elevated plus maze test in an angelman syndrome mouse model

행동 테스트를 사용하여 Angelman Syndrome의 Murine 모델 검증

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 maternally-derived chromosome leads to the development of AS symptoms. In humans, AS manifests at around 6 months of age, with developmental retardation that persists throughout all developmental stages and results in severe debilitating symptoms in affected individuals3,4. The core symptoms of the disorder include the deficit of fine and gross motor skills, including jerky ataxic gait, serious speech impairment, and intellectual disability. Approximately 80% of AS patients also suffer from sleep disturbances and epilepsy. To date, the only available treatment are symptomatic drugs, which reduce epileptic seizures and improve sleep quality1. Therefore, the development of robust animal models with reproducible behavioral phenotypes alongside refined phenotyping analysis will be essential to elucidate the pathophysiological mechanisms of the disorder and discover effective medications and treatments.
The complexity of the human disorder affecting the CNS demands model organisms to possess a comparable genome, physiology, and behavior. Mice are popular as a model organism due to their short reproductive cycle, small size, and relative ease of DNA modification. In 1984, Paul Willner proposed three basic disease model validation criteria: the construct, face, and predictive validity, which are used to determine the model&#39;s value5. Simply, construct validity reflects the biological mechanisms responsible for the disorder development, face validity recapitulates its symptoms, and predictive validity describes the model response to therapeutic drugs.
To adhere to the above principles, we have chosen the most common genetic etiology, a large deletion of the maternal 15q11.2-13q locus including the UBE3A gene, to create AS model mice. We used the CRISPR/Cas9 technique to delete a 76,225 bp long region spanning the entire UBE3A gene, encompassing both the coding and non-coding elements of the gene, in mice from a C57BL/6N background6. We then bred the animals to obtain UBE3A+/&#8722; heterozygous mice. For face validation of the model, we used animals from crosses of UBE3A+/&#8722; females and wild-type males to gain UBE3A+/- progeny (strain named C57BL/6NCrl-UBE3A&#60;em1(IMPC)Ccpcz&#62;/Ph and later assigned as UBE3AmGenedel/+) and control littermates. We tested their fine and gross motor skills, emotionality, and affect to recapitulate core AS symptoms. In a previous article, we also evaluated the animals&#39; cognitive functions, as AS patients also suffer from intellectual disability6. However, we found no cognitive impairments in UBE3AmGenedel/+ mice, perhaps due to the young age of the animals at the time of testing7. Later examination of the older animals, around 18 weeks old, revealed a deficit in behavioral flexibility during reversal learning in the place preference paradigm. However, the complexity of the employed equipment for this analysis requires a separate methodological module and it is not included here.
The behavioral tests presented here belong to the common phenotyping tools in genetic research, thanks to their high predictive value and sufficient construct validity8,9,10. We used these tests to validate a mouse model of AS by recapitulating core symptoms of the human disease in a reproducible, age-independent manner. The emotionality of the animal was evaluated in the elevated plus maze and open field tests. Both of these tests are based on the approach-avoidance conflict, where animals explore a new environment in search of food, shelter, or mating opportunities while simultaneously avoiding anxiogenic compartments11. Additionally, the open field test is used to test a mouse&#39;s locomotor activity8. The tail suspension test is widely used in depression research to screen for new antidepressant drugs or depressive-like phenotypes in mouse knockout models12. This test evaluates the despair that animals develop over time in an inescapable situation. Motor learning and detailed gait characteristics were determined on the rotarod and in DigiGait, respectively. Animal endurance on the accelerating rod characterizes its balance and movement coordination skills, while detailed analysis of a mouse&#39;s step patterns is a sensitive evaluation of neuromuscular impairments connected to many neurogenerative movement disorders13,14,15. The nestlet shredding test is part of the standard methodology for detecting impulsive behavior in rodents, and since it utilizes natural rodent building behavior, it indicates the animal&#39;s well-being16,17.
The size of the experimental groups was a result of a compromise to meet the 3R rule demands and efficient usage of colony breeding performance. However, to obtain statistical power, the groups had no fewer than 10 individuals, due to the establishment of a sufficient amount of breeding pairs. Unfortunately, breeding performance did not always result in a sufficient number of animals.

저자 스포트라이트: Angelman Syndrome 쥐 모델을 검증하기 위한 고도로 재현 가능한 행동 테스트 배터리 

Angelman 증후군 마우스 모델의 행동 특성

Watch this Scientific Journal Video about Elevated Plus Maze Test in an Angelman Syndrome Mouse Model at JoVE.com

Elevated Plus Maze Test in an Angelman Syndrome Mouse Model  

Angelman 증후군 마우스 모델의 Elevated Plus Maze Test

시작하려면 카메라 바로 아래 테스트 플랫폼에 플러스 모양의 미로를 놓습니다. 벽에 있는 전위차계를 사용하여 조정하는 동안 센서가 미로 중앙에 배치된 룩도미터를 사용하여 중앙에서 광도를 70럭스로 조정합니다. 뷰어 소프트웨어 아이콘을 두 번 클릭하여 소프트웨어를 열고 구성 탭의 왼쪽 상단에 있는 아이콘을 클릭하여 Elevated Plus Maze 또는 EPM 테스트를 위한 구성을 로드합니다.실험 탭의 해당 필드에 동물 ID, 유전자형, 성별, 실험 정보 등의 동물 정보를 입력합니다. 영역의 위치, 열린 팔 및 닫힌 팔이 올바르게 구성되었는지 확인합니다. 시각적 컨트롤 및 컴퓨터 마우스를 사용하여 가상 외곽선 영역이 해당 EPM 영역과 일치하는지 확인합니다.획득 탭의 왼쪽 상단에 있는 화살표 아이콘에 마우스 커서를 놓습니다. 홈 케이지에서 동물을 제거하고 EPM 중앙에 부드럽게 놓습니다. 컴퓨터 마우스를 마우스 왼쪽 버튼으로 클릭하여 실험을 시작합니다.5분 후 확인을 클릭하여 기록된 데이터를 저장합니다. 파일 이름을 지정하고 저장을 클릭합니다. 데이터 분석 탭의 왼쪽 세로 패널에 있는 아이콘을 클릭하여 오프라인 분석을 위해 결과를 csv 파일로 내보냅니다.EPM 테스트에서 불안 유사 행동과 관련된 매개변수 중 어느 것도 모계 이형접합체 Ube3A 돌연변이에서 변경되지 않았습니다.

elevated-plus-maze-test-in-an-angelman-syndrome-mouse-model

Research

JoVE Journal

Behavior

Elevated Plus Maze Test in an Angelman Syndrome Mouse Model

449 조회수. 시작하려면 카메라 바로 아래 테스트 플랫폼에 플러스 모양의 미로를 놓습니다. 벽에 있는 전위차계를 사용하여 조정하는 동안 센서가 미로 중앙에 배치된 룩도미터를 사용하여 중앙에서 광도를 70럭스로 조정합니다. 뷰어 소프트웨어 아이콘을 두 번 클릭하여 소프트웨어를 열고 구성 탭의 왼쪽 상단에 있는 아이콘을 클릭하여 Elevated Plus Maze 또는 EPM 테스트를 위한 구성을 ?...

비디오: Elevated Plus Maze Test in an Angelman Syndrome Mouse Model  

Behavioral Characterization of 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.

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 ...

연구

행동분석학

Open Field Test in an Angelman Syndrome Mouse Model

Angelman 증후군 마우스 모델의 오픈 필드 테스트

Tail Suspension Test in an Angelman Syndrome Mouse Model

Angelman 증후군 마우스 모델의 꼬리 서스펜션 테스트

Gait Analysis in an Angelman Syndrome Mouse Model

Angelman 증후군 마우스 모델의 보행 분석

Rotarod Test in an Angelman Syndrome Mouse Model

Angelman 증후군 마우스 모델의 Rotarod 테스트

Nestlet Shredding: Nest Building Test in an Angelman Syndrome Mouse Model

네슬렛 파쇄 : Angelman 증후군 마우스 모델의 둥지 건물 테스트