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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Working Memory Water Maze
<ol>
	<li>Equipment
	<ol>
		<li>Use a tank filled with water to a height of 28 cm and maintained at 26 &#177; 1 &#176;C.</li>
		<li>Use a clear acrylic glass platform that is 10 cm in diameter on a stand 26 cm in height. 
		NOTE: The top surface of the platform should be coated with silicon in the shape of a circle with an X across it. This allows the rats to climb up onto the platform and gives them traction so they do not slip off.</li>
		<li>Gather a stopwatch, a heat lamp, disposable towels, absorbent pads, extra cages, and a small, long-handled aquarium fish net. Use a computerized video tracking system that is connected to a video camera to record the rat swimming and send the data to the computer. Save the video and data on the computer for later analysis.</li>
	</ol>
	</li>
	<li>Working Memory Water Maze Testing
	<ol>
		<li>Give the rats four pairs of trials each day for five consecutive days, place the platform in each of the four quadrants and start the rats from each of the four starting points (N, S, E, W) as described below.</li>
		<li>First, define the starting location-platform pairs to be used throughout the experiment. 
		NOTE: The order of the quadrants where the platform is located and the starting point used needs to be in a different order for each of the five days of swimming, but the same for each rat.
		<ol>
			<li>Use four starting points (N, S, E, or W) and four platform locations (Quadrants 1, 2, 3, or 4;&#160;Figure 1). For example, (N, 2; E, 4; S, 1; W, 3; see&#160;Figure 1). Plan a balanced order (not random) to avoid starting points too close to the platforms (No starting point is the same quadrant as the platform location). Set up a data sheet using the platform quadrants and four starting points.</li>
			<li>Write a protocol for the video tracking software to use in order to video the rats swimming and to collect specified data (e.g., duration of swim, speed, distance traveled before finding the platform). 
			NOTE: The tracking software will automatically stop recording after the specified duration. The protocol should allow for specifying where the platform is placed, how many trials to run per animal, and how many animals will be tested per session, and also the maximum duration allowed (e.g., 120 s).</li>
			<li>Test 4&#8211;6 rats per session. 
			NOTE: More than 6 rats create an issue in the timing between rats and can lead to error by the handler. The warming boxes also become crowded.</li>
		</ol>
		</li>
		<li>Trial 1
		<ol>
			<li>Open the video tracking software and load the correct protocol including the water maze map.</li>
			<li>Place the platform in the assigned location (e.g., 2;&#160;Figure 1) and check that it matches the map in the software. Prepare the tracking software to start when the rat enters the field of view of the video camera.</li>
			<li>Place the rat in the tank facing the wall at the assigned location (e.g., N;&#160;Figure 1) and immediately start the timer.</li>
			<li>Allow the rat 120 s to find the platform. When the rat finds the platform, stop the timer and record the time on the worksheet. If the rat fails to find the platform, lead it to the platform by hand and record 120 s. Allow the rat 15 s to remain on the platform.</li>
		</ol>
		</li>
		<li>Trial 2
		<ol>
			<li>Check that the software is ready for Trial 2. Place the rat back in the tank at the same starting position (N). Repeat step 1.2.3.4.</li>
		</ol>
		</li>
		<li>After Trial 2, place the rat in the heated enclosure for 4 min. Move the platform to the second location (4;&#160;Figure 1), and check that it matches the map in the software.</li>
		<li>Repeat Trial 1 and 2 procedures (steps 1.2.3&#8211;1.2.4) until all four starting location/platform pairings are completed.</li>
	</ol>
	</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr >
			<td >Sprague-Dawley rats</td>
			<td >Charles Rivers Laboratories</td>
			<td >CD-IGS rats&#160; strain code 001</td>
			<td >Male albino 300-350g at arrival</td>
		</tr>
		<tr >
			<td >Fiberglass pool</td>
			<td>Manufacturer unknown</td>
			<td></td>
			<td>72&#34; diameter x 30&#34; deep (~ 500 gal)</td>
		</tr>
		<tr >
			<td >Plexiglass platform</td>
			<td>Hand-made by Maggie Parsley</td>
			<td></td>
			<td>10 cm diameter&#160; 26&#34; tall with silicone applied to the surface of the platform to provide a gripping surface</td>
		</tr>
		<tr >
			<td >Plexiglass animal boxes w/ lids</td>
			<td>UTMB Machine Shop</td>
			<td></td>
			<td>2 boxes&#160; 10&#34; w x 16&#34; L x 9&#34; h</td>
		</tr>
		<tr >
			<td >Spot lights/heat lamps</td>
			<td>Home Depot</td>
			<td></td>
			<td>3 around pool&#160; 2 over boxes to dry animals</td>
		</tr>
		<tr >
			<td >AnyMaze</td>
			<td>San Diego Instruments</td>
			<td>9001</td>
			<td><a href="http://www.sandiegoinstruments.com/any-maze-video-tracking/">http://www.sandiegoinstruments.com/any-maze-video-tracking/</a></td>
		</tr>
	</tbody>
</table>

the water maze with platform relocation: a method to assess working memory

Source: Hausser, N., et al. <a href="https://www.jove.com/video/56044/detecting-behavioral-deficits-in-rats-after-traumatic-brain-injury">Detecting Behavioral Deficits in Rats After Traumatic Brain Injury.</a>&#160;J. Vis. Exp (2018).
In this video, we describe the Morris water maze test with platform relocation, a navigational task used to study working memory in rodents.

<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/20049/20049_Figure1.jpg" /> 
Figure 1. Diagram of the water maze.&#160;This diagram demonstrates the possible platform locations (1, 2, 3, 4) and starting points (N, S, E, W) for the working memory Morris water maze. Rats are allowed two trials from each starting location/platform pairing. There is a 15 s inter-trial interval and 4 min rest in a warming chamber between pairs of trials for a total of four pair...

The Water Maze with Platform Relocation: A Method to Assess Working Memory

Source: Hausser, N., et al. Detecting Behavioral Deficits in Rats After Traumatic Brain Injury.&#160;J. Vis. Exp (2018).
In this video, we describe the ...

Ensure visual cues are placed around the water maze tank to help subjects learn the location of the submerged hidden platform. Place the subject in the tank away from the platform facing the wall. The animal will swim to locate the stage and escape.
End the trial after the animal finds the platform or a predetermined time has elapsed. Let the subject stay on top of the platform before repeating the trial a second time. If it fails to locate the stage, guide the animal by hand and allow it to remain on the platform for the duration of the trial.
Then, to assess the animals working memory, repeat the trial a second time. Subjects will have to remember how to locate the platform and use the visual cues as reference. Record the time and path length for repeated trials to use as measures of working memory. After the animal has rested, continue by moving the platform to a new location and perform trials 1 and 2.

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JoVE Encyclopedia of Experiments

Biology

Rodent Models

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1.4K Views. Source: Hausser, N., et al. Detecting Behavioral Deficits in Rats After Traumatic Brain Injury. J. Vis. Exp (2018). In this video, we describe the Morris water maze test with platform relocation, a navigational task used to study working memory in rodents. 

Video: The Water Maze with Platform Relocation: A Method to Assess Working Memory - Concept

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to the mouse becoming increasingly important. But researchers found that some strains of mutant mice were prone to problems like passively floating or diving when they were tested in the Morris water maze11. This was unsurprising considering their natural habitat; rats swim naturally (classically, the "sewer rat"), whereas mice evolved in the dry areas of central Asia.
To overcome these problems, it was considered whether shallow water would be a sufficient stimulus to provide escape motivation for mice. This would also avoid the problems of drying the small creatures with a towel and then putting them in a heated recovery chamber to avoid hypothermia, which is a much more serious problem than with rats; the large ratio of surface area to volume of a mouse makes it particularly vulnerable to rapid heat loss.
Another consideration was whether a more natural escape strategy could be used, to facilitate learning. Since animals that fall into water and swim away from the safety of the shore are unlikely to pass on their genes, animals have evolved a natural tendency to swim to the edge of a body of water. The Morris water maze, however, requires them to swim to a hidden platform towards the center of the maze - exactly opposite to their evolved behavior. Therefore the paddling maze should incorporate escape to the edge of the apparatus. This feature, coupled with the use of relatively non-aversive shallow water, embodies the "Refinement" aspect of the "3 Rs" of Russell and Burch8.
Various types of maze design were tried; the common feature was that the water was always shallow (2 cm deep) and escape was via a tube piercing the transparent wall of the apparatus. Other tubes ("false exits") were also placed around the walls but these were blocked off. From the inside of the maze all false exits and the single true exit looked the same. Currently a dodecagonal (12-sided) maze is in use in Oxford, with 12 true/false exits set in the corners. In a recent development a transparent paddling Y-maze has been tested successfully.

Shallow Water Paddling Variants of Water Maze Tests in Mice

Mice can swim, but many strains appear to find this activity stressful. To overcome this problem mazes have been devised where escape from shallow water is used to motivate behaviour. These have been demonstrated to support learning at least as good as the traditional and widely used Morris water maze.

When Richard Morris devised his water maze in 19817, most behavioral work was done in rats. However, the greater understanding of mouse genetics led to ...

JoVE Journal

Behavior

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 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 ...

Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor locomotion after surgery. For example, rats who undergo surgical procedures and are subjected to the Morris water maze may not be able to swim, thus voiding the experiment.
New Method: We established a modified behavioral spontaneous alternation T-maze test. The major advantage of the modified T-maze protocol is its relatively simple design that is powerful enough to assess functional learning/memory after ischemia. Additionally, the data analysis is simple and straightforward. We used the T-maze to determine the rats&#39; learning/memory deficits both in the presence or absence of mild to moderate (6 min) asphyxial cardiac arrest (ACA). Rats have a natural tendency for exploration and will explore the alternate arms in the T-maze, whereas hippocampal-lesioned rats tend to adopt a side-preference resulting in decreased spontaneous alternation ratios, revealing the hippocampal-related functional learning/memory in the presence or absence of ACA.
Results: ACA groups have higher side-preference ratios and lower alternations as compared to control.
Comparison with Existing Method(s): The Morris water and Barnes maze are more prominent for assessing learning/memory function. However, the Morris water maze is more stressful than other mazes. The Barnes maze is widely used to measure reference (long-term) memory, while ACA-induced neurocognitive deficits are more closely related to working (short-term) memory.
Conclusions: We have developed a simple, yet effective strategy to delineate working (short-term) memory via the T-maze in our global cerebral ischemia model (ACA).

This protocol describes the use of a modified T-maze to evaluate functional learning/memory in asphyxia cardiac arrest-induced cerebral ischemia.

Background: Evaluating mild to moderate cognitive impairment in a global cerebral ischemia (i.e. cardiac arrest) model can be difficult due to poor ...

The Water Maze with Platform Relocation: A Method to Assess Working Memory

Transcript

The Water Maze with Platform Relocation: A Method to Assess Working Memory

Shallow Water (Paddling) Variants of Water Maze Tests in Mice

Place and Response Learning in the Open-field Tower Maze

Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest