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).

All the procedures were approved by the institutional animal care and use committee, and they were carried out according to the RIKEN Brain Science Institute&#8217;s guidelines for animal experimentation.
1. Setting Apparatus
NOTE: An overview of the automated home-cage monitoring system is shown in Figure 1. Each system (39 cm x 58 cm x 21 cm) contains one microprocessor, and four corner chambers, each of which has two water bottles and ...

<ol>
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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 (...

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) 2g</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 tra...

Watch this Scientific Journal Video about Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer's Disease, Using IntelliCage at JoVE.com

Automated, Long-term Behavioral Assay for Cognitive Functions in Multiple Genetic Models of Alzheimer&#039;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.

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 ...

This method can help answer key questions in the neuroscience field, such as animal models of cognitive deficits, or psychiatric diseases. The main advantage of this technique is that highly reproducible data of variety of cognitive tasks can be obtained with reduced manpower. Generally, individuals new to this method will struggle because the system needs many steps for the accurate work.The designer software is where the experimental file which contains key experimental definitions, is produced. The experimental files are used for running specific tests, such as price preference, where only one of the four corners is not accessible within a specific time window. Begin by creating the animal list.Define the conditions and set the parameters for the number of subject mice, number of genetic lines, sex of the animals, and number of the cage to be used. Then, select the appropriate transponder type, DataMars or Trovan in the central toolbar. Next, in the Groups panel, add or remove the experimental groups by pressing the green plus or red cross button in the Groups window respectively.Use the Cluster function to operate subgroups equally by defining correct, incorrect, and neutral corners and sides. Then site variables including Name, Tag, Sex, Group, and Cluster. Save and paste the animals list by selecting Export Animals.And Import Animals in the File menu bar to replicate the animal lists for another experiment. Next, set up all systems using their corresponding ID numbers in the Setup tab. Correspond the number of addresses in the designer section to the actual number of addresses.Then, build the experimental protocols in the Intellicage tab, using the following Module and Option tabs. Design the Experimental Structures in the module space by clicking the Module tab. To add new modules, press the Add, drag the units shown in the units part into the module space.To make a Nosepoke Adaption Model, drag the Door unit from the Task section, Gate, and Timer units from the Utils section, and the Visit and Nosepoke units from the Events section into the module space. Link Any on the online of the Nosepoke unit to In in the Gate unit. Link Out to Close in the Gate unit.Link Out in the Gate to Open in the Door unit. Link Out in the Gate to Activate in the Timer unit. Link Out in the Timer unit to Close in the Door unit.Set Period as 5, 000 milliseconds, in the Timer section. Define the initial status of the doors and cages in the Options Tab. Specify all doors to be closed in the non-drinking session as the typical initial status for PP or PPR tasks.Then, set the time schedules in the Options Tab with correspondence to the link in the Modules section. After defining the parameters, save the experimental file with clicking the Save As button. Lastly, load the experimental file, press the Start button of the controller, and monitor and visualize the current status of the system and the mice.To implant the radio frequency identification transponders, begin by pinching and lifting the skin around the posterior part of the scapula to create a pocket. Next, douse the injection site with 70%ethanol to minimize the introduction of hair into the subcutaneous space. Then, insert the injecting needle into the skin parallel to the spine.Eject the microchip subcutaneously and pinch the microchip through the skin to keep it in the interscapular space. Withdraw the needle slowly, and continue to pinch the area for a few seconds to provide homeostasis. Finally, return the mouse to the home cage once it has become fully ambulatory.Subjects used here were multiple AD Models, including NLF, Mild Model, and NLGF Severer Model, and two control groups. Results indicated that performance of AD Models in Place preference reversal, was mildly impaired in young adults. However, the performance in older subjects was significantly and progressively impaired.In regards to executive function, the Severer AD Model lacked accuracy in the last stages of the serial reaction time test. Facilitated compulsivity was observed in young adults and old subjects of the NLGF mice. Interestingly, although there was an increase in the compulsivity of the NLF mice who were young adults, in old age, it became comparable to that of the wild type mice.While attempting this procedure, it's important to remember to continue checking the system daily, paying special attention to the condition of animals endure inside the cages. During this procedure of the metal like drug treatment can be performed in order to answer additional questions like asking the therapeutic strategies. After its development, this technique paved the way for researchers in the fields of neuroscience to explore behavioral phenotyping, or variational in biosynthetic models.

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11.9K visualizações.  RIKEN. This method can help answer key questions in the neuroscience field, such as animal models of cognitive deficits, or psychiatric diseases. The main advantage of this technique is that highly reproducible data of variety of cognitive tasks can be obtained with reduced manpower. Generally, individuals new to this method will struggle because the system needs many steps for the accurate work.The designer software is where the experimental file which contains key experimental definitions, ...