of laminate to each other</td>
		</tr>
		<tr>
			<td>16.6 oz #90 hi-strength S</td>
			<td>Lowe&#39;s</td>
			<td>193252</td>
			<td>Used to adhere the laminate floor to the table top</td>
		</tr>
		<tr>
			<td>Superflex RTV, Silicone Adhesive Sealant, Loctile</td>
			<td>VWR</td>
			<td>300040-068</td>
			<td>Used to seal around the baze (between the floor and walls of the maze)</td>
		</tr>
		<tr>
			<td>Industrial Sealant; 10.1oz</td>
			<td>Grainger</td>
			<td>4UH06</td>
			<td>Used to seal around the baze (between the floor and walls of the maze)</td>
		</tr>
		<tr>
			<td>Painter&#39;s Masking Tape; Shurtape; 2in x 60yd</td>
			<td>Grainger</td>
			<td>6FET7</td>
			<td>Used to mark out visual patterns on maze walls</td>
		</tr>
		<tr>
			<td>Mini/Trim paint tray</td>
			<td>ACE Hardware</td>
			<td>1365105</td>
			<td>Used to hold paint&#160;</td>
		</tr>
		<tr>
			<td>Foam mini roller 4&#34; 6pk</td>
			<td>ACE Hardware</td>
			<td>1494871</td>
			<td>Used for painting patterns&#160;</td>
		</tr>
		<tr>
			<td>Val Sample Set Nbase paint</td>
			<td>ACE Hardware</td>
			<td>1505031</td>
			<td>Used as paint for pattern on the walls of the maze</td>
		</tr>
		<tr>
			<td>HP Paint Black</td>
			<td>Walmart</td>
			<td>72450405005</td>
			<td>Used to paint pattern on the walls of the maze</td>
		</tr>
		<tr>
			<td>Brush</td>
			<td>Walmart</td>
			<td>81773801110</td>
			<td>Used to paint pattern on the walls of the maze</td>
		</tr>
		<tr>
			<td>Duct Tape, 2in X 60yd, 10.5mil. Black</td>
			<td>Grainger</td>
			<td>15F767</td>
			<td>Used to wrap the exterior of the maze</td>
		</tr>
		<tr>
			<td>4X4 8ft PT pine Vinyl-CTD Post W/Cap</td>
			<td>Home Depot&#160;</td>
			<td>40933070541</td>
			<td>Used to build towers for the maze</td>
		</tr>
		<tr>
			<td>20 gram Pro Super Glue</td>
			<td>Lowe&#39;s</td>
			<td>125870</td>
			<td>Used to glue tower tops, towers to the floor of the maze and food cups to the tower tops</td>
		</tr>
		<tr>
			<td>Bead Containers 9 - 1/2</td>
			<td>Michaels</td>
			<td>652695766978</td>
			<td>Used as food cups on top of towers</td>
		</tr>
		<tr>
			<td>Spray Paint, Black, 12oz</td>
			<td>Grainger</td>
			<td>6KP22</td>
			<td>Used to paint food cups &#38; covers</td>
		</tr>
		<tr>
			<td>Room prepration</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Conair SU1W Sound Machine; 10 Sounds</td>
			<td>Home Depot&#160;</td>
			<td>204493600</td>
			<td>Used for background noise</td>
		</tr>
		<tr>
			<td>Lamp</td>
			<td>Walmart</td>
			<td>5027698392</td>
			<td>Used for dim lighting of maze room</td>
		</tr>
		<tr>
			<td>ES 14W DayLight Spiral Bulb 4pk</td>
			<td>Home Depot&#160;</td>
			<td>762148208433</td>
			<td>Used for dim lighting of maze room</td>
		</tr>
		<tr>
			<td>Shower Curtain</td>
			<td>Walmart</td>
			<td>3429910901</td>
			<td>Used for separation between the maze and the experimenter</td>
		</tr>
		<tr>
			<td>Curtain Rod</td>
			<td>Bed Bath &#38; Beyond</td>
			<td>76598645104</td>
			<td>Used for separation between the maze and the experimenter</td>
		</tr>
		<tr>
			<td>Froot Loops Cereal</td>
			<td>Walmart</td>
			<td>3800039122</td>
			<td>Used as reward</td>
		</tr>
		<tr>
			<td>Recording equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Media Recorder 3: 4 video</td>
			<td>Noldus</td>
			<td>NMR4-0200</td>
			<td>Used to record training trials</td>
		</tr>
		<tr>
			<td>Monochrome GigE Camera</td>
			<td>Noldus</td>
			<td>XCFW-BG62</td>
			<td>Used to record training trials</td>
		</tr>
		<tr>
			<td>Precision M4800 Laptop</td>
			<td>Dell</td>
			<td>N/A</td>
			<td>Used to run video software</td>
		</tr>
	</tbody>
</table>

place and response learning in the open-field tower maze

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for examining how rats learn to use a place- or response-learning to successfully navigate in an open-field arena. Additionally, this protocol describes how the OFTM differs from other behavioral maze paradigms that are commonly used to study spatial learning in rodents. The OFTM described in this article was adapted from the one previously described by Cole, Clipperton, and Walt (2007). Specifically, the OFTM was created to test spatial learning in rodents without the experimenter having to consider how &#8220;stress&#8221; might play a role as a confounding variable. Experiments have shown that stress-alone can significantly affect cognitive function1. The representative results section contains data from an experiment that used the OFTM to examine the effects of estradiol treatment on place- and response-learning in adult female Sprague Dawley rats2. Future studies will be designed to examine the role of the hippocampus and striatum in place- and response-learning in the OFTM.

The representative results described here have been previously published in the Neurobiology of Learning &#38; Memory, Elsevier2. In this experiment the OFTM was used to test the effects of chronic and cycling estradiol treatments on place- and response-learning in female Sprague-Dawley rats (250 &#8211; 300 g; N = 48). There were two important reasons why the OFTM was used to conduct this experiment. First, estradiol has been shown to induce differential effects on place- and response-learning7. Se...

Watch this Scientific Journal Video about Place and Response Learning in the Open-field Tower Maze at JoVE.com

Place and Response Learning in the Open-field Tower Maze

The Open-field Tower Maze (OFTM) is a maze created to study the behavioral and neural mechanisms of spatial learning (e.g., place- or response-learning) in rats. This maze is especially useful for experimenters who want to use a non-stressful maze paradigm to investigate spatial learning in their research.

This protocol describes how the Open-field Tower Maze (OFTM) paradigm is used to study spatial learning in rodents. This maze is especially useful for ...

place-and-response-learning-in-the-open-field-tower-maze

Research

JoVE Journal

Behavior

Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes of interest allows for enhanced temporally- and regionally-specific control of gene deletion, compared to existing methods. While conditional deletion can also be achieved by mating mice that express Cre recombinase under the control of specific gene promoters with mice carrying a floxed gene, stereotaxic microinjection allows for targeting of discrete brain areas at experimenter-determined time points of interest. In the context of cocaine conditioned place preference, and other cocaine behavioral paradigms such as self-administration or psychomotor sensitization that can involve withdrawal, extinction and/or reinstatement phases, this technique is particularly useful in exploring the unique contribution of target genes to these distinct phases of behavioral models of cocaine-induced plasticity. Specifically, this technique allows for selective ablation of target genes during discrete phases of a behavior to test their contribution to the behavior across time. Ultimately, this understanding allows for more targeted therapeutics that are best able to address the most potent risk factors that present themselves during each phase of addictive behavior.

This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.

Microinjecting recombinant adenoassociated viral (rAAV) vectors expressing Cre recombinase into distinct mouse brain regions to selectively knockout genes ...

Behavioral and Locomotor Measurements Using an Open Field Activity Monitoring System for Skeletal Muscle Diseases

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing locomotive impairment in animal models of neuromuscular disease and efficacy of therapeutic drugs that may improve locomotion and/or muscle function. The open field activity measurement provides a different measure than muscle strength, which is commonly assessed by grip strength measurements. It can also show how drugs may affect other body systems as well when used with additional outcome measures. In addition, measures such as total distance traveled mirror the 6 min walk test, a clinical trial outcome measure. However, open field activity monitoring is also associated with significant challenges: Open field activity measurements vary according to animal strain, age, sex, and circadian rhythm. In addition, room temperature, humidity, lighting, noise, and even odor can affect assessment outcomes. Overall, this manuscript provides a well-tested and standardized open field activity SOP for preclinical trials in animal models of neuromuscular diseases. We provide a discussion of important considerations, typical results, data analysis, and detail the strengths and weaknesses of open field testing. In addition, we provide recommendations for optimal study design when using open field activity in a preclinical trial.

Open field activity levels are used to assess locomotive and behavioral activity levels. This protocol provides a well-designed, standardized protocol to use in preclinical trials for neuromuscular disorders.

The open field activity monitoring system comprehensively assesses locomotor and behavioral activity levels of mice. It is a useful tool for assessing ...

Use of the Open Field Maze to Measure Locomotor and Anxiety-like Behavior in Mice

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of the most commonly used platforms to measure behaviors in animal models. It is a fast and relatively easy test that provides a variety of behavioral information ranging from general ambulatory ability to data regarding the emotionality of the subject animal. As it relates to rodent models, the procedure allows the study of different strains of mice or rats both laboratory bred and wild-captured. The technique also readily lends itself to the investigation of different pharmacological compounds for anxiolytic or anxiogenic effects. Here, a protocol for use of the open field maze to describe mouse behaviors is detailed and a simple analysis of general locomotor ability and anxiety-related emotional behaviors between two strains of C57BL/6 mice is performed. Briefly, using the described protocol we show Wild Type mice exhibited significantly less anxiety related behaviors than did age-matched Knock Out mice while both strains exhibited similar ambulatory ability.

A protocol is provided to use an Open Field Maze to access general locomotor activity, anxiety and emotionality in a laboratory mouse model.

Animal models have proven to be invaluable to researchers trying to answer questions regarding the mechanisms of behavior. The Open Field Maze is one of ...

The Double-H Maze: A Robust Behavioral Test for Learning and Memory in Rodents

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their behavioral flexibility and required memory duration, the number of goals and pathways, and also the overall task complexity. A confounding feature in many of these tasks is the lack of control over the strategy employed by the rodents to reach the goal, e.g., allocentric (declarative-like) or egocentric (procedural) based strategies. The double-H maze is a novel water-escape memory task that addresses this issue, by allowing the experimenter to direct the type of strategy learned during the training period. The double-H maze is a transparent device, which consists of a central alleyway with three arms protruding on both sides, along with an escape platform submerged at the extremity of one of these arms.
Rats can be trained using an allocentric strategy by alternating the start position in the maze in an unpredictable manner (see protocol 1; &#167;4.7), thus requiring them to learn the location of the platform based on the available allothetic cues. Alternatively, an egocentric learning strategy (protocol 2; &#167;4.8) can be employed by releasing the rats from the same position during each trial, until they learn the procedural pattern required to reach the goal. This task has been proven to allow for the formation of stable memory traces.
Memory can be probed following the training period in a misleading probe trial, in which the starting position for the rats alternates. Following an egocentric learning paradigm, rats typically resort to an allocentric-based strategy, but only when their initial view on the extra-maze cues differs markedly from their original position. This task is ideally suited to explore the effects of drugs/perturbations on allocentric/egocentric memory performance, as well as the interactions between these two memory systems.

The goal of this protocol is to investigate spatial cognition in rodents. The double-H water maze is a novel test, which is particularly useful to elucidate the different components of learning, consolidation and memory, as well as the interplay of memory systems.

Spatial cognition research in rodents typically employs the use of maze tasks, whose attributes vary from one maze to the next. These tasks vary by their ...

Quantifying Learning in Young Infants: Tracking Leg Actions During a Discovery-learning Task

Task-specific actions emerge from spontaneous movement during infancy. It has been proposed that task-specific actions emerge through a discovery-learning process. Here a method is described in which 3-4 month old infants learn a task by discovery and their leg movements are captured to quantify the learning process. This discovery-learning task uses an infant activated mobile that rotates and plays music based on specified leg action of infants. Supine infants activate the mobile by moving their feet vertically across a virtual threshold. This paradigm is unique in that as infants independently discover that their leg actions activate the mobile, the infants&#8217; leg movements are tracked using a motion capture system allowing for the quantification of the learning process. Specifically, learning is quantified in terms of the duration of mobile activation, the position variance of the end effectors (feet) that activate the mobile, changes in hip-knee coordination patterns, and changes in hip and knee muscle torque. This information describes infant exploration and exploitation at the interplay of person and environmental constraints that support task-specific action. Subsequent research using this method can investigate how specific impairments of different populations of infants at risk for movement disorders influence the discovery-learning process for task-specific action.

A method is described in which 3-4 month old infants learn a task by discovery and their leg movements are captured to quantify the learning process.

Task-specific actions emerge from spontaneous movement during infancy. It has been proposed that task-specific actions emerge through a discovery-learning ...

Reinstatement of Drug-seeking in Mice Using the Conditioned Place Preference Paradigm

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to acquire a conditioned place preference in a drug-paired compartment, and after the post-conditioning test, they perform several sessions to extinguish the established preference. The CPP permits the evaluation of the conditioned rewarding effects of drugs related to environmental cues. Then, the extinguished CPP can be robustly reinstated by the non-contingent administration of a priming dose of the drug, and by exposure to stressful stimuli. Both methods will be explained here. When the animal reinitiates the behavioral response, a reinstatement of the conditioned reward is considered to have taken place.
The main advantages of this protocol are that it is non-invasive, inexpensive, and simple with good validity criteria. In addition, it allows the study of different environmental manipulations, such as stress or diet, which can modulate relapse into drug seeking behaviors. However, one limitation is that if the researcher aims to explore the motivation and primary reinforcing effects of the drug, it should be complemented with self-administration procedures, as they involve operant responses of animals.

This protocol describes the Conditioned Place Preference (CPP) as a model of relapse. This procedure permits the measurement of relapse in laboratory animals, considering the impact of drug-associated environmental cues as craving and relapse in abstaining addicts is currently the focus of drug-abuse treatment programs.

The present protocol describes the Conditioned Place Preference (CPP) as a model of relapse in drug addiction. In this model, animals are first trained to ...

Analyzing Spatial Learning and Prosocial Behavior in Mice Using the Barnes Maze and Damsel-in-Distress Paradigms

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. The Barnes maze can also assess other mouse behaviors, such as general motivation to escape from the maze platform and exploratory behavior. The Barnes maze can measure whether a genetic mutation or environmental variable can impact the acquisition and retention of spatial memories, as well as provide information about the search strategy employed by the mice. Here we use the Barnes maze to detect a memory deficit in adult mice following a single developmental ethanol exposure event. The newly described Damsel-in-Distress paradigm exposes a male mouse to a female mouse trapped in a chamber in the open center field of the arena. It provides an opportunity for the mouse to socially respond to the trapped female and exhibit prosocial behavior. The Damsel-in-Distress paradigm can also be used to examine mouse behavior in a novel arena and measure locomotor activity. Both the Barnes Maze and the Damsel-in-Distress protocols require minimal financial investment and most aspects of the tests can be constructed from common lab supplies. These flexible and accessible tools can also be used to detect behavioral changes over the course of development.

This protocol measures spatial learning and memory using the Barnes maze. A novel Damsel-in-Distress paradigm is used to assess locomotor activity and prosocial behavior in mice.

The Barnes maze is a reliable measure of spatial learning and memory that does not require food restriction or exposure to extremely stressful stimuli. ...

Analysis of Learning and Memory Ability in an Alzheimer's Disease Mouse Model using the Morris Water Maze

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific research to assess the learning and memory of animals. Due to the extensive use of the MWM test, visual experimental protocols are essential for researchers. This manuscript uses the latest studies to introduce the protocol of the MWM test. Alzheimer&#8217; Disease (AD) is characterized by a progressive loss of memory and cognitive function. An alternative and complementary treatment used for AD is Manual Acupuncture (MA). To assess the learning and memory ability of AD model mice, the MWM test was conducted. The visible platform trial, hidden platform trial, probe trial, and reversal trial of MWM were used to evaluate spatial learning and memory ability. In the visible platform trial, the swimming speed and escape latency of mice in different groups was not significantly different. In the hidden platform and reversal trials, the AD group showed a long escape latency. The escape latency decreased significantly after the MA treatment. Low platform crossover number and the proportion of time in the SW quadrant in the probe trial increased after the MA treatment (p &#60; 0.05 or p &#60; 0.01). The results of the MWM tests suggest that MA can effectively improve the spatial learning and memory abilities of AD model mice. Rigorous experimental operations provided assurance of the reliability of the results.

Herein, a protocol to conduct the Morris water maze tests to evaluate the ability of learning and memory of Alzheimer&#8217;s Disease model mice and to assess the effect of manual acupuncture for treating them is described.

A Morris water maze (MWM) experiment forces experimental animals to swim and learn to find a platform hidden in the water. It is widely used in scientific ...

Chronic Stress Shifts Effort-Related Choice Behavior in a Y-Maze Barrier Task in Mice

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that measures motivation to expend effort and obtain reward. In mice, chronic stress exposure significantly impacts motivation to work for a higher value reward when a lesser value reward is freely available compared to unstressed mice. Here we describe the chronic corticosterone administration paradigm, which produces a shift in effortful responding in the Y-maze barrier task. In the Y-maze task, one arm contains 4 food pellets, while the other arm contains only 2 pellets. After mice learn to select the high reward arm, barriers with progressively increasing height are then introduced into the high reward arm over multiple test sessions. Unfortunately, most chronic stress paradigms (including corticosterone and social defeat) were developed in male mice and are less effective in female mice. Therefore, we also discuss chronic non-discriminatory social defeat stress (CNSDS), a stress paradigm we developed that is effective in both male and female mice. Repeating results with multiple distinct chronic stressors in male and female mice combined with increased usage of translationally relevant behavior tasks will help to advance the understanding of how chronic stress can precipitate mood disorders.

The Y-maze barrier task is a behavior test that examines motivation to expend effort for reward. Here, we discuss testing multiple well-validated chronic stressors including chronic corticosterone and social defeat stress with this behavior, as well as the novel chronic non-discriminatory social defeat stress (CNSDS), which is effective in females.

Mood disorders, including major depressive disorder, can be precipitated by chronic stress. The Y-maze barrier task is an effort-related choice test that ...

Behavioral Assessment of Visual Function via Optomotor Response and Cognitive Function via Y-Maze in Diabetic Rats

The optomotor response and the Y-maze are behavioral tests useful for assessing visual and cognitive function, respectively. The optomotor response is a valuable tool to track changes in spatial frequency (SF) and contrast sensitivity (CS) thresholds over time in a number of retinal disease models, including diabetic retinopathy. Similarly, the Y-maze can be used to monitor spatial cognition (as measured by spontaneous alternation) and exploratory behavior (as measured by a number of entries) in a number of disease models that affect the central nervous system. Advantages of the optomotor response and the Y-maze include sensitivity, speed of testing, the use of innate responses (training is not needed), and the ability to be performed on awake (non-anesthetized) animals. Here, protocols are described for both the optomotor response and the Y-maze and examples of their use shown in models of Type I and Type II diabetes. Methods include preparation of rodents and equipment, performance of the optomotor response and the Y-maze, and post-test data analysis.

Neural degeneration in both eyes and brain as a result of diabetes can be observed through behavioral tests carried out on rodents. The Y-maze, a measure of spatial cognition, and the optomotor response, a measure of visual function, both provide insight into potential diagnoses and treatments.

The optomotor response and the Y-maze are behavioral tests useful for assessing visual and cognitive function, respectively. The optomotor response is a ...