전임상 웹사이트임상 웹사이트

타우 병증의 AAV-Tau 마우스 모델

인간 MAPT AAV 벡터를 주입하여 타우 응집체, 신경염증, 신경퇴행 및 운동장애를 동반한 도파민성 탈신경을 생성합니다.

AAV 타우병증(AAV-Tau) 모델 개요

이 진행성 핵상마비 및 피질-기저변성 모델의 경우, 생쥐의 약 2-3개월령에 C57BL/6 마우스의 흑질에 야생형 인간 타우(MAPT)를 과발현하는 AAV를 일측성 정위 주사합니다. 이 마우스 모델은 다음과 같은 인간 타우병의 몇 가지 주요 특징을 재현합니다.

  • 흑질 소구체의 도파민성 뉴런 감소
  • 동측 선조체의 도파민 신경절 절제
  • 세포체와 신경돌기에 있는 인산화 타우의 응집체
  • 활성화된 미세아교세포
  • 반응성 성상세포
  • 운동 기능 장애
  • 생체 내 MRI 스캔으로 측정한 뇌 위축(흑질, 중뇌, 후두핵)

AAV-Tau 모델 생성

모델 생성을 위한 일반적인 스키마는 다음과 같습니다.

설치류에 대한 정위 수술 준비

이 특정 모델의 경우, 생후 8-12주령의 C57BL/6 마우스를 사용합니다. 그런 다음, AAV 벡터를 흑질 부근에 정위 주사합니다. 높은 정확성과 정밀성을 위해 자동 마이크로 주사기가 장착된 디지털 정위 장치를 사용합니다.

이 모델을 이용한 연구는 신속하게 시작할 수 있습니다. 생체 내 단계는 일반적으로 약 6주 동안 지속됩니다. 따라서, 특히 기존의 알츠하이머병 및 타우병에 대한 타우 트랜스제닉 모델과 비교할 때, 비교적 짧은 시간 내에 결과를 얻을 수 있습니다.

검증된 조치

  • 뒷다리 쥐기 테스트
  • 테일 서스펜션 스윙 테스트
  • 실린더 테스트
  • 로타로드 테스트
  • MRI 뇌 위축
  • IHC & 멀티플렉스 면역 형광법

 

주입된 SNc의 AT8 면역 형광 염색 
주입된 SNc의 Iba-1 및 GFAP 이중 면역 형광 염색
동측 반구에서 티로신 하이드 록 실라 제 염색의 극적인 감소
주입된 SNc에서 도파민 신경세포의 심각한 손실
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주입된 SNc의 AT8 면역 형광 염색 

주입된 SNc의 Iba-1 및 GFAP 이중 면역 형광 염색

동측 반구에서 티로신 하이드 록 실라 제 염색의 극적인 감소

주입된 SNc에서 도파민 신경세포의 심각한 손실

모델 특성화

아래의 대화형 프레젠테이션을 통해 생체 데이터와 전체 다중 면역형광 조직 섹션의 고해상도 이미지를 포함하여 AAV-Tau 마우스 모델의 특징을 살펴볼 있습니다.

왼쪽 패널을 사용하여 이 "이미지 스토리" 간단하게 탐색할 수 있습니다.

왼쪽 마우스 버튼을 사용하여 고해상도 현미경 이미지를 이동할 있습니다. 마우스/트랙패드(위/아래) 또는 왼쪽 상단 모서리에 있는 + 및 - 버튼 사용하여 확대 축소할 있습니다 . 오른쪽 상단 모서리에 있는 제어판에서 채널 및 세분화에 대한 이미지 설정을 토글(켜기/끄기), 색상 변경 및 조정할 수 있습니다.

최상의 상호작용 경험을 위해 전체 화면 모드를 사용하는 것이 좋습니다.

Characterization of a Novel AAV-hTau Mouse Model of Tauopathies with Parkinsonian Features

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Biospective Preclinical Logo

Tauopathies, such as Progressive Supranuclear Palsy and Corticobasal Degeneration, are rare diseases with prominent Parkinsonian features, including motor symptoms such as postural instability, vertical gaze palsy, rigidity, slowed movement (bradykinesia), muscle contractions (dystonia), and sudden jerks (myoclonus). Furthermore, individuals may face difficulties with speech and swallowing, cognitive decline, and loss of sensory perception at the cortical level. These neurodegenerative diseases are often rapidly progressing and pathologically characterized by phosphorylated tau inclusions in neurons and glia.

A significant problem for the development of disease-modifying therapeutics for tauopathies is the lack of animal models that recapitulate the human disease. To address this issue, Biospective has developed and characterized an adeno-associated virus (AAV) vector-induced mouse model that is well-suited for preclinical therapeutic efficacy studies for Progressive Supranuclear Palsy and Corticobasal Degeneration.

This Interactive Presentation illustrates some of the key motor function, neuroimaging, and pathologic features of Biospective's AAV human tau model of tauopathies with Parkinsonian features.

This model was generated by injecting 2 month-old C57BL/6 mice with AAV-hTau (wild-type 2N4R human tau) or AAV-null (control) vectors unilaterally into the left substantia nigra pars compacta (SNc) using a digital stereotaxic device with an automated microinjector.

Coronal Atlas with AAV Injection Site

Coronal Atlas View of SNc Injection Site

Multiplex immunofluorescence (mIF) images were generated by immunostaining for phosphorylated Tau (AT8), GFAP, Iba-1, Tyrosine Hydroxylase, Dopaminergic Nuclei, and counterstained with the DAPI nuclear stain. Tissue sections were digitized using a high-throughput slide scanner and were processed using Biospective's PERMITSTM software platform.

To navigate though this Image Story, you can use the arrows and/or the Table of Contents icon in the upper right corner of this panel.

Navigation Panel with Tooltips

You can also interact with the microscopy image in the viewer on the right at any time to further explore this high-resolution data.

Phosphorylated Human Tau Pathology

This microscopy image shows AT8 immunostaining for pTau. The ipsilateral (left hemisphere) midbrain shows extensive staining in the vicinity of the SNc and slightly beyond. For anatomical reference, an illustration with atlas labels for this brain level is provided below.

Coronal Brain Atlas at the Level of the Substantia Nigra

Coronal Mouse Brain Section (Bregma -3.2) with Neuroanatomy Labels

Tau Pathology in Neuronal Cell Bodies & Processes

This high magnification image shows extensive pTau staining in both the soma and processes of neurons in the SNc.

Neurodegeneration in the Substantia Nigra

As can be seen in this microscopy image, there is substantial loss of TH-positive dopaminergic neurons in the ipsilateral SNc compared to the contralateral hemisphere. For reference, an illustration with atlas labels for this brain level is provided below.

Coronal Mouse Brain Section (Bregma -3.2) with Neuroanatomy Labels

Using our PERMITSTM quantitative analysis software, we have quantified the TH staining in the SNc. The plot below shows a highly significant reduction in the ipsilateral hemisphere of the AAV-Tau compared to the AAV-null (control) mice.

Tyrosine Hydroxylase Staining in the Substantia Nigra

TH stain density for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, **** p<0.0001.

Brain Atrophy in the SNc and Midbrain

Regional brain atrophy is a key feature of tauopathies. Magnetic Resonance Imaging (MRI) is clinically used for non-invasive neuroimaging of Progressive Supranuclear Palsy (see our Resource) and Corticobasal Degeneration (see our Resource). Our team at Biospective has investigated the spatiotemporal pattern of brain atrophy in tauopathies (see MRI Measures of Disease Progression for Progressive Supranuclear Palsy Clinical Trials and MRI & Corticobasal Degeneration). We have found significant atrophy in multiple brain areas, including the midbrain and striatum in both diseases.

Given that MRI is a “translational biomarker”, we have acquired high-resolution in vivo anatomical 3D MR images from the AAV-hTau and AAV-null (control) mice using a 7T preclinical MRI scanner. We performed fully-automated image processing using our proprietary NIGHTWINGTM software and found highly significant brain atrophy in the SNc and midbrain. This data corresponds nicely to the loss of TH-positive neurons seen in the microscopy image.

MRI Brain Atlas and Volume Data for the SNc Level

Anatomical MRI with segmented SNc and midbrain, as well as plots of relative difference between ipsilateral and contralateral hemispheres for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, **** p<0.0001.

Dopaminergic Neurons in the Contralateral SNc

This microscopy image shows the contralateral (right hemisphere) SNc which demonstrates TH-positive cell bodies and processes in red. The nuclei of the dopaminergic neurons are shown in blue.

Loss of Dopaminergic Neurons in the Ipsilateral SNc

This microscopy image shows the ipsilateral (left hemisphere) SNc which demonstrates a profound reduction of TH-positive cell bodies and processes (in red) compared to the contralateral hemisphere. The dopaminergic neuron nuclei are shown in blue.

Neurodegeneration in the Caudate-Putamen & Dopaminergic Motor Deficits

This microscopy image shows severe dopaminergic denervation of the ipsilateral (left hemisphere) caudate-putamen (loss of TH-positive terminals). For reference, an illustration with atlas labels for this approximate brain level is provided below.

Atlas Section at the Striatum Level

Coronal Mouse Brain Section (Bregma +0.86) with Neuroanatomy Labels

Using our PERMITSTM quantitative analysis software, we have quantified the TH staining in the Caudate-Putamen. The plot below shows a highly significant reduction in the ipsilateral hemisphere.

Tyrosine Hydroxylase Staining in the Caudate-Putamen

TH stain density for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, **** p<0.0001.

This loss of dopaminergic innervation corresponds well with unilateral motor deficits in these mice, including a highly significant increase in use of the ipsilateral paw during the Cylinder Test, decreased latency to fall in the Rotarod Test, increased swings to the contralateral side in the Tail Suspension Swing Test (TSST), and increased Hindlimb Clasping.

Illustration of Motor Tests and Plots of AAV-Tau vs. AAV-null

Cylinder Test, Rotarod Test, Tail Swing Suspension Test (TSST), and Hindlimb Clasping data for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, **** p<0.0001.

Loss of Dopaminergic Terminals in the Ipsilateral Caudate-Putamen

This high magnification view shows the severe extent of loss of dopaminergic (TH-positive) terminals in the ipsilateral striatum. There are some remaining (albeit dystrophic) axons present.

We have also identified significant brain atrophy in the caudate-putamen on MRI scans, which aligns well with our analysis of human MRI data from Progressive Supranuclear Palsy and Corticobasal Degeneration populations. This data supports the “translatability” of this tauopathy model.

MRI Atlas and Volume Data at Striatum Level

Anatomical MRI with segmented striatum, as well as plot of relative difference between ipsilateral and contralateral striatum. ****p<0.0001.

Microgliosis in Response to Human 2N4R Tau Expression

In this low magnification image, one can readily appreciate the higher density of Iba-1-positive microglia in the ipsilateral (left) hemisphere (indicated by the box) relative to the contralateral hemisphere.

The plot below shows the Iba-1 stain density in the SNc.

Plots of Iba-1 staining for AAV-Null and AAV-Tau Injected Mice

Iba-1 stain density for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, *** p<0.001.

We have performed a morphological analysis of microglia using a novel computer vision & machine learning approach developed by our team. This fully-automated algorithm classifies non-activated (ramified) and activated (non-ramified) microglia.

Image of non-activated and activated microglia

The plot below shows the microglial activation in the SNc, with highly significant increased microglial activation in the AAV-Tau mice.

Plot of PERMITS Data Showing Activated Microglia in SNc

Microglial activation for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, *** p<0.0001.

Iba-1 Staining in Proximity to Phosphorylated Tau

This high magnification view shows the increased density of Iba-1-stained microglia in areas with phosphorylated tau aggregates.

Astrogliosis & Human Tau Pathology

This low magnification microscopy image show a higher density of GFAP-positive astrocytes in the ipsilateral hemisphere (indicated by the box). The plot below shows the GFAP stain density in the SNc.

Plot of GFAP Staining Density in SNc

GFAP stain density for AAV-Tau compared to AAV-null (control) injections; mean ± SEM, t-test, **** p<0.0001.

GFAP Staining in Proximity to p-Tau

This high magnification view shows the increased density of GFAP-stained astrocytes in areas with phosphorylated Tau aggregates.

Summary

This novel mouse model of tauopathies with Parkinsonian features recapitulates many of the hallmark features of Progressive Supranuclear Palsy and Corticobasal Degeneration, including the development of asymmetric motor dysfunction (due to unilateral injection), and associated loss of TH+ SNc neurons and striatal TH expression.

AAV-hTau regionally results in highly significant brain atrophy, elevated microglial density and activation levels, and increased astrocyte density and hypertrophy. Further studies are planned to continue to investigate the pathologic changes in this model.

This inducible and rapidly progressing mouse model is well-suited for drug discovery with quantitative in vivo and ex vivo readouts, and possesses distinct advantages over existing transgenic models as a screening method for novel treatment options targeting tau-related pathology.

Please feel free to further explore the microscopy image in the viewer.

We would be happy to discuss this model and our characterization if you would like to Contact Us.

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AAV-Tau 마우스 모델의 특성, 검증된 측정 방법, 전임상 신경과학 CRO 서비스에 대해 자세히 알아보세요.

더 많은 알츠하이머병 및 타우병증 모델 알아보기

자주 묻는 질문

MRI 뇌 위축은 번역적 바이오마커로 사용될 수 있습니까?


뇌 위축은 주로 타우 단백질 또는 아밀로이드 베타에 의해 유발됩니까?


이 타우병 모델에서 도파민성 신경절의 도파민 분비를 정량적으로 평가할 수 있습니까?


"꼬리 매달기 테스트"는 무엇입니까?


AAV-Tau 모델에서 "반응성" 성상교세포를 어떻게 정량화합니까?


"아데노바이러스 관련 바이러스"(AAV) 벡터란 무엇입니까?


관련 콘텐츠

알츠하이머병 및 타우병에 대한 최신 정보와 동물 모델에서 치료제 평가를 위한 번역적 바이오마커 사용과 관련된 모범 사례.

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