We thank Reiko Ando for her help in photographing materials. This research was supported by Grant-in-Aid for Exploratory Research (JSPS KAKENHI Grant Number 16K15196).

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No conflicts of interest declared.

This paper describes the method using the automated home-cage monitoring system for long-term cognitive and behavioral assays in genetically modified AD models. The most critical step is the implantation of the transponder in the appropriate position. Before performing the implantation, ensure that the expiration date of the transponder has not passed. The second important point is to check the functioning of the system daily, especially as a minor problem can subsequently become a more serious one during the study (i.e., a stacked door, fallen-out transponder, bad electrical connections, etc.). Third, it is essential to be able to trouble shoot because many issues may occur throughout the experimental schedule.
This paper introduced a basic package of tasks for cognitive assessment. These tasks were produced by referring to the conventional behavioral tests, but they cannot perfectly mimic the conventional tests. For example, SRT tasks do not set choice-mode. Five choice serial choice task (5CSRTT), a model of SRT tasks, is normally conducted in a chamber with 9, 5 or 3 holes for the nosepoke to measure the accuracy16,17. Our team previously tried to implement a choice version of SRT, but the mice failed to learn the rule within relatively long time (2 weeks or more). The user of this system should be aware of the difference, and discuss data carefully.
Another issue is about the limitations for the repeated measurement. As mentioned in previous study18, the first one is that the repeated experiments cannot simply compare the effect of age. We observed that the data from the second PA test failed to replicate the memory impairment in AD models10. The performance of all genotype was much worse than that of the first PA test. This difference may come from age difference or habituation to the aversive stimulus (experience of previous PA task). The experimenters should mind the repeat effect and consider the order of the tasks. To overcome the habituation of aversive stimulus in the second PA test, it might be better to use stronger air-puff stimulus or to add some novel environmental stimulus such as different types of bedding or sound19.
Various researchers have been developing new methods and protocols for the automated home-cage monitoring studies20,21,22,23,24,25,26, and supporting software using open-source library27. Hence, the possibility of the system has been expanded. Finally, the system provides automated high-throughput behavioral screening for long-term assays that are used to study a wide-range of cognitive functions, which is beneficial in phenotyping and validating disease models.

The development of animal models for neuronal disease over the last decade has provided a mechanistic understanding of their basis and in order to promote the therapeutic advances1,2,3. Application of a high-throughput behavioral test battery in genetic animal models is a heuristic research tool to investigate the underlying mechanisms of human diseases and identification of drug therapies. Research test batteries adapted for long-term observation of aging and/or dementia models have traditionally forced laboratories to consume great amounts of specialized manpower and time. A home-cage monitoring system would be a cost-effective strategy as it would reduce the cost of behavioral observation by humans. Some research teams have developed automated vision-based tools that assist behavioral phenotyping of a single individual in a small home cage4,5,6. However, such methods limit the social interaction, the size of testing environments, and the variety of behavioral measures that include cognitive functions. The IntelliCage is a second-generation home-cage monitoring system designed to perform various cognitive tasks in a social home cage. Importantly, this method can eliminate daily handling that enables us to perform long-term behavioral monitoring with assessment of cognitive functions, and it can eliminate the requirements for specialized practical handling, and enable highly reproducible data acquisition7. Here, we describe the long-term phenotyping and validation in genetic mouse models of Alzheimer&#39;s disease (AD) which has been generated recently8,9,10 using the automated home-cage monitoring system. A test battery, which included assessments of spatial learning and executive functions, was repeatedly performed at multiple age points (9&#8211;12 and 14&#8211;17 months old). This age-dependent phenotyping allowed us to identify the transient and/or permanent effects of genes on various cognitive aspects. We found that some AD models showed both transient and permanent phenotypes of several cognitive aspects tested in the long-term analysis using the automated home-cage monitoring system10. Thus, the automated study using home-cage monitoring system is beneficial and cost-effective for long-term behavioral phenotyping and validation in various models of cognitive dysfunction.

<table><tbody><tr><td>IntelliCage</td><td>TSE Systems</td><td>-</td><td>Parchased in 2011 or later</td></tr><tr><td>PC</td><td>Dell</td><td>Inspiron 580s</td><td>-</td></tr><tr><td>Display</td><td>Dell</td><td>SI75T-WL</td><td>-</td></tr><tr><td>ALPHA-dri</td><td>Shepherd Specialty Papers</td><td>-</td><td>Standard bedding</td></tr><tr><td>Aron Alpha (Krasy Glue) 2 g</td><td>Toagosei (Krasy Glue)</td><td>#04612</td><td>Cyanoacrylates for gluing magnet and blak arm</td></tr><tr><td>Handheld Transponder Reader</td><td>BTS-ID</td><td>R-560</td><td>Transponder reader, which reads both Trovan and DataMars</td></tr><tr><td>Transponder</td><td>DataMars</td><td>T-VA, T-VAS, or another series</td><td>Basic package of transponders and implanters</td></tr><tr><td>Diamond Grip Plus</td><td>Ansel Microflex</td><td>DGP-INT-M</td><td>Experimental glove</td></tr><tr><td>Isoflurane</td><td>Pfizer</td><td>1119701G1092</td><td>-</td></tr><tr><td>Vaporizer for small animals</td><td>DS Pharma Biomedical</td><td>SF-B01</td><td>Facemask included</td></tr><tr><td>Neo-Medrol</td><td>Pfizer</td><td>006472-001</td><td>Eye ointment</td></tr><tr><td>Ethanol (70%)</td><td>-</td><td>-</td><td>-</td></tr><tr><td>Excel</td><td>Microsoft</td><td>00202-51382-15524-AA928</td><td>For data analysis</td></tr></tbody></table>

automated, long-term behavioral assay for cognitive functions in multiple genetic models of alzheimer's disease, using intellicage

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive decline, such as Alzheimer&#39;s disease (AD), have become a promising tool to elucidate the underlying mechanisms and promote the therapeutic advances. An important step is the validation and characterization of expected behavioral abnormality in the models, in the case of AD, cognitive decline. The long-term behavioral investigations of laboratory animals to study the effect of aging demand substantial efforts from researchers. The IntelliCage system is a high-throughput and cost-effective test battery for mice that eliminates the need for daily human handling. Here, we describe how the system is utilized in the long-term phenotyping of a genetic Alzheimer&#39;s disease model, specifically focusing on the cognitive functions. The experiment employs repeated battery of tests that assess spatial learning and executive functions. This cost-effective age-dependent phenotyping allows us to identify the transient and/or permanent effects of genes on various cognitive aspects.

In our previous study, the age-dependent cognitive deficits in AD models were detected by the experiments using the automated home-cage monitoring system10. Their performance of AD models in PP was intact in both young adults and older subjects; however, the performance in PPR was significantly and progressively impaired (Figure 6). It is also important to observe the general behavior or anxiety in the adaptation phase because such traits may affect the cognition15. The AD models did not show any gross abnormalities in the visit, nosepoke, and licking numbers in the FA, NP, and DSA sessions. Thus, the AD models may have lower flexibility.
To assess the executive functions, the behavioral performance in SRT and DD was recorded. Older AD models (NL-G-F) lacked accuracy in the last stage of SRT (SRT-Test 2) (Figure 8). Facilitated compulsivity was observed in young adults and old subjects of the NL-G-F mice (Figure 9B, top). Interestingly, although there was an increase in the compulsivity of the NL-F mice who were young adults, in old age, it became comparable to that of the wild-type mice (Figure 9B, bottom). This is an example of the transient phenotype of the NL-F mutation.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/58009/58009fig1.jpg" /> 
Figure 1: Components of the automated home-cage monitoring system. (A) Overview of the system. (B) Location of connectors. (C) Corner chamber with operant door. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig1large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/58009/58009fig2.jpg" /> 
Figure 2: Example of the group housing for the study. Typically, 12 mice per cage are used. In the case of using four groups (three genetic models and one wild-type), three mice per group per IntelliCage are considered adequate. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/58009/58009fig3.jpg" /> 
Figure 3: Software for the system. (A) The &#34;Designer&#34; is used to build experimental files. Left: The Animal list part includes the information of the animal and group definitions. Right: The Module Space is used to define experimental operations. (B) The &#34;Controller&#34; can be used to run, monitor, and record experiments. Left: The status of one of the cage working with displaying animals&#39; visit, nosepoke, and licking in the four corners. Right: Alarm windows will appear if some troubles happen. (C) The Analyzer can be used to handle and export data acquired by the Controller. Left: All data are tagged with information of the animal, environment inside the cage, and time. Filtering will help further analysis in Excel or other analytic software. Right: The time-line of the visits (also nosepoke or licks) can be shown individually. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig3large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58009/58009fig4.jpg" /> 
Figure 4: Transponder implantation. (A) Microchip of the transponder (DataMars). (B) Side view of the transponder implantation (transpondering). Avoid cause injury on spinal cord. (C) Top view of the transponder. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig4large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/58009/58009fig5.jpg" /> 
Figure 5: Time-line of the experiments for the cognitive assessment with the automated home-cage monitoring system. A test battery for cognitive assessment was performed twice (1st set, 9&#8211;12 months old; and 2nd sets, 14&#8211;17 months old) followed by experiments for assessing general activity in the end (3rd set [18 months old]). This battery was designated for assessing multiple cognitive domains (indicated by colors&#160;&#8212; Red: general activity; Blue: spatial learning and memory; Green: executive function), that has advantages in validation and characterization of the expected cognitive deficits. FA: Free Adaptation; NPA: Nosepoke Adaptation; DSA: Drinking Session Adaptation; PP: Place Preference; PPR: Place Preference Reversal; SRT: Serial Reaction Time (for impulsivity and attention); PA: Place Avoidance; DD: Delay Discounting. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig5large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/58009/58009fig6.jpg" /> 
Figure 6: Experimental design and representative results of PP and PPR tasks. (A) Top, an example module design for PP or PPR. Bottom, correct corner setting is changed to opposite side in PPR. (B) Deficits in spatial reversal learning in an AD model (NL-G-F) elicited at older age. Data are expressed as mean &#177; standard error of mean (SEM). &#8727;p &#60;0.05; &#8727;&#8727;p &#60;0.01. Colors indicate groups of comparison: Blue: NL vs WT; Red: NL-F vs WT; Green: NL-G-F vs WT. This figure has been modified from reference10. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig6large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/58009/58009fig7.jpg" /> 
Figure 7: Trial flow of SRT tasks. Left: the trial flow of the SRT (imp). The first nosepoke defines the correct side, and initiates a delay period (0.5&#8211;4.0 s), after which yellow LEDs are turned on. The door is then opened. Right: the trial flow of the SRT (att). The first nosepoke defines the correct side, and initiates a delay period (2.0 s), after which yellow LEDs are turned on in shorter time (0.2&#8211;1.0 s). The mice are provided with a time period during which nosepokes are allowed (the limited hold, 2 s). The doors open (5 s) only after a correct nosepoke, which is the first nosepoke during the limited hold. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig7large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/58009/58009fig8.jpg" /> 
Figure 8: Experimental design and representative results of SRT tasks. (A) An example module design for SRT. (B) Attention decline in an AD model specific to at older age. Data are expressed as mean &#177; SEM. &#8727;p &#60;0.05; &#8727;&#8727;p &#60;0.01. Colors indicate groups of comparison: Blue: NL vs WT; Red: NL-F vs WT; Green: NL-G-F vs WT. This figure has been modified from the reference10. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig8large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/58009/58009fig9.jpg" /> 
Figure 9: Experimental design and representative results of DD task. (A) An example of the module for the DD task. (B) Facilitated compulsivity in the AD model (NL-G-F) at both young and old ages. This is an example of phenotype. On the other hand, compulsivity was transiently increased in another AD model (NL-F). Data are expressed as mean &#177; SEM. &#8727;p &#60;0.05; &#8727;&#8727;p &#60;0.01. Colors indicate groups of comparison: Blue: NL vs WT; Red: NL-F vs WT; Green: NL-G-F vs WT. This figure has been modified from the previous work10. <a href="https://www.jove.com/files/ftp_upload/58009/58009fig9large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

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Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer&#39;s disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

Multiple factors&#8212;such as aging and genes&#8212;are frequently associated with cognitive decline. Genetically modified mouse models of cognitive ...

automated-long-term-behavioral-assay-for-cognitive-functions-multiple

Nanyang Technological University

Research

JoVE Journal

Behavior

12.1K Views.  RIKEN. This paper describes a protocol for cognitive assessments for genetic models of the Alzheimer's disease using the IntelliCage system, which is a high throughput automated behavioral monitoring system with operant conditioning.

Video: Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

Automated, Quantitative Cognitive/Behavioral Screening of Mice: For Genetics, Pharmacology, Animal Cognition and Undergraduate Instruction

We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and pharmacological manipulations on basic mechanisms of cognition and learning in mice. A standard polypropylene mouse housing tub is connected through an acrylic tube to a standard commercial mouse test box. The test box has 3 hoppers, 2 of which are connected to pellet feeders. All are internally illuminable with an LED and monitored for head entries by infrared (IR) beams. Mice live in the environment, which eliminates handling during screening. They obtain their food during two or more daily feeding periods by performing in operant (instrumental) and Pavlovian (classical) protocols, for which we have written protocol-control software and quasi-real-time data analysis and graphing software. The data analysis and graphing routines are written in a MATLAB-based language created to simplify greatly the analysis of large time-stamped behavioral and physiological event records and to preserve a full data trail from raw data through all intermediate analyses to the published graphs and statistics within a single data structure. The data-analysis code harvests the data several times a day and subjects it to statistical and graphical analyses, which are automatically stored in the &#34;cloud&#34; and on in-lab computers. Thus, the progress of individual mice is visualized and quantified daily. The data-analysis code talks to the protocol-control code, permitting the automated advance from protocol to protocol of individual subjects. The behavioral protocols implemented are matching, autoshaping, timed hopper-switching, risk assessment in timed hopper-switching, impulsivity measurement, and the circadian anticipation of food availability. Open-source protocol-control and data-analysis code makes the addition of new protocols simple. Eight test environments fit in a 48 in x 24 in x 78 in&#160;cabinet; two such cabinets (16 environments) may be controlled by one computer.

Fully automated system for measuring physiologically meaningful properties of the mechanisms mediating spatial localization, temporal localization, duration, rate&#160;and probability estimation, risk assessment, impulsivity, and the accuracy and precision of memory, in order to assess the effects of genetic and pharmacological manipulations on foundational mechanisms of cognition in mice.

We describe a high-throughput, high-volume, fully automated, live-in 24/7 behavioral testing system for assessing the effects of genetic and ...

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station (INBEST)

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of chronic CNS disorders, there is an increasing demand for murine models with high validity at the behavioral level. However, multiple pathogenic mechanisms and complex functional deficits often impose challenges to reliably measure and interpret behavior of chronically sick mice. Therefore, the assessment of peripheral pathology and a behavioral profile at several time points using a battery of tests are required. Video-tracking, behavioral spectroscopy, and remote acquisition of physiological measures are emerging technologies that allow for comprehensive, accurate, and unbiased behavioral analysis in a home-base-like setting. This report describes a refined phenotyping protocol, which includes a custom-made monitoring apparatus (Integrated Behavioral Station, INBEST) that focuses on prolonged measurements of basic functional outputs, such as spontaneous activity, food/water intake and motivated behavior in a relatively stress-free environment. Technical and conceptual improvements in INBEST design may further promote reproducibility and standardization of behavioral studies.

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station INBEST

Prolonged and comprehensive monitoring of mice in a home-cage environment provides a deeper understanding of aberrant behavior in murine models of brain diseases. This paper describes the Integrated Behavioral Station (INBEST) as the key component of contemporary behavioral analysis.

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of ...

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

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

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

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

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, ...

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

Neuroscience

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Intracerebroventricular Injection of Amyloid-&#946; Peptides in Normal Mice to Acutely Induce Alzheimer-like Cognitive Deficits

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, progressive accumulation of A&#946; oligomers and plaques is observed. Such A&#946; abnormalities are believed to block long-term potentiation, impair synaptic function, and induce cognitive deficits. Clinical and experimental evidences have revealed that the acute increase of A&#946; levels in the brain allows development of Alzheimer-like phenotypes. Hence, a detailed protocol describing how to acutely generate an AD mouse model via the intracerebroventricular (ICV) injection of A&#946; is necessary in many cases. In this protocol, the steps of the experiment with an A&#946;-injected mouse are included, from the preparation of peptides to the testing of behavioral abnormalities. The process of preparing the tools and animal subjects before the injection, of injecting the A&#946; into the mouse brain via ICV injection, and of assessing the degree of cognitive impairment are easily explained throughout the protocol, with an emphasis on tips for effective ICV injection of A&#946;. By mimicking certain aspects of AD with a designated injection of A&#946;, researchers can bypass the aging process and focus on the downstream pathology of A&#946; abnormalities.

The amyloid-&#946; (A&#946;)-injected animal model enables the administration of a defined quantity and species of A&#946; fragments and reduces individual differences within each study group. This protocol describes the intracerebroventricular (ICV) injection of A&#946; without stereotactic instruments, enabling the production of Alzheimer-like behavioral abnormalities in normal mice.

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, ...

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 ...

Medicine

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. The paradigm includes tests for spontaneous alternation in a Y-maze, novel object recognition, and limb clasping. These tests were selected because they: 1) interrogate function of cognitive or motor domains and the correlate neural circuitry relevant to the human disease state, 2) have clearly defined endpoints, 3) have easily implementable quality control checks, 4) can be run in a moderate throughput format, and 5) require little intervention by the investigator. These methods are designed for investigators looking to screen compounds for activity in short-term and working memory tasks, or functional motor behaviors associated with Alzheimer's disease mouse models. The methods described here use behavioral tests that engage a number of different brain regions including hippocampus and various cortical areas. Investigators that desire cognitive tests that specifically assess cognition mediated by a single brain region could use these techniques to supplement other behavioral tests.

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional ...

Shuttle Box Assay as an Associative Learning Tool for Cognitive Assessment in Learning and Memory Studies using Adult Zebrafish

Cognitive deficits, including impaired learning and memory, are a primary symptom of various developmental and age-related neurodegenerative diseases and traumatic brain injury (TBI). Zebrafish are an important neuroscience model due to their transparency during development and robust regenerative capabilities following neurotrauma. While various cognitive tests exist in zebrafish, most of the cognitive assessments that are rapid examine non-associative learning. At the same time, associative-learning assays often require multiple days or weeks. Here, we describe a rapid associative-learning test that utilizes an adverse stimulus (electric shock) and requires minimal preparation time. The shuttle box assay, presented here, is simple, ideal for novice investigators, and requires minimal equipment. We demonstrate that, following TBI, this shuttle box test reproducibly assesses cognitive deficit and recovery from young to old zebrafish. Additionally, the assay is adaptable to examine either immediate or delayed memory. We demonstrate that both a single TBI and repeated TBI events negatively affect learning and immediate memory but not delayed memory. We, therefore, conclude that the shuttle box assay reproducibly tracks the progression and recovery of cognitive impairment.

Learning and memory are potent metrics in studying either developmental, disease-dependent, or environmentally induced cognitive impairments. Most cognitive assessments require specialized equipment and extensive time commitments. However, the shuttle box assay is an associative learning tool that utilizes a conventional gel box for rapid and reliable assessment of adult zebrafish cognition.

Cognitive deficits, including impaired learning and memory, are a primary symptom of various developmental and age-related neurodegenerative diseases and ...

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage

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