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Behavioral Characterization of an Angelman Syndrome Mouse Model

Published: October 20th, 2023



1Czech Centre for Phenogenomics, Institute of Molecular Genetics of the Czech Academy of Sciences, 2Laboratory of Transgenic Models of Diseases, Institute of Molecular Genetics of the Czech Academy of Sciences

This manuscript presents a set of highly reproducible behavioral tests to validate an Angelman syndrome mouse model.

This manuscript describes a battery of behavioral tests available to characterize Angelman syndrome (AS)-like phenotypes in an established murine model of AS. We use the rotarod learning paradigm, detailed gait analysis, and nest building test to detect and characterize animal motor impairments. We test animal emotionality in the open field and elevated plus maze tests, as well as the affect in the tail suspension test. When AS mice are tested in the open field test, the results should be interpreted with care, since motor dysfunctions influence mouse behavior in the maze and alter activity scores.

The reproducibility and effectiveness of the presented behavioral tests has already been validated in several independent Uba3a mouse lines with different knockout variants, establishing this set of tests as an excellent validation tool in AS research. Models with the relevant construct and face validity will warrant further investigations to elucidate the pathophysiology of the disease and grant the development of causal treatments.

Angelman syndrome (AS) is a rare neurodevelopmental disease. The most common genetic origin of AS is a large deletion of the 15q11-q13 region of the maternally-derived chromosome, which is found in nearly 74% of patients1. Deletion of this region causes the loss of UBE3A, the main causative gene of AS that encodes an E3 ubiquitin ligase. The paternal allele of the UBE3A gene in neurons is silenced in a process known as imprinting. As a consequence, paternal imprinting of the gene allows only maternal expression in the central nervous system (CNS)2. Therefore, UBE3A gene deletion from the matern....

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All animals and experiments used in this study underwent ethical review and were conducted in accordance with the European Directive 2010/63/EU. The study was approved by the Czech Central Commission for Animal Welfare. Mice were housed in individually ventilated cages and maintained at a constant temperature of 22 ± 2 °C with a 12 h light/dark cycle. The mice were provided with mouse chow and water ad libitum. The mice were housed in groups of three to six animals per cage. No handling other than weig.......

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Elevated plus maze and open field tests
The EPM and OF tests use the natural tendency of rodents to explore new environments18,19. The exploration is governed by an approach-avoidance conflict, where rodents choose between the exploration of a new environment and avoidance of possible danger. Animals explore unknown places in search for shelter, social contact, or foraging. However, new places may involve risk factors such as predators or c.......

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AS models created in different murine strains are commonly validated with tests of animal emotional state, motor functions, and cognitive abilities to facilitate comparison to human symptoms31,32. A motor deficit in AS models is the most consistent finding across laboratories, followed by an unchanged emotionality state of mutants and difficulties building nests31,32,33........

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This research was supported by the Czech Academy of Sciences RVO 68378050, LM2018126 Czech Centre for Phenogenomics provided by MEYS CR, OP RDE CZ.02.1.01/0.0/0.0/16_013/0001789 (Upgrade of the Czech Centre for Phenogenomics: developing toward translation research by MEYS and ESIF), OP RDE CZ.02.1.01/0.0/0.0/18_046/0015861 (CCP Infrastructure Upgrade II by MEYS and ESIF), and OP RDI CZ.1.05/2.1.00/19.0395 (higher quality and capacity for transgenic models by MEYS and ERDF). In addition, this study received funding from the NGO "Association of Gene Therapy (ASGENT)", Czechia ( and LM2023036 Czech Centre for Phenogenomics provided by Ministry....

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NameCompanyCatalog NumberComments
Cages, individually ventilatedTechniplast
DigiGaitMouse Specifics, Inc., 2 Central Street Level
Unit 110
Framingham, MA 01701, USA
Equipment was tendered, no catalogue  number was provided, nor could be find on company's web siteDetailed analysis of mouse gait, hardware and software provided. 
FDA Nestlet squaresDatesand Ltd., 7 Horsfield Way, Bredbury, Stockport SK6, UKMaterial was bought from Velaz vendor via direct email request. Velaz do not provide any catalogue no.Cotton nestlets for nest building test. Nestlet discription: 2-3 g each, with diameter around 5 x 5 x 0.5cm.
Mouse chowAltramion
RotarodTSE Systems GmbH, Barbara-McClintock-Str.4
12489 Berlin, Germany
Equipment was tendered, no catalogue  number was provided, nor could be find on company's web siteRotarod for 5 mice, hardware and software provided. Drum dimensions: Diameter: 30 mm, width per lane: 50 mm, falling distance 147 mm.
Tail Suspension TestBioseb, In Vivo Research Instruments, 13845 Vitrolles
Reference: BIO-TST5Fully automated equipment for immobility time evaluation of 3 mice hanged by tail, hardware and software provided
Transpore medical tapeMedical M, Ltd.P-AIRO1291The tape used to attach an animal to the hook by its tail.
Viewer - Video Tracking SystemBiobserve GmbH, Wilhelmstr. 23 A
53111 Bonn, Germany
Equipment with software were tendered, no catalogue  number was provided, nor could be find on company's web siteSoftware with custom made hardware: maze, IR base, IR sensitive cameras. Custom-made OF dimensions: 42 x 42 cm area, 49 cm high wall, central zone area: 39 cm2. A custom-made EPM was elevated 50 cm above the floor, with an open arm 79 cm long,  9 cm wide, and closed arm 77 cm long, 7.6 cm wide. 

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