This work was supported by the University of Strasbourg and Neurex&#8212;Neuroscience Upper Rhine Network (post-doc fellowship to RP) and by BrainLinks-BrainTools Cluster of Excellence funded by the German Research Foundation (DFG, grant number EXC 1086). We thank Nadja Martini for expert technical assistance.

1. General Considerations
This protocol is approved by the Animal Care and Use Committee of University Hospital Freiburg (same for Strasbourg). Visual acuity is necessary for performance in tests of spatial learning. Rodents with impaired visual systems are thus not suitable. Also, lighting must be sufficient in order for the rats to see the different cues located on the surrounding walls. It is useful to utilize basic-shaped (square, circle, triangle) but well-contrasted cues (e.g., black-paint...

<ol>
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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rats+with+fimbria-fornix+lesions+display+a+place+response+in+a+swimming+pool:+a+dissociation+between+getting+there+and+knowing+where.">Rats with fimbria-fornix lesions display a place response in a swimming pool: a dissociation between getting there and knowing where.</a> J Neurosci. 15 (8), 5779-5788 (1995).</li><li>Staay, F. J., Raaijmakers, W. G., Lammers, A. J., Tonnaer, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+fimbria+lesions+impair+acquisition+of+working+and+reference+memory+of+rats+in+a+complex+spatial+discrimination+task.">Selective fimbria lesions impair acquisition of working and reference memory of rats in a complex spatial discrimination task.</a> Behav Brain Res. 32 (2), 151-161 (1989).</li><li>Olton, R. S., Samuelson, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remembrance+of+places+past:+Spatial+memory+in+rats.">Remembrance of places past: Spatial memory in rats.</a> J Exp Psych. 2 (2), 97-116 (1976).</li><li>Pol-Bodetto, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+double-H+maze+test,+a+novel,+simple,+water-escape+memory+task:+acquisition,+recall+of+recent+and+remote+memory,+and+effects+of+systemic+muscarinic+or+NMDA+receptor+blockade+during+training.">The double-H maze test, a novel, simple, water-escape memory task: acquisition, recall of recent and remote memory, and effects of systemic muscarinic or NMDA receptor blockade during training.</a> Behav Brain Res. 218 (1), 138-151 (2011).</li><li>Cassel, R., Kelche, C., Lecourtier, L., Cassel, J. -. 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The authors have nothing to disclose.

Comments on Study Design and Analysis
Since its conception, the double-H maze has been utilized in a number of behavioral experiments in rats, which collectively were designed to study egocentric and/or allocentric responses in rats under normal24,25 and altered26-29 brain states. The latter studies include striatal deep-brain stimulation (DBS)26, animal models of neurological disorders27,28, as well as bilateral deactivations of various cortico-hippoca...

In animals, learning is principally mediated by the hippocampal- and striatal-based memory systems1,2, which play central roles regarding place- and procedural-memory, respectively. The relationship between these two systems is complex, and they are known to interact with each other in cooperative or competitive manners1,3. In addition, studies have shown that the influence of either of these memory systems on animal behavior can increase following the absence or damage of the other system4-7. Both of these systems are connected to the prefrontal cortex via the thalamus.
Numerous neurological disorders and neurodegenerative diseases can affect spatial cognition in humans, which rely on the interplay between procedural and declarative memory systems. Examples include Parkinson&#8217;s disease (PD), Huntington&#8217;s disease (HD)8-10, Alzheimer&#8217;s disease (AD)11-14, as well as amyotrophic lateral sclerosis (ALS)15. Animal models, which are relevant to these disorders can be induced through various drug treatments which block certain receptors16, as well as through targeted lesions. When such animals are used with spatial memory tasks, a valuable insight can be gained into the underlying mechanisms related to these disorders, as well as to various treatment options.
There are many different types of spatial memory tasks in rodents, which collectively are designed to assess specific aspects of learning and memory, as well as the effects of potential treatments for various disorders17,18. These tasks can be distinguished by the number of goals and pathways, the degree of behavioral flexibility in solving the task, the memory duration or delay, as well as the choice of strategy used in solving the task. A good performance may be acquired based on external cues or landmarks which are used to orientate the animal towards the goal (an allocentric or place strategy). Alternatively, a rodent may develop a strategy which is based on bodily direction and cues with regards to the direction to move in (an egocentric or procedural strategy), e.g., if a rat knows that the goal is one left turn followed by one right turn, then there is little need for an allocentric or place strategy. Maze tasks often differ based on the degree of flexibility offered to the rodent in solving them. For instance, in the Morris Water Maze, a dry version of the latter (e.g., 19) or the Barnes maze (e.g., 20), there are potentially infinite routes the rat can take to reach the goal. In the Morris Water Maze, for example, the location of the goal may be learned based on external landmarks or cues (allocentric strategy), or by simply swimming in circles towards the center until the platform is found (egocentric strategy)21. Certain tasks have multiple goals and a high degree of flexibility, such as the cone-field task22 or Olton&#8217;s radial maze23. At the other end of the scale are tasks, which offer limited flexibility in reaching the goal, e.g., the Stone maze, or the alternating version of the T-maze. These tasks provide only one correct way of reaching the goal and facilitate the emergence of cognitive routines that are principally governed by the striatal-based procedural memory system.
The double-H maze is a novel spatial memory testing device, which was designed to allow the experimenter to direct the type of strategy that is learned by rodents in solving the task24. Consisting of three parallel run arms intersected by a perpendicular central alleyway, the double-H maze is a water-escape task in which rodents learn to reach an escape platform that is immersed in one of the maze locations. During training, a procedural strategy can be developed by maintaining the same start and goal locations throughout. Alternatively, an allocentric strategy may be developed by alternating the starting location in a random order, thus requiring the rat to learn the location of the hidden platform based on environmental cues as it has to do in a water maze. This overcomes an obstacle present in many different maze tasks, in which the experimenter otherwise has little control over the type of strategy that rodents utilize. This is important when considering that the effects of certain cognition-enhancing drug candidates rely on the hippocampal-based place-memory system, thus the emergence of cognitive routines or procedures may confound the interpretation of the behavioral observations when animals, for example switch from allocentric to procedural memory during the course of training. Similarly, it may be desirable to assess the effects of drugs and treatments on procedural memory, without the influence of allocentric place-based memory. Finally, this device can be utilized to study the cooperative or competitive interactions between these memory systems, and the conditions under which rodents may switch from one system to another.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Rats or Mice</td>
			<td></td>
			<td>Charles River</td>
			<td></td>
		</tr>
		<tr>
			<td>Towels for drying</td>
			<td></td>
			<td>University Hospital</td>
			<td>1 / animal</td>
		</tr>
		<tr>
			<td>Water</td>
			<td></td>
			<td></td>
			<td>~200 L / day</td>
		</tr>
		<tr>
			<td>Skim milk powder</td>
			<td></td>
			<td>Grocery store</td>
			<td>250 g / 200 L water</td>
		</tr>
		<tr>
			<td>Garden Hose</td>
			<td></td>
			<td>Hardware store</td>
			<td></td>
		</tr>
		<tr>
			<td>Drying rack for towels</td>
			<td></td>
			<td>Hardware store</td>
			<td></td>
		</tr>
		<tr>
			<td>Kinect camera</td>
			<td></td>
			<td>Kinect</td>
			<td></td>
		</tr>
		<tr>
			<td>PC computer</td>
			<td></td>
			<td>any</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>[header]</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Double H Maze, (plexiglass) (Custom-Built)</td>
		</tr>
		<tr>
			<td>External lateral walls, 1600 &#215; 350 &#215; 6 mm</td>
			<td></td>
			<td align="right">2</td>
			<td></td>
		</tr>
		<tr>
			<td>Internal lateral walls, 706 &#215; 350 &#215; 6 mm</td>
			<td></td>
			<td align="right">8</td>
			<td></td>
		</tr>
		<tr>
			<td>Central corridor panels, 500 &#215; 350 &#215; 6 mm</td>
			<td></td>
			<td align="right">4</td>
			<td></td>
		</tr>
		<tr>
			<td>Arm extremities, 188 &#215; 350 &#215; 6 mm</td>
			<td></td>
			<td align="right">6</td>
			<td></td>
		</tr>
		<tr>
			<td>Guillotine doors, 187 &#215; 350 &#215; 6 mm</td>
			<td></td>
			<td align="right">3</td>
			<td></td>
		</tr>
		<tr>
			<td>Extremity covers, 200 &#215; 250 mm</td>
			<td></td>
			<td align="right">6</td>
			<td></td>
		</tr>
		<tr>
			<td>Crossbars, 200 &#215; 40 mm</td>
			<td></td>
			<td align="right">6</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>[header]</td>
		</tr>
		<tr>
			<td>Double-H Maze Platform (to be ballasted)</td>
			<td></td>
			<td></td>
			<td>Double-H Maze Platform (to be ballasted) (Custom-Built)</td>
		</tr>
		<tr>
			<td>Metal platform, 100 mm diameter &#215; 150 mm</td>
			<td></td>
			<td align="right">2</td>
			<td></td>
		</tr>
		<tr>
			<td>Platform cover, 100 mm diameter &#215; 6mm</td>
			<td></td>
			<td align="right">2</td>
			<td></td>
		</tr>
	</tbody>
</table>

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.

Egocentric Learning Strategy
A study was carried out to determine whether the chosen memory strategy in rats changes based on alterations of their perspective of external environmental cues, following an egocentric-learning paradigm25. Rats were trained over 4 days (4 trials/day) to reach a goal arm located at NE, and were subsequently tested on the fifth day using a misleading probe trial, in which the start arm was either moved 60 cm to the left (i.e., NE start for animals r...

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

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 ...

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12.4K Views.  University Hospital Freiburg. 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.