Name: morris water maze test for learning and memory deficits in alzheimer's disease model mice
Uploaded: 2011-07-20T13:00:00.000+00:00
Duration: 7 min 2 s
Description: Watch this Scientific Journal Video about Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice at JoVE.com

This work was supported by the Canadian Institutes of Health Research (CIHR), the Townsend Family, and Jack Brown and Family Alzheimer&#x2019;s Research Foundation (to W. S.). W. S. is the holder of the Canada Research Chair in Alzheimer's Disease. P.L. was supported by a NSERC Alexander Graham Bell Canada Graduate Scholarship Doctoral Research Award and a Michael Smith Foundation for Health Research Senior Graduate Studentship.

1. Preparation
<ol>
<li>Equipment preparation<ol>
 <li>Obtain a circular pool with a diameter of 150cm and a depth of 50cm (Fig. 1). If using black mice, a white pool should be used; if using white mice, a black pool should be used.</li>
 <li>Arrange the room such that the animal being tested cannot see the experimenter during testing. This can be accomplished with drapes or room dividers.</li>
 <li>Place high contrast spatial cues about the room, and/or on the interior of the pool at a location which would be above the water surface.</li>
 <li>Place a 10cm diameter platform in the pool – white for a white pool, clear plexiglass for a black pool. Fill the pool with water until the platform is 1cm above the water surface. Let the water equilibrate to room temperature (22 °C). Depending on the water temperature this may take one to three days, or alternatively hot water can be added to speed up the equilibration.</li></ol></li>

 <li>Software preparation<ol>
 <li>Calibrate the pool in the computer software so the camera can create physical distance information from pixel-based information. Divide the pool into 4 quadrants. Specify the platform zone as a variable zone which can change with each trial. Create 5 platform subzones – one in each quadrant, and one in the center of the pool. Save the calibration and use it for the remaining test days. (See example Fig. 2).</li>
 <li>Set the maximum trial time as 60 sec. If the mouse finds the platform before this time, program the software to stop the trial when the platform is found.</li>
 <li>Specify the program to begin tracking automatically, when the experimenter exits the testing area. Utilize any “reflection minimization” options your software package provides.</li>
 <li>Track path length, escape latency, and time spent in each quadrant.</li></ol></li>

</ol>
2. Day 1: Visible Platform
<ol>
 <li>Computer Program<ol>
 <li>Load the pool calibration into the tracking software.</li>
 <li>Create 5 trials, with an inter-trial interval appropriate for your experiment. Program the platform location and starting direction to differ with each trial. See Table 1 for an example protocol.</li></ol></li>
 <li>Testing procedure<ol>
 <li>Transfer the mice from their housing facility to the behavior room. Keep the mice in an area where they cannot see the pool or spatial cues. Let them adjust to the new environment for at least 30 minutes before testing.</li>
 <li>Place a flag on the platform to increase its visibility.</li>
 <li>To begin testing, lift mouse from the home cage by the base of the tail. Support the mouse as you bring it to the testing area. Lifting the mouse by the base of the tail, gently place the mouse into the water, facing the edge of the pool. Quickly leave the testing area.</li>
 <li>If the mouse finds the platform before the 60 sec cut-off, allow the mouse to stay on the platform for 5 seconds then return it to its home cage. If the mouse does not find the platform, place the mouse on the platform and allow it to stay there for 20 sec before returning it to its home cage.</li> 
 <li>Repeat for all mice in the trail. Begin each subsequent trial with a different platform location and starting direction, as you have programmed into your software.</li>
 <li>When testing is complete, return the mice to their housing facility. Mice are dried off and normothermia is assured prior to returning to animal facility.</li>
 <li>In preparation for the following day, remove the flag from the platform and add additional water to the pool to submerge the platform to 1cm below the surface. </li></ol></li>

</ol>
3. Days 2-5: Hidden Platform
<ol>
 <li>Computer Program<ol>
 <li>Load the pool calibration into the tracking software.</li>
 <li>Create 5 trials, with an inter-trial interval appropriate for your experiment. Program the platform location to remain in the same position throughout all trials and days, but have the starting direction differ with each trial, each day.</li></ol></li>
 <li>Testing procedure<ol>
 <li>For black mice, add non-toxic, white, powdered tempera paint to the pool and mix thoroughly. Use enough paint such that the submerged platform is not visible from the surface of the water. For white mice, a black pool with clear water and a clear plexiglass platform should be used.</li>
 <li>Follow steps 2.2.3 to 2.2.6.</li></ol></li>
</ol>
4. Day 6: Probe Trial
<ol>
 <li>Computer Program<ol>
 <li>Load the pool calibration into the tracking software.</li>
 <li>Create 1 trial with no platform zone, and one starting direction. The starting direction farthest from the platform quadrant used on days 2-5 is preferred. Set the trail length to 60 sec.</li></ol></li>
 <li>Testing procedure<ol>
 <li>Remove the platform from the pool.</li>
 <li>Follow steps 2.2.3 to 2.2.6.</li>
 </ol></li>
</ol>
5. Data analysis
<ol>
<li>For each day and each mouse, average the 5 trials to give a single path length and escape latency for each test subject. Calculate the combined error appropriately. For day 6, simply collect the path length, escape latency, and time spent in the platform quadrant for each mouse.</li>
<li>If any differences exist between groups on Day 1, it is likely a problem with vision rather than learning and memory. Only proceed with analysis if no differences are seen on day 1.</li>
<li>Compare the learning curves for Days 2-5 using statistics appropriate for your data set. A steeper curve represents faster task acquisition; a shallower curve represents a deficit in task acquisition. The data from day 2 to day 5 are analyzed using ANOWA.</li>
<li>For day 6, compare the percent of time spent in the previously learned platform quadrant, using statistics appropriate for your data set. A higher percentage of time spent in the platform quadrant is interpreted as a higher level of memory retention.</li>
</ol>
6. Representative Results
We have used the Morris Water Maze test to examine hypoxia’s effect on AD pathogenesis (7) and valproic acid (VPA)’s pharmaceutical potential for AD treatment (6) in transgenic AD model mice. Figure 3 is the representative result we reported in our study on VPA’s effect on memory deficits in the APP23 AD mouse model (6). On day 1 (visible platform trials), there is no difference between the VPA treated and control groups in latency (Fig. 3A) and path length (Fig. 3A) indicating that both of the groups have similar motor and visual capabilities. From this we assume the mice are able to see the flagged-platform and the cues in surrounding environment, and can swim acceptably. For days 2-5 (Day 1 to 4 of hidden platform trials) the example shows a difference in the escape latency (Fig. 3C) and path length (Fig 3D) between the groups, suggesting that VPA treated mice performed significantly better than controls over time. The probe trail results on the last day (Day 6) show that the number of times the mice traveled into the third quadrant, where the hidden platform was previously placed, was significantly greater with VPA treatment compared to control (Fig. 3E). These data indicate that VPA treatment significantly improves the memory deficits seen in APP23 mice.
<img src="/files/ftp_upload/2920/2920fig1.jpg" alt="Static equilibrium; water tank experiment; fluid dynamics; stability analysis; educational research." data-altupdatedat="1747911167000" data-alt="Figure 1"> 
Figure 1. Equipment setup for the Morris Water Maze visible platform test day. The pool is shielded from the experimenter using room dividers. Spatial cues are located on the walls, and maybe placed on the interior of the pool, above the water surface, if desired. The pool is filled with clear water, with the platform located 1cm above the surface. A flag has been placed on the platform to enhance visibility. 
<img src="/files/ftp_upload/2920/2920fig2.jpg" alt="Petri dish analysis in software interface; microbial growth imaging; data processing setup." data-altupdatedat="1747911167000" data-alt="Figure 2"> 
Figure 2. Screen capture from the Any-Maze™ Video Tracking System demonstrating pool calibration. The pool is viewed from above by a black and white analog tracking camera with a RTV24 Digitizer. Several zones are defined within the software and the total pool is divided into 4 quadrants. A fifth, platform zone is entered which can vary across trials, with five possible locations: NW, NE, SW, SE, or Center. A calibration line (ticked line across center) is added to allow the software to convert pixel distances into physical distances. 
<img src="/files/ftp_upload/2920/2920fig3.jpg" alt="Comparative behavior analysis graphs; escape latency, path length, passing times; control vs VPA." data-altupdatedat="1747911167000" data-alt="Figure 3"> 
Figure 3. Representative results for the Morris Water Maze. The 7-month APP23 transgenic mice carrying human Swedish mutant APP gene were tested after one month of daily VPA (n=30 mice) or vehicle solution (n=30 mice) injections. (A) During the first day of visible platform tests, the VPA treated and control APP23 mice exhibited a similar latency to escape onto the visible platform. P&gt;0.05 by student’s t-test. (B) The VPA-treated and control APP23 mice had similar swimming distances before escaping onto the visible platform in the visible platform test. P&gt;0.05 by student’s t-test. (C) In hidden platform tests, VPA treated APP23 mice showed a shorter latency to escape onto the hidden platform on the 3rd and 4th day, P&lt;0.001 by ANOVA. (D) The VPA-treated APP23 mice had a shorter swimming length before escaping onto the hidden platform on the 3rd and 4th day, P&lt; 0.01 by ANOVA. (E) In the probe trial on the 6th day, the VPA-treated APP23 mice traveled into the third quadrant, where the hidden platform was previously placed, significantly more times than controls. * P&lt;0.005 by student’s t-test. (Adapted and reprinted from The Journal of Experimental Medicine 205, 2781-2789, 2008, Rockefeller University Press, Originally published in J. Exp. Med. doi:10.1084/jem.20081588.) (6).



<table border="1">
 <tbody>
 <tr>
 <td> </td>
 <td colspan="2">Day 1</td> 
 <td>Day 2</td>
 <td>Day 3</td>
 <td>Day 4</td>
 <td>Day 5</td>
 <td>Day 6</td>
 </tr>
 <tr>
 <td> </td>
 <td>Platform location</td>
 <td>Starting Direction</td>
 <td colspan="4">Platform Location: SW
Starting Location as follows:
</td>
 <td>No platform.</td>
 </tr>
 <tr>
 <td>Trial 1</td>
 <td>SW</td>
 <td>S</td>
 <td>W</td>
 <td>N</td>
 <td>N</td>
 <td>E</td>
 <td>N</td>
 </tr>
 <tr>
 <td>Trial 2</td>
 <td>NW</td>
 <td>N</td>
 <td>S</td>
 <td>W</td>
 <td>E</td>
 <td>S</td>
 <td> </td>
 </tr>
 <tr>
 <td>Trial 3</td>
 <td>NE</td>
 <td>S</td>
 <td>N</td>
 <td>E</td>
 <td>W</td>
 <td>W</td>
 <td> </td> 
 </tr>
 <tr>
 <td>Trial 4</td>
 <td>Center</td>
 <td>E</td>
 <td>E</td>
 <td>W</td>
 <td>S</td>
 <td>E</td>
 <td> </td>
 </tr>
 <tr>
 <td>Trial 5</td>
 <td>SE</td>
 <td>W</td>
 <td>S</td>
 <td>S</td>
 <td>N</td>
 <td>N</td> 
 <td> </td> 
 </tr>
 </tbody>
</table>
Table 1. Sample water maze protocol* 
* Note how both the platform position and starting direction change on day 1, whereas on days 2-5 the platform position remains constant while the starting direction changes. On day 6, there is no platform and a single trial. The starting direction for day 6 is farthest from the previous platform location (SW) so that the mice must travel some distance before entering the previously learned platform quadrant.

<ol>
	<li>Morris, R. G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+localization+does+not+require+the+presence+of+local+cues.">Spatial localization does not require the presence of local cues.</a> Learning and Motivation. 12, 239-260 (1981).</li><li>O'Keefe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+the+hippocampal+place+cells.">A review of the hippocampal place cells.</a> Prog Neurobiol. 13, 419-439 (1979).</li><li>Scoville, W. B., Milner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+recent+memory+after+bilateral+hippocampal+lesions.">Loss of recent memory after bilateral hippocampal lesions.</a> J Neurol Neurosurg Psychiatry. 20, 11-21 (1957).</li><li>Eichenbaum, H., Stewart, C., Morris, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hippocampal+representation+in+place+learning.">Hippocampal representation in place learning.</a> J Neurosci. 10, 3531-3542 (1990).</li><li>Block, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+ischemia+and+behavioural+deficits.">Global ischemia and behavioural deficits.</a> Progress in Neurobiology. 58, 279-295 (1999).</li><li>Qing, H., He, G., Ly, P. T., Fox, C. J., Staufenbiel, M., Cai, F., Zhang, Z., Wei, S., Sun, X., Chen, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Valproic+acid+inhibits+Abeta+production,+neuritic+plaque+formation,+and+behavioral+deficits+in+Alzheimer's+disease+mouse+models.">Valproic acid inhibits Abeta production, neuritic plaque formation, and behavioral deficits in Alzheimer's disease mouse models.</a> J Exp Med. 205, 2781-2789 (2008).</li><li>Sun, X., He, G., Qing, H., Zhou, W., Dobie, F., Cai, F., Staufenbiel, M., Huang, L. E., Song, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia+facilitates+Alzheimer's+disease+pathogenesis+by+up-regulating+BACE1+gene+expression.">Hypoxia facilitates Alzheimer's disease pathogenesis by up-regulating BACE1 gene expression.</a> Proc Natl Acad Sci U S A. , 18727-18732 (2006).</li><li>D'Hooge, R., De Deyn, P. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+the+Morris+water+maze+in+the+study+of+learning+and+memory.">Applications of the Morris water maze in the study of learning and memory.</a> Brain Research Reviews. 36, 60-90 (2001).</li><li>Brandeis, R., Brandys, Y., Yehuda, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+the+Morris+Water+Maze+in+the+study+of+memory+and+learning.">The use of the Morris Water Maze in the study of memory and learning.</a> Int J Neurosci. 48, 29-69 (1989).</li><li>Lipp, H. P., Wolfer, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetically+modified+mice+and+cognition.">Genetically modified mice and cognition.</a> Curr Opin Neurobiol. 8, 272-280 (1998).</li><li>Sandi, 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+role+and+mechanisms+of+action+of+glucocorticoid+involvement+in+memory+storage.">The role and mechanisms of action of glucocorticoid involvement in memory storage.</a> Neural Plast. 6, 41-52 (1998).</li></ol>

All experiments were conducted in accordance with The University of British Columbia Animal Care and Use committee and CIHR guidelines.

Age, sex, species, and strain differences influence MWM performance (8). Studies indicate that aged mice have poor performance in the MWM, while male rodents perform better than females; additionally, floating is more pronounced in mice than rats (9, 10). Therefore, these elements should be equated across all tests. Evidence also suggests that stressed animals perform more poorly in the MWM (11), thus environmental factors which may cause stress, such as temperature, light, and noise, should be monitored and kept constant over the task.

<table><tbody><tr><td>AnyMaze Video Tracking System</td><td>Stoelting Co.</td><td></td><td></td></tr><tr><td>Tempera Paint</td><td>Reeves Poole Groups</td><td></td><td>White, powdered</td></tr></tbody></table>

morris water maze test for learning and memory deficits in alzheimer&#39;s disease model mice

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including acquisition of spatial memoryand long-term spatial memory 1. The MWM is a relatively simple procedure typically consisting of six day trials, the main advantage being the differentiation between the spatial (hidden-platform) and non-spatial (visible platform) conditions 2-4. In addition, the MWM testing environment reduces odor trail interference 5. This has led the task to be used extensively in the study of the neurobiology and neuropharmacology of spatial learning and memory. The MWM plays an important role in the validation of rodent models for neurocognitive disorders such as Alzheimer&#x2019;s Disease 6, 7. In this protocol we discussed the typical procedure of MWM for testing learning and memory and data analysis commonly used in Alzheimer&#x2019;s disease transgenic model mice.

Watch this Scientific Journal Video about Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice at JoVE.com

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice

The Morris Water Maze is a behavioral task to test hippocampal-dependent learning and memory. It has been widely used in the study of neurobiology, neuropharmacology and neurocognitive disorders in rodent models.

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#39;s Disease Model Mice

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including ...

morris-water-maze-test-for-learning-memory-deficits-alzheimer-s

Research

JoVE Journal

Neuroscience

127.1K Views.  University of British Columbia. The Morris Water Maze is a behavioral task to test hippocampal-dependent learning and memory. It has been widely used in the study of neurobiology, neuropharmacology and neurocognitive disorders in rodent models.

Video: Morris Water Maze Test for Learning and Memory Deficits in Alzheimer's Disease Model Mice

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 ...

Behavior

Low-stress Route Learning Using the Lashley III Maze in Mice

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other aversive stimuli to motivate task acquisition. As animals age, sensory modalities deteriorate. For example, many strains of mice develop hearing deficits or cataracts. Changes in the sensory systems required to guide mice during task acquisition present potential confounds in interpreting learning changes in aging animals. Moreover, the use of aversive stimuli to motivate animals to learn tasks is potentially confounding when comparing mice with differential sensitivities to stress. To minimize these types of confounding effects, we have implemented a modified version of the Lashley III maze. This maze relies on route learning, whereby mice learn to navigate a maze via repeated exposure under low stress conditions, e.g. dark phase, no food/water deprivation, until they navigate a path from the start location to a pseudo-home cage with 0 or 1 error(s) on two consecutive trials. We classify this as a low-stress behavior test because it does not rely on aversive stimuli to encourage exploration of the maze and learning of the task. The apparatus consists of a modular start box, a 4-arm maze body, and a goal box. At the end of the goal box is a pseudo-home cage that contains bedding similar to that found in the animal&#x2019;s home cage and is specific to each animal for the duration of maze testing. It has been demonstrated previously that this pseudo-home cage provides sufficient reward to motivate mice to learn to navigate the maze1. Here, we present the visualization of the Lashley III maze procedure in the context of evaluating age-related differences in learning and memory in mice along with a comparison of learning behavior in two different background strains of mice. We hope that other investigators interested in evaluating the effects of aging or stress vulnerability in mice will consider this maze an attractive alternative to behavioral tests that involve more stressful learning tasks and/or visual cues.

The Lashley III maze is a route-learning task that does not rely on aversive stimuli or visual cues. It is thus a highly attractive option for evaluating learning and memory, especially in aging mice or otherwise where stress is a consideration.

Many behavior tests designed to assess learning and memory in rodents, particularly mice, rely on visual cues, food and/or water deprivation, or other ...

Quantifying Cognitive Decrements Caused by Cranial Radiotherapy

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For many patients afflicted with CNS and non-CNS malignancies, radiotherapy and chemotherapy offer the best options for disease control. These treatments however come at a cost, and nearly all cancer survivors (~11 million in the US alone as of 2006) incur some risk for developing cognitive dysfunction, with the most severe cases found in patients subjected to cranial radiotherapy (~200,000/yr) for the control of primary and metastatic brain tumors1. Particularly problematic are pediatric cases, whose long-term survival plagued with marked cognitive decrements results in significant socioeconomic burdens2. To date, there are still no satisfactory solutions to this significant clinical problem.
We have addressed this serious health concern using transplanted stem cells to combat radiation-induced cognitive decline in athymic rats subjected to cranial irradiation3. Details of the stereotaxic irradiation and the in vitro culturing and transplantation of human neural stem cells (hNSCs) can be found in our companion paper (Acharya et al., JoVE reference). Following irradiation and transplantation surgery, rats are then assessed for changes in cognition, grafted cell survival and expression of differentiation-specific markers 1 and 4-months after irradiation. To critically evaluate the success or failure of any potential intervention designed to ameliorate radiation-induced cognitive sequelae, a rigorous series of quantitative cognitive tasks must be performed. To accomplish this, we subject our animals to a suite of cognitive testing paradigms including novel place recognition, water maze, elevated plus maze and fear conditioning, in order to quantify hippocampal and non-hippocampal learning and memory. We have demonstrated the utility of these tests for quantifying specific types of cognitive decrements in irradiated animals, and used them to show that animals engrafted with hNSCs exhibit significant improvements in cognitive function3.
The cognitive benefits derived from engrafted human stem cells suggest that similar strategies may one day provide much needed clinical recourse to cancer survivors suffering from impaired cognition. Accordingly, we have provided written and visual documentation of the critical steps used in our cognitive testing paradigms to facilitate the translation of our promising results into the clinic.

Cognitive impairment resulting from the radiotherapeutic management of brain tumors represents a clinically intractable condition that adversely impacts quality of life. The capability to critically evaluate potential interventions for ameliorating radiation-induced cognitive decrements ultimately depends on the capability to undertake rigorous quantitative assessments of cognition.

With the exception of survival, cognitive impairment stemming from the clinical management of cancer is a major factor dictating therapeutic outcome. For ...

Medicine

Morris Water Maze Test: Optimization for Mouse Strain and Testing Environment

The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, including neurocognitive disorders such as Alzheimer&#8217;s disease. However, the background strain of the mouse model used can have a substantial effect on the observed behavioral phenotype, with some strains exhibiting superior learning ability relative to others. To ensure differences between transgene negative and transgene positive mice can be detected, identification of a training procedure sensitive to the background strain is essential. Failure to tailor the MWM protocol to the background strain of the mouse model may lead to under- or over- training, thereby masking group differences in probe trials. Here, a MWM protocol tailored for use with the F1 FVB/N x 129S6 background is described. This is a frequently used background strain to study the age-dependent effects of mutant P301L tau (rTg(TauP301L)4510 mice) on the memory deficits associated with Alzheimer&#8217;s disease. Also described is a strategy to re-optimize, as dictated by the particular testing environment utilized.

This manuscript describes a Morris water maze (MWM) protocol tailored for use with a commonly used mouse model of Alzheimer's disease. The MWM is widely used in transgenic mouse models. Implementation of a procedure sensitive to the background strain of the mouse model is essential for detecting group differences.

 The Morris water maze (MWM) is a commonly used task to assess hippocampal-dependent spatial learning and memory in transgenic mouse models of disease, ...

Morris Water Maze Experiment

The Morris water maze is widely used to study spatial memory and learning. Animals are placed in a pool of water that is colored opaque with powdered non-fat milk or non-toxic tempera paint, where they must swim to a hidden escape platform. Because they are in opaque water, the animals cannot see the platform, and cannot rely on scent to find the escape route. Instead, they must rely on external/extra-maze cues. As the animals become more familiar with the task, they are able to find the platform more quickly. Developed by Richard G. Morris in 1984, this paradigm has become one of the "gold standards" of behavioral neuroscience.

The Morris water maze is a well-accepted tool used to document the involvement of the hippocampus in a behavioral task.

The Morris water maze is widely used to study spatial memory and learning. Animals are placed in a pool of water that is colored opaque with powdered ...

Biology

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and validated for rat use. The Radial Water Tread (RWT) maze test of spatial memory (designed specifically for mice and requiring no swimming) has been shown previously to successfully distinguish between controlled cortical impact-induced TBI mice and sham controls. Here, a detailed protocol for this task is presented. The RWT maze capitalizes on the natural tendency of mice to avoid open areas in favor of hugging the sides of an apparatus (thigmotaxis). The walls of the maze are lined with nine escape holes placed above the floor of the apparatus, and mice are trained to use visual cues to locate the escape hole that leads out of the maze. The maze is filled with an inch of cold water, sufficient to motivate escape but not deep enough to require that the mouse swim. The acquisition period takes only four training days, with a test of memory retention on day five and a long-term memory test on day 12. The results reported here suggest that the RWT maze is a feasible alternative to rat-validated, swimming-based cognitive tests in the assessment of spatial memory deficits in mouse models of TBI.

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham controls.

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

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 ...

Motor and Hippocampal Dependent Spatial Learning and Reference Memory Assessment in a Transgenic Rat Model of Alzheimer's Disease with Stroke

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk factor for AD, stroke has also been implicated as a risk factor and an exacerbating factor. The co-morbidity of stroke and AD results in worsened stroke-related motor control and AD-related cognitive deficits when compared to each condition alone. To model the combined condition of stroke and AD, a novel transgenic rat model of AD, with a mutated form of amyloid precursor protein (a key protein involved in the development of AD) incorporated into its DNA, is given a small unilateral striatal stroke.
For a model with the combination of both stroke and AD, behavioral tests that assess stroke-related motor control, locomotion and AD-related cognitive function must be implemented. The cylinder task involves a cost-efficient, multipurpose apparatus that assesses spontaneous forelimb motor use. In this task, a rat is placed in a cylindrical apparatus, where the rat will spontaneously rear and contact the wall of the cylinder with its forelimbs. These contacts are considered forelimb motor use and quantified during video analysis after testing. Another cost-efficient motor task implemented is the beam-walk task, which assesses forelimb control, hindlimb control and locomotion. This task involves a rat walking across a wooden beam allowing for the assessment of limb motor control through analysis of forelimb slips, hindlimb slips and falls. Assessment of learning and memory is completed with Morris water maze for this behavioral paradigm. The protocol starts with spatial learning, whereby the rat locates a stationary hidden platform. After spatial learning, the platform is removed and both short-term and long-term spatial reference memory is assessed. All three of these tasks are sensitive to behavioral differences and completed within 28 days for this model, making this paradigm time-efficient and cost-efficient.

To investigate the co-morbid Alzheimer&#39;s disease (AD) and stroke condition in a novel model, three behavior tasks are described that assess both motor control and cognitive behaviors. These tasks include the beam-walk task, cylinder task and Morris water maze.

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk ...

A Needle Model for Vascular Cognitive Impairment Studies in Mice

A Mouse Model for Vascular Cognitive Impairment and Dementia Based on Needle-guided Asymmetric Bilateral Common Carotid Artery Stenosis

Vascular cognitive impairment and dementia (VCID) results from vascular brain injury. Given VCID&#39;s high incidence, which is expected to continue rising as the population ages, it is critical to establish a robust animal model for the disease. This paper presents a novel method of creating a mouse model of VCID that is based on asymmetric bilateral common carotid artery stenosis, which mimics human chronic cerebral hypoperfusion caused by carotid atherosclerosis.
Briefly, common carotid arteries (CCAs) are ligated to different gauge needles (32 G for the right CCA and 34 G for the left CCA) using 7-0 silk sutures followed by immediate needle removal. The remaining suture rings cause persistent blood flow reduction and long-term cognitive impairment associated with white matter injury, microinfarcts, and reactive gliosis, thus closely mimicking the pathogenesis of VCID. Importantly, in this needle model, the clinical representations do not revert with time, providing reliable long-term cognitive impairment. Moreover, the survival rate 24 weeks post surgery was 81.6%, which is higher compared to the other established models of VCID with a similar level of blood flow reduction.
Additional advantages include low material cost and compatibility with MRI to monitor brain injury in live animals since no metal is implanted. The main challenge in employing the needle model of VCID is the requirement for developing advanced surgical skills since mouse CCAs are less than 0.6 mm in diameter and are very fragile. High-quality visual representation of the surgery will thus help researchers to master this technique and advance our understanding of VCID, potentially leading to the development of novel therapeutic modalities to decrease the devastating cognitive decline associated with VCID.

The needle method of asymmetric bilateral common carotid artery stenosis is proposed to create a mouse model for vascular cognitive impairment and dementia. It results in longer-term outcomes compared to previously established models and is compatible with live MRI. Visual representation demonstrating the procedure provides guidance for mastering the surgery.

Vascular cognitive impairment and dementia (VCID) results from vascular brain injury. Given VCID&#39;s high incidence, which is expected to continue ...

The Morris Water Maze

The Morris water maze is one of the most widely used behavioral tests for studying spatial learning and memory. In the initial phases of this task, rodents must swim to a platform to escape from a pool of water. The platform is then hidden under the water&#8217;s surface, so that the animal is required to remember it&#8217;s location in order to escape.This simple yet powerful maze design can be used to assay cognitive function, study animal models of neurodegenerative disease, and test potential drug therapies.
This video provides an introduction to the Morris water maze and the principles surrounding its use, including a discussion of the different types of memory tested in the maze, important points to consider when designing and conducting this experiment, and the procedures for setup and running of the test. Several applications of the maze are examined, such as investigating how radiation treatment may lead to memory impairment. Finally, other types of water mazes, such as the 8-arm radial maze, are introduced to show how this paradigm can be adapted to engage different types of memory.

The Morris water maze is one of the most widely used behavioral tests for studying spatial learning and memory. In the initial phases of this task, ...