Thank you to all the participants that have helped develop this methodology over different iterations of the task. Thank you also to all the students who helped by running the different iterations over the years: Natasha Dubes, Emma Denning, Victoria Bellhouse, Stephen Holland, Melissa Anderson, Katie Shaw, Sarah Morgan, Karla Butterworth, Michael Craig, Lauren Wray, Olivia Sanderson, Daniel Lai, Rajameenakshi Boopathy and Chun Kit Ho. This research was funded by Newcastle University's contributions to student research projects.

This protocol was approved by Newcastle University&#39;s Faculty of Medical Sciences Ethics Committee (approval number 515_1).
1. Preparation for the study
<ol>
	<li>Before running the study, gather 20 small, easily identified/described objects, e.g., 
	A tea light 
	A toy digger 
	A toy frog 
	A clothing hook 
	A spoon 
	A set of keys 
	A button 
	A lip balm 
	A toy snowman 
	A bottle cap 
	A die 
	A lighter 
	A comb 
	A wrist band 
	A padlock 
	A butterfly pin 
	A ruler 
	A guitar plectrum 
	A battery 
	A USB stick</li>
	<li>Randomly select 8 of these objects for the first session, and 8 for the second session. There should be 4 extra objects that are not used in either session. Once the objects have been selected, use the same objects in the same locations and in the same sessions for all participants.</li>
	<li>Make photographs of the 16 objects to be used. Make two sheets of these photographs: the first sheet for the first session (8 objects) and the second sheet for the second session (8 objects).
	<ol>
		<li>Put the images of the objects on the sheet in the order the participants need to find them. Number the images to avoid any confusion. Figure 1 shows an example.</li>
	</ol>
	</li>
</ol>
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55646/55646fig1.jpg" /> 
Figure 1: Object identity sheets. These are the sheets that are placed next to the pile of 20 objects. The left-hand sheet is placed there in phase 1 and the right-hand sheet in phase 2. Participants are supposed to pick up the objects from the pile in the order indicated on the sheet. Note that in phase 2, all the objects from phase 1 have been collected up and added to the pile again, so that during phase 2, participants again search through a pile of 20 objects to start with. <a href="http://cloudfront.jove.com/files/ftp_upload/55646/55646fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<ol start="4">
	<li>Identify 16 hiding locations around the room. Ideally, the room is an office with many things in it. The locations should be unambiguously describable by a person not actually in the room. Also, try and make the locations not purely associated with a clear object in the room, as this reduces spatial memory to object-object associations.
	<ol>
		<li>Assign each object to one of the hiding locations, making sure that the locations used in the first and second session are randomly interspersed.</li>
	</ol>
	</li>
	<li>Make sure that the lay-out of the room will remain the same for the duration of the study. In multi-use rooms, make sure that the room can be put back in the same way before each participant.</li>
</ol>
2. Session 1
<ol>
	<li>Before the participant arrives, put the 20 objects in a pile in a fixed location in the testing room, with the object picture sheet for session 1 next to it. The reason for having participants find the objects in a pile using the picture sheets is to force them to pay attention to the identity of the objects.</li>
	<li>When the participant arrives, first give them an information leaflet explaining the study, talk them through the study, and have them read and sign the consent forms.</li>
	<li>Take the participants to the door of the testing room and give instructions about what they will need to do in the room. The instructions can be modified to either induce intentional memorization or incidental memorization.
	<ol>
		<li>For intentional memorization, read these instructions to the participant: 
		&#34;The purpose of this task is for you to hide some objects in a room and you will be asked to remember them later. You will be presented with a pile of objects on the desk. Next to the pile you will find a sheet with pictures of the 8 objects that you must hide during this task. At the bottom left of each picture there is a number indicating the order in which you must hide these objects. 
		You may only pick up and hide one object at a time. I will point to the location where you should hide each object. Once you enter the room you should start counting seconds out loud and continue doing that until you leave the room. 
		Later in this study we will repeat this with different objects and different locations. After this you will be asked to remember what objects you hid, where you hid them and on which occasion. If you have any questions, please ask them now because you will not be able to once we enter the room. This is not a timed task, so please take as much time as you need.&#34;</li>
		<li>For incidental memorization, read these instructions to the participant: 
		&#34;The purpose of this task is to test your multi-tasking abilities. You will need to count seconds out loud, without slowing down or skipping numbers, while I try to distract you with objects to look for and place in different places around the room. I will record your voice for later analysis of your counting. 
		For the distractor, you will be presented with a pile of objects on the desk. Next to the pile you will find a sheet with pictures of the 8 objects that you must hide during this task. At the bottom left of each picture there is a number indicating the order in which you must hide these objects. You may only pick up and hide one object at a time. I will point to the location where you should hide each object. Once you enter the room you should start counting seconds out loud and continue doing that until you leave the room. 
		Later in this study we will repeat this to test whether you get better with practice. If you have any questions, please ask them now because you will not be able to once we enter the room. This is not a timed task, so please take as much time as you need, but don&#39;t forget to keep the count going at a steady rate.&#34;</li>
	</ol>
	</li>
	<li>Take the participant into the room and show the pile of objects. Have the participant find and take the first object. Indicate where the object needs to be placed, and have the participant place the object there.
	<ol>
		<li>Then have the participant go back for the next object, and so on for all 8 objects on the picture sheet. The timing is&#160;determined by the speed at which the participant finds and hides the objects. This typically takes no more than 2 minutes.</li>
	</ol>
	</li>
	<li>At the end of the session, take the participant out of the room.</li>
</ol>
3. First break
<ol>
	<li>Between session 1 and session 2, leave approximately 2 hours. Ask participants to do different tasks during this interval, or ask them to go away (e.g. to have lunch) and to come back at the appointed time.</li>
	<li>Before the participant returns, remove all the hidden objects from the room and put them back in the pile.</li>
	<li>Replace the picture sheet with the picture sheet for session 2.</li>
</ol>
4. Session 2
<ol>
	<li>Remind the participant of the instructions given before session 1.</li>
	<li>Take the participant into the room, and repeat section 2.4, but now with different objects and locations. Again, the timing is determined by the participant.</li>
	<li>Take the participant back out of the room.</li>
</ol>
5. Second break
<ol>
	<li>Again instruct the participant to come back 2 h later or run another battery of tests during this interval.</li>
	<li>Remove all objects from their hiding places. The room will not be needed again for this participant.</li>
</ol>
6. Session 3
<ol>
	<li>If the participants had been instructed for incidental memorization, debrief them now and tell them the real purpose of the task. Check the effectiveness of the deceit by asking the participants if they suspected the task was a memory task.</li>
	<li>Ask them to freely recall which objects they had hidden in which locations and on which of the two occasions. Ask them to write this all down in the order in which they remember them. Encourage them to recall any information they can, including incomplete information (like remembering an object, but not its location, etc.).
	<ol>
		<li>Give the participants all the time they need to recall as much as they can. Allow them to draw a map or diagram if they so wish.</li>
	</ol>
	</li>
	<li>After they have recalled all the information they can, ask them to complete the vividness scale and the task contemplation scale (Figure 2).</li>
</ol>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55646/55646fig2.jpg" /> 
Figure 2: Subjective experience reporting. After reporting on their memory for objects, locations and phases, the participants are asked to complete these two scales, which report on the subjective experience of their memories (top scale) and on how much they actually rehearsed the material after each hiding phase. <a href="http://cloudfront.jove.com/files/ftp_upload/55646/55646fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<ol start="4">
	<li>Debrief participants further and send away.</li>
</ol>
7. Data extraction and analysis
<ol>
	<li>For the scoring sheet, create a list of all 16 objects, and another list of all 16 locations.
	<ol>
		<li>For each object and location, determine whether it was remembered in combination with the correct session and location/object (respectively; What-Where-When)), whether in combination only with the correct session (only What-When or only Where-When) or only with the correct location/object (respectively; only What-Where), or whether it was recalled correctly, but without any correct combination (only What and only Where). The number of correctly recalled What-Where-When combinations and What-Where combinations should match for both lists.</li>
	</ol>
	</li>
	<li>Analyze the full What-Where-When combinations first. Because there is a total of 16 combinations to be recalled, treat the number of actually correctly recalled combinations as coming from a binomial distribution (16 &#39;yes-no&#39; answers), which should be analyzed with a Generalized Linear Model, using a binomial distribution with log-link function.</li>
	<li>Then analyze the 3 incomplete combinations. To do this using the same analysis, analyze the total number of incompletely recalled combinations of each type out of the remaining number of combinations that were not correctly recalled as a complete what-where-when combination. For example, if a participant recalled 5 correct what-where-when combinations, then analyze incomplete combinations out of (16-5=) 11 possible incomplete combinations.</li>
	<li>Analyze the number of objects or locations recalled without any combination in an analogous way out of the remaining combinations that have not yet been accounted for.</li>
</ol>

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The authors have nothing to disclose.

The data show that performance on other tasks which are supposed to measure episodic memory predicts performance on the Real-World What-Where-When memory task as well. However, these correlations are likely to represent a shared subset of cognitive abilities used by the different tasks. The Real-World What-Where-When memory task has the advantage over these other tasks in that it tests people&#39;s memory for two actual events which happened in a real spatio-temporal context. Unlike asking people about events from their ...

Episodic memory is memory for unique events from one&#39;s own past that are experienced as a reliving of the original event (mental time travel)1,2. It is also one of the first types of memory to be affected in the early stages of many forms of dementia3,4. The medial temporal lobe, and more specifically the hippocampus, is believed to be an important structure in the processing of episodic memories5, and therefore any conditions that affect hippocampal function, like aging and many mood disorders, are also believed to affect episodic memory function. As such, episodic memory function can be a useful biomarker for a range of neurological and psychiatric conditions6.
Methods for quantifying episodic memory, however, are still less than ideal. Real-world everyday episodic memories are integrative memories of unique events in their spatiotemporal context7, usually incidentally encoded4. The two most common methods used in both the clinic and in academic research are word list learning8 and recounting a story from one&#39;s own past3. Both methods have advantages and disadvantages. The advantage of the word lists over the story approach is that the assessor knows exactly what the right answers are. This is difficult to assess with spontaneous stories from the participant/patient&#39;s past, since often no objective evidence is available and even accounts from family members may have incorrect details in them. The advantage of the stories is that they actually assess the typical content and structure of episodic memories: events in spatiotemporal context, with information about What happened, Where and When bound together7. Word lists do not require any context to be recalled at all, and are often rehearsed several times (e.g. the Rey Auditory Verbal Learning Task).
Recently, several attempts have been made to construct episodic memory tasks that combine the strengths of the two classical tests while minimizing the drawbacks9,10,11,12,13,14,15,16,17,18,19,20,21,22. The current protocol is the most recent version of a What-Where-When episodic memory test which has been developed at Newcastle University10,16,22. The concept is based on the work with non-human animals, started by Clayton and Dickinson23, and adapted for work with a range of other species24,25,26,27,28, some of which have confirmed the sensitivity of this paradigm to medial temporal lobe damage29. It is one of several attempts at incorporating a What-Where-When framework in episodic memory testing with adult humans18,20,30,31, but the only one to be performed in a real environment, without the use of computers, making it easy for participants/patients to engage with and low-cost to carry out.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>These materials are just examples. Any 20 easy-to-name small objects will do.</td>
		</tr>
		<tr>
			<td>- A tea light</td>
		</tr>
		<tr>
			<td>- A toy digger</td>
		</tr>
		<tr>
			<td>- A toy frog</td>
		</tr>
		<tr>
			<td>- A clothing hook</td>
		</tr>
		<tr>
			<td>- A spoon</td>
		</tr>
		<tr>
			<td>- A set of keys</td>
		</tr>
		<tr>
			<td>- A button</td>
		</tr>
		<tr>
			<td>- A lip balm</td>
		</tr>
		<tr>
			<td>- A toy snowman</td>
		</tr>
		<tr>
			<td>- A bottle cap</td>
		</tr>
		<tr>
			<td>- A die</td>
		</tr>
		<tr>
			<td>- A lighter</td>
		</tr>
		<tr>
			<td>- A comb</td>
		</tr>
		<tr>
			<td>- A wrist band</td>
		</tr>
		<tr>
			<td>- A padlock</td>
		</tr>
		<tr>
			<td>- A butterfly pin</td>
		</tr>
		<tr>
			<td>- A ruler</td>
		</tr>
		<tr>
			<td>- A guitar plectrum</td>
		</tr>
		<tr>
			<td>- A battery</td>
		</tr>
		<tr>
			<td>- A USB stick</td>
		</tr>
	</tbody>
</table>

a real-world what-where-when memory test

Episodic memory is a complex memory system which allows recall and mental re-experience of previous episodes from one&#39;s own life. Real-life episodic memories are about events in their spatiotemporal context and are typically visuospatial, rather than verbal. Yet often, tests of episodic memory use verbal material to be recalled (word lists, stories). The Real-World What-Where-When memory test requires participants to hide a total of 16 different objects in 16 different locations over two temporal occasions, 2 h apart. Another two hours later, they are then asked to recall which objects (What) they had hidden in which locations (Where) and on which of the two occasions (When). In addition to counting the number of correctly recalled complete what-where-when combinations, this task can also be used to test real-world spatial memory and object memory. This task is sensitive to normal cognitive aging, and correlates well with performance on other episodic memory tasks, while at the same time providing more ecological validity and being cheap and easy to run.

Older people (65+) remember fewer complete What-Where-When combinations than do younger people (18-25; &#935;2(1) = 9.5; p = 0.002; Figure 3). Note that although as a group, older people perform worse than younger people, there are some older people who perform as well or better than some young people. This variation may be informative if it is predictive of other conditions.
One can also...

Watch this Scientific Journal Video about A Real-world What-Where-When Memory Test at JoVE.com

A Real-world What-Where-When Memory Test

The Real-World What-Where-When memory test is a novel episodic memory test, in which participants need to recall which objects have been hidden in which locations on which of two distinct occasions. It is easy to run and is sensitive to normal cognitive aging.

A Real-world What-Where-When Memory Test

Episodic memory is a complex memory system which allows recall and mental re-experience of previous episodes from one&#39;s own life. Real-life episodic ...

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11.1K Views.  Newcastle University. The Real-World What-Where-When memory test is a novel episodic memory test, in which participants need to recall which objects have been hidden in which locations on which of two distinct occasions. It is easy to run and is sensitive to normal cognitive aging.

Video: A Real-world What-Where-When Memory Test

Disrupting Reconsolidation of Fear Memory in Humans by a Noradrenergic &#946;-Blocker

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive days. On day 1 - the fear acquisition phase, healthy participants are exposed to a series of picture presentations. One picture stimulus (CS1+) is repeatedly paired with an aversive electric stimulus (US), resulting in the acquisition of a fear association, whereas another picture stimulus (CS2-) is never followed by an US. On day 2 - the memory reactivation phase, the participants are re-exposed to the conditioned stimulus without the US (CS1-), which typically triggers a conditioned fear response. After the memory reactivation we administer an oral dose of 40 mg of propranolol HCl, a &#946;-adrenergic receptor antagonist that indirectly targets the protein synthesis required for reconsolidation by inhibiting the noradrenaline-stimulated CREB phosphorylation. On day 3 - the test phase, the participants are again exposed to the unreinforced conditioned stimuli (CS1- and CS2-) in order to measure the fear-reducing effect of the manipulation. This retention test is followed by an extinction procedure and the presentation of situational triggers to test for the return of fear. Potentiation of the eye blink startle reflex is measured as an index for conditioned fear responding. Declarative knowledge of the fear association is measured through online US expectancy ratings during each CS presentation. In contrast to extinction learning, disrupting reconsolidation targets the original fear memory thereby preventing the return of fear. Although the clinical applications are still in their infancy, disrupting reconsolidation of fear memory seems to be a promising new technique with the prospect to persistently dampen the expression of fear memory in patients suffering from anxiety disorders and other psychiatric disorders.

Disrupting reconsolidation is a promising approach to dampen the behavioral expression of fear memory in patients with anxiety disorders or posttraumatic stress disorder. In a series of human fear conditioning studies we showed that disrupting reconsolidation by the noradrenergic &#946;-blocker propranolol is very effective in erasing conditioned fear responding.

The basic design used in our human fear-conditioning studies on disrupting reconsolidation includes testing over different phases across three consecutive ...

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 ...

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, ...

Using Fiberless, Wearable fNIRS to Monitor Brain Activity in Real-world Cognitive Tasks

Functional Near Infrared Spectroscopy (fNIRS) is a neuroimaging technique that uses near-infrared light to monitor brain activity. Based on neurovascular coupling, fNIRS is able to measure the haemoglobin concentration changes secondary to neuronal activity. Compared to other neuroimaging techniques, fNIRS represents a good compromise in terms of spatial and temporal resolution. Moreover, it is portable, lightweight, less sensitive to motion artifacts and does not impose significant physical restraints. It is therefore appropriate to monitor a wide range of cognitive tasks (e.g., auditory, gait analysis, social interaction) and different age populations (e.g., new-borns, adults, elderly people). The recent development of fiberless fNIRS devices has opened the way to new applications in neuroscience research. This represents a unique opportunity to study functional activity during real-world tests, which can be more sensitive and accurate in assessing cognitive function and dysfunction than lab-based tests. This study explored the use of fiberless fNIRS to monitor brain activity during a real-world prospective memory task. This protocol is performed outside the lab and brain haemoglobin concentration changes are continuously measured over the prefrontal cortex while the subject walks around in order to accomplish several different tasks.

Monitoring brain activity outside the lab without physical constraints presents methodological challenges. A fiberless, wearable functional Near Infrared Spectroscopy (fNIRS) system was used to measure brain activity during an ecological prospective memory task. It was demonstrated that this system could be used to monitor brain activity during non-lab based experiments.

Functional Near Infrared Spectroscopy (fNIRS) is a neuroimaging technique that uses near-infrared light to monitor brain activity. Based on neurovascular ...

The 4 Mountains Test: A Short Test of Spatial Memory with High Sensitivity for the Diagnosis of Pre-dementia Alzheimer's Disease

This protocol describes the administration of the 4 Mountains Test (4MT), a short test of spatial memory, in which memory for the topographical layout of four mountains within a computer-generated landscape is tested using a delayed match-to-sample paradigm. Allocentric spatial memory is assessed by altering the viewpoint, colors and textures between the initially presented and target images.
Allocentric spatial memory is a key function of the hippocampus, one of the earliest brain regions to be affected in Alzheimer&#39;s disease (AD) and impairment of hippocampal function predates the onset of dementia. It was hypothesized that performance on the 4MT would aid the diagnosis of predementia AD, which manifests clinically as Mild Cognitive Impairment (MCI).
The 4MT was applied to patients with MCI, stratified further based on cerebrospinal fluid (CSF) AD biomarker status (10 MCI biomarker positive, 9 MCI biomarker negative), and with mild AD dementia, as well as healthy controls. Comparator tests included tests of episodic memory and attention widely accepted as sensitive measures of early AD. Behavioral data were correlated with quantitative MRI measures of the hippocampus, precuneus and posterior cingulate gyrus.
4MT scores were significantly different between the two MCI groups (p = 0.001), with a test score of &#8804;8/15 associated with 100% sensitivity and 78% specificity for the classification of MCI with positive AD biomarkers, i.e., predementia AD. 4MT test scores correlated with hippocampal volume (r = 0.42) and cortical thickness of the precuneus (r = 0.55).
In conclusion, the 4MT is effective in identifying the early stages of AD. The short duration, easy application and scoring, and favorable psychometric properties of the 4MT fulfil the need for a simple but accurate diagnostic test for predementia AD.

This article describes the 4 Mountains Test (4MT), a hippocampus-dependent test of working allocentric spatial memory. The hippocampus is affected early in Alzheimer's disease (AD) and this article outlines the 4MT methodology and results of patient testing, which demonstrates the value of the 4MT in the diagnosis of pre-dementia AD.

This protocol describes the administration of the 4 Mountains Test (4MT), a short test of spatial memory, in which memory for the topographical layout of ...

Assessing Working Memory in Children: The Comprehensive Assessment Battery for Children &#8211; Working Memory (CABC-WM)

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of working memory in young school-age children. Working memory deficits have been identified in children with language-based learning disabilities, including dyslexia1,2 and language impairment3,4, but it is not clear whether these children exhibit deficits in subcomponents of working memory, such as visuospatial or phonological working memory. The CABC-WM is administered on a desktop computer with a touchscreen interface and was specifically developed to be engaging and motivating for children. Although the long-term goal of the CABC-WM is to provide individualized working memory profiles in children, the present study focuses on the initial success and utility of the CABC-WM for measuring central executive, visuospatial, phonological loop, and binding constructs in children with typical development. Immediate next steps are to administer the CABC-WM to children with specific language impairment, dyslexia, and comorbid specific language impairment and dyslexia.

Working memory predicts a significant amount of variance for a variety of cognitive tasks, including speaking, reading, and writing. However, few tools are available to assess working memory in children. We present an innovative, computer-based battery that comprehensively assesses different components of working memory in school-age children.

The Comprehensive Assessment Battery for Children - Working Memory (CABC-WM) is a computer-based battery designed to assess different components of ...

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors remains poorly understood. In Drosophila melanogaster, environmental cues, such as food availability, affect mating activity offering a tractable system to investigate the mechanisms modulating sexual behavior. In D. melanogaster, environmental cues are often sensed via the chemosensory gustatory and olfactory systems. Here, we present a method to test the effect of environmental chemical cues on mating behavior. The assay consists of a small mating arena containing food medium and a mating couple. The mating frequency for each couple is continuously monitored for 24 h. Here we present the applicability of this assay to test environmental compounds from an external source through a pressurized air system as well as manipulation of the environmental components directly in the mating arena. The use of a pressurized air system is especially useful to test the effect of very volatile compounds, while manipulating components directly in the mating arena can be of value to ascertain a compound's presence. This assay can be adapted to answer questions about the influence of genetic and environmental cues on mating behavior and fecundity as well as other male and female reproductive behaviors.

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

We demonstrate an assay to analyze the environmental and genetic cues that influence mating behavior in the fruit fly Drosophila melanogaster.

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

Novel Object Recognition Test for the Investigation of Learning and Memory in Mice

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is fairly simple and can be completed over 3 days: habituation day, training day, and testing day. During training, the mouse is allowed to explore 2 identical objects. On test day, one of the training objects is replaced with a novel object. Because mice have an innate preference for novelty, if the mouse recognizes the familiar object, it will spend most of its time at the novel object. Due to this innate preference, there is no need for positive or negative reinforcement or long training schedules. Additionally, the ORT can also be modified for numerous applications. The retention interval can be shortened to examine short-term memory, or lengthened to probe long-term memory. Pharmacological intervention can be used at various times prior to training, after training, or prior to recall to investigate different phases of learning (i.e., acquisition, early or late consolidation, or recall). Overall, the ORT is a relatively low-stress, efficient test for memory in mice, and is appropriate for the detection of neuropsychological changes following pharmacological, biological, or genetic manipulations.

The object recognition test (ORT) is a simple and efficient assay for evaluating learning and memory in mice. The methodology is described below.

The object recognition test (ORT) is a commonly used behavioral assay for the investigation of various aspects of learning and memory in mice. The ORT is ...

Reducing State Anxiety Using Working Memory Maintenance

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working Memory (WM) and the threat of shock paradigms. In the Sternberg WM paradigm, subjects are required to maintain a series of letters in the WM for a brief interval and respond by identifying whether the position of a given letter in the series matches a numerical prompt. In the threat of shock paradigm, subjects are exposed to alternating blocks where they are either at risk of receiving unpredictable presentations of a mild electric shock or are safe from the shock. Anxiety is probed throughout the safe and threat blocks using the acoustic startle reflex, which is potentiated under threat (Anxiety-Potentiated Startle (APS)). By conducting the Sternberg WM paradigm during the threat of shock and probing the startle response during either the WM maintenance interval or the intertrial interval, it is possible to determine the effect of WM maintenance on APS.

This protocol demonstrates how to measure anxiety-potentiated startle during the Sternberg Working Memory paradigm.

The purpose of this protocol is to explain how to examine the relationship between working memory processes and anxiety by combining the Sternberg Working ...

Olfactory Context Dependent Memory: Direct Presentation of Odorants

Information is retrieved more effectively when the retrieval occurs in the same context as that in which the information was first encoded. This is termed context dependent memory (CDM). One category of cues that have been shown to effectively produce CDM effects are odors. However, it is unclear what the boundary conditions of these CDM effects are. In particular, given that olfaction has been called an implicit sense, it is possible that odors are only effective mnemonic cues when they are presented in the background. This assertion seems even more likely given that previous research has shown odors to be poor cues during paired associate memory tests, where odors are in the focus of attention as mnemonic cues for other information. In order to determine whether odors are only effective contextual mnemonic cues when presented outside the central focus of an observer, an olfactory CDM experiment was performed in which odorants were presented directly, rather than ambiently. Direct presentation was accomplished with the aid of an olfactometer. The olfactometer not only allows for direct presentation of odorants, but provides other methodological benefits, including the allowance of trial by trial manipulations of odorant presentations and, relatedly, time-specific releases of odorants. The presence of the same odor during both encoding and retrieval enhanced memory performance, regardless of whether the odor was presented ambiently or directly. This finding can serve as a basis for future olfactory CDM research which can utilize the benefits of direct presentation.

The use of an olfactometer for directly presenting odorants opens exciting opportunities for researchers of olfactory memory. The current paper discusses issues related to this methodology as related to a previously published experiment on olfactory context dependent memory.

Information is retrieved more effectively when the retrieval occurs in the same context as that in which the information was first encoded. This is termed ...