이 작품은 괴테 대학 병원 프랑 크푸르트와 UC에 대한 독일 연구 재단의 교부금에 의해 지원됩니다

<ol>
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저자가 공개하는 게 없다.

이전 연구에서 실험 형 간염은 주로자가 면역 질환 간 질환에 대해서만 일시적 많은 현재의 모델은 다소 복잡한 질병 유도 프로토콜 (서평 3,5 참조)에 따라 달라집니다 것으로 보도되었다. 예를 들어, 몇몇 모델은 특정 대상 항원을 표현 형질 전환 생쥐를 사용하고 adoptively 표적 항원에 특정한, 대부분 TcR - 유전자 변형, T 세포 14,15을 양도. 종종 livertropic 바이러스, 박테리아 또는 기생충과 추가 감염 질병 16-18를 유도해야합니다. 또는 CYP2D6 19를 포함하여 대상 항원, 인코딩을 plasmids과 DNA-접종이 간염을 유발하는 데 사용됩니다. 그러나 이러한 IL-12과 같은 프로 염증성 크린 시토킨, 인코딩을 plasmids있는 추가 접종은 20 필요합니다. 불행하게도, 몇 가지 예외 15-20 형 간염은 일시적입니다. CYP2D6 모델 6,21는 직접적인 방법 t를 사용단순히 hCYP2D6, AIH-2 7,8의 주요 autoantigen을 표현 아데노 바이러스와 쥐를 감염에 의해 면역 간염을 유발 O. 아데노 바이러스 구조에 의한 CYP2D6의 배달 간의 직접 타겟팅을뿐만 아니라 관용의 분해를 촉진 지역 염증을 모두 보장합니다. 그것은 바이러스 titer뿐만 아니라 행정의 노선이 모두이 모델의 중요한는 것을주의하는 것이 중요합니다. 한편, 광고-2D6은, 따라서, 복제 결함 바이러스 바이러스의 충분한 높은 titer는 간장 내에서 항원의 중요한 금액을 생성하기 위해 필요합니다. 한편, 너무 높은 titer는 치명적인 급성 간경화가 발생할 수 있습니다. 또한, 그것은 아데노 바이러스의 높은 titers에 의한 생쥐의 감염이 면역 반응 22 기능적인 피로로 연결되는 이전에 입증되었습니다. 우리의 모델에서, 우리는 둘 다 만성 세포 침투뿐만 아니라 내선 번호를 얻기 위해서는 정맥 주사와 intraperitoneal 감염의 조합을 사용ensive 섬유증. 우리는 혼자 정맥 주사 감염이 여전히 심한 세포 침윤을 초래하지만, intraperitoneal 주사로 인해 해당 복막 염증을 나타내는 subcapsular 지역에 대한 섬유증은, 콜라겐의 지속적인 subcapsular 축적 (Hintermann &amp; 카누, 준비 원고)에 관여하지 있기 관찰했습니다. 그러나 우리가 광고-GFP 또는 광고 제어 23 평등 바이러스 titers으로 감염 후 간장 별 모양의 세포와 콜라겐 증착의 활성화를 검색하지 않았으므로 관찰된 간장 섬유증은 항원에 특정한된다는 점에 유의하는 것이 중요합니다. CYP2D6 모델과 같은 영구적인 세포 침윤과 간 섬유증 6 특징으로 만성 간염과 같은 인간의 AIH의 1,2의 여러 ​​측면을 반영합니다. 또한, 유사한 에피토프의 특이성과 AIH-2 환자에서 발견 LKM-1 항체에 13 확산과 높은 titer 안티 CYP2D6 항체의 존재는 사전을 나타냅니다일반적인 만성자가 면역 반응의 sence. CYP2D6는 AIH-2의 주요 항원 있지만 더 자주 1 형 AIH 진단을 환자가 인식되지 않습니다. 또한, 그러한 기본 담즙 간경변 (PBC) 또는 기본 sclerosing cholangitis (PSC)와 같은 면역 병인 다른 간 질환에서 고통받는 환자가 특징의 autoantibodies을 생성하는 고유한 간 autoantigens과 그들의 간을 대한 특이성과 소형을 중심으로 서로 다른 병리 학적 특징 (PBC을 보여 ) 또는 대형 (PSC) 담즙 관. 그럼에도 불구하고, CYP2D6 모델 hepatocytes의 면역 파괴를 운전하고 질병을 치료할 수 치료 개입을 평가하는 핵심 선수를 식별할 수 있도록,자가 면역 질환 간 질환 중에 발생으로 만성 간장 염증 과정에 관여 immunopathogenic 메커니즘을 분석할 수있는 기회를 제공합니다.

<table border="1"><tbody><tr><td> 장비의 명칭 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 23G 바늘 </td><td> BD Biosciences </td><td> 300,800 </td><td></td></tr><tr><td> 27 ½ G 바늘 </td><td> VWR </td><td> 612-0151 </td><td></td></tr><tr><td> 30 ½ G 바늘 </td><td> BD Biosciences </td><td> 304,000 </td><td></td></tr><tr><td> 알라닌 아미노 전이 효소 테스트 스트립 (GPT / ALT) </td><td> 로슈 진단 </td><td> 10 745 138 202 </td><td></td></tr><tr><td> Aspartate 아미노 전이 효소 테스트 스트립 (알겠습니다 / 대서양 표준시) </td><td> 로슈 진단 </td><td> 10 745 120 202 </td><td></td></tr><tr><td> Anaesthesia 단위 Univentor 400 </td><td> <a href="http://www.agnthos.se" target="_blank" >AgnTho의</a> </td><td></td><td></td></tr><tr><td> 자료 금형, 일회용 37x24x10 mm의 </td><td> VWR </td><td> 720-0208 </td><td></td></tr><tr><td> 셀 스트레이너 70mm </td><td> VWR </td><td> 734-0003 </td><td></td></tr><tr><td> 그라 이오 스탯 </td><td> Leica </td><td> CM1850 자외선 </td><td></td></tr><tr><td> Heparinized 모세관 튜브 </td><td> 어부 </td><td> 3,123,987 </td><td></td></tr><tr><td> 현미경 Superfrost 플러스 슬라이드 </td><td> Menzel Gläser </td><td> J1800AMN​​Z </td><td></td></tr><tr><td> Microtainer SST 튜브 </td><td> BD Biosciences </td><td> 365,951 </td><td></td></tr><tr><td> Microtiter 번호판, V-바닥 </td><td> VWR </td><td> 391-1924 </td><td></td></tr><tr><td> Reflotron 플러스 </td><td> 로슈 진단 </td><td></td><td> 혈청 대서양 표준시 / ALT의 Determiniation </td></tr><tr><td colspan="4"></td></tr><tr><td> O의 이름을 지정F 시약 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 토끼 항 마우스 방지 콜라겐 유형 I IgG, </td><td> Chemicon </td><td> AB765P </td><td></td></tr><tr><td> Biotinylated 염소 항 토끼 IgG </td><td> 벡터 (AXXORA) </td><td> VC-BA-1000-MC15 </td><td></td></tr><tr><td> FITC-복합 백신 마우스 CD8a 항체 </td><td> 남부 생명 공학 </td><td> 1550-02 </td><td></td></tr><tr><td> PE-복합 백신 마우스 IFNγ 항체 </td><td> BD Biosciences </td><td> 554,412 </td><td></td></tr><tr><td> PE-복합 백신 마우스 CD16/CD32 항체 (FcR 블록) </td><td> BD Biosciences </td><td> 553,141 </td><td></td></tr><tr><td> Aquatex </td><td> VWR </td><td> 1.08562.0050 </td><td></td></tr><tr><td> 아비딘 / 비오틴 차단 키트 </td><td&gt; 벡터 (AXXORA) </td><td> VC-SP-2001-KI01 </td><td></td></tr><tr><td> 를 Brefeldin </td><td> 시그마 </td><td> B6542 </td><td></td></tr><tr><td> Collagenase IV </td><td> 시그마 </td><td> C5138-100MG </td><td></td></tr><tr><td> DAB 기판 키트 3&#39;3&#39;diaminobenzidine </td><td> 벡터 (AXXORA) </td><td> VC-SK-4100-KI01 </td><td></td></tr><tr><td> DNase </td><td> 시그마 </td><td> DN-25 </td><td></td></tr><tr><td> 엘리트 ABC 시약 </td><td> 벡터 (AXXORA) </td><td> VC-PK-7100-L050 </td><td></td></tr><tr><td> Eosin G / Y 솔루션 </td><td> 로스 </td><td> X883.1 </td><td></td></tr><tr><td> 태아 송아지 혈청 (FCS) </td><td> Biochrom AG </td><td> S 0115 </td><td></td></tr><tr><td> Isofluran </td><td> Forene </td><td> B506 </td><td></td></tr><tr><td> 메이어의 Hematoxylin 용액 </td><td> AppliChem </td><td> A4840, 1000 </td><td></td></tr><tr><td> 10월 운드 </td><td> 사쿠라 Finetek 유럽 </td><td> 4583 </td><td></td></tr><tr><td> PBS 버퍼 포름 알데히드 10% </td><td> 로스 </td><td> A146.3 </td><td></td></tr><tr><td> Percoll </td><td> Amersham </td><td> 17-0891-01 </td><td></td></tr><tr><td> Roti-Histokit </td><td> 로스 </td><td> 6638.1 </td><td></td></tr><tr><td> RPMI </td><td> Invitrogen </td><td> 61,870 </td><td></td></tr><tr><td> quillaja 껍질에서 사포닌 </td><td> 시그마 </td><td> S7900 </td><td></td></tr></tbody></table>

the cyp2d6 animal model: how to induce autoimmune hepatitis in mice

자가 면역 간염은 알 수없는 병인 1,2의 간을 드물지만 생명에 지장 면역 질환이다. 과거 수많은 시도에서 인간의 질병 3-5의 특성을 반영 동물 모델을 생성되었습니다. 그러나 다양한 모델로 질병의 유도 오히려 복잡했고 종종 간염 3-5에만 일시적했습니다. 따라서 트리거 6으로 제 2 형자가 면역 간염 (AIH-2), 즉 hCYP2D6에서 주요 인간 autoantigen를 사용하여 간단한 마우스 모델을 개발했습니다. hCYP2D6을 인식 1 형 간 - 신장 microsomal 항체 (LKM-1) 항체는 AIH-2 7,8의 특징입니다. wildtype FVB 또는 C57BL / 6 마​​우스에 hCYP2D6 게재가 간으로 트리거링 항원의 직접적인 전달을 보장합니다 아데노 바이러스 구조 (AD-2D6)에 의해했습니다. 따라서, 다음의 지역 염증이 autoimmunity의 후속 개발을위한 비옥한 분야 9를 생성합니다. C광고-2D6의 정맥 주사와 intraperitoneal 주사의 ombination은 간장에 대한 지속적인 면역 손상 (섹션 1) 유도하기위한 가장 효과적인 경로이다. 여기 간 질환은 CYP2D6 모델에서 유도되는 방법과 간 손상의 다양한 측면을 평가하는 방법을자가 면역 질환에 대한 상세한 프로토콜을 제공합니다. 첫째, hepatocyte 같은 aminotransferases 등의 파괴,뿐만 아니라 hCYP2D6 항체의 titers을 나타내는 마커의 혈청 수준은 혈액 retroorbitaly (섹션 2) 샘플링에 의해 결정됩니다. 둘째, hCYP2D6 특정 T 세포 반응은 비장과 간장에서 lymphocytes를 수집하는이 특징입니다. 순수 간 lymphocytes를 얻기 위해, 간은는 콜라겐으로 소화와 Percoll 기울기 (섹션 4) 위에 정화 포털 정맥 (섹션 3)를 통해 PBS에 의해 perfused됩니다. hCYP2D6 특정 T 세포의 빈도는 hCYP2D6 펩티드 및 유동세포계측법 의한 IFNγ 생산 세포 (섹션 5)의 식별과 자극에 의​​해 분석됩니다. 셋째,세포 침투와 섬유증은 간암 섹션 (섹션 6) immunohistochemistry에 의해 결정됩니다. 이러한 분석 섭생이 모델의 만성적인 성격을 증명하기 위해서는 질병의 개시 이후에 여러 번 실시되어야한다. 주파수와 활동 hCYP2D6 특정 T 및 / 또는 B 세포 및 간 손상과 섬유증의 정도 특징으로 면역 반응의 정도는 예방 지연이나 autodestructive을 폐기하기위한 가능한 치료의 후속 평가를 위해 평가되어야 간장의 과정입니다.

Watch this Scientific Journal Video about The CYP2D6 Animal Model: How to Induce Autoimmune Hepatitis in Mice at JoVE.com

The CYP2D6 Animal Model: How to Induce Autoimmune Hepatitis in Mice

주요 인간 autoantigen을 표현 아데노 바이러스와 생쥐의 감염은 CYP2D6의 광범 간염, 섬유증 및 생성을 특징으로 면역 - 매개 간 질환의 영구적인 형태로 제 2 형자가 면역 간염 결과에서 고통받는 환자 세라 인정 P450 2D6 (hCYP2D6)를 시토크롬 특정 면역 반응.

CYP2D6 동물 모델 : 마우스에 면역 간염을 유발하는 방법

Autoimmune hepatitis is a rare but life threatening autoimmune disease of the liver of unknown etiology1,2. In the past many attempts have been made to ...

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18.9K 조회수.  Goethe University Hospital Frankfurt. 주요 인간 autoantigen을 표현 아데노 바이러스와 생쥐의 감염은 CYP2D6의 광범 간염, 섬유증 및 생성을 특징으로 면역 - 매개 간 질환의 영구적인 형태로 제 2 형자가 면역 간염 결과에서 고통받는 환자 세라 인정 P450 2D6 (hCYP2D6)를 시토크롬 특정 면역 반응.

비디오: The CYP2D6 Animal Model: How to Induce Autoimmune Hepatitis in Mice

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged mechanical ventilation, contributes to morbidity and mortality of critical illness, and studies on novel genetic or pharmacological targets are areas of intense investigation (1-3, 5, 8, 26, 30, 33-36). ALI is defined by the acute onset of the disease, which leads to non-cardiac pulmonary edema and subsequent impairment of pulmonary gas exchange (36). We have developed a murine model of ALI by using a pressure-controlled ventilation to induce ventilator-induced lung injury (2). For this purpose, C57BL/6 mice are anesthetized and a tracheotomy is performed followed by induction of ALI via mechanical ventilation. Mice are ventilated in a pressure-controlled setting with an inspiratory peak pressure of 45 mbar over 1 - 3 hours. As outcome parameters, pulmonary edema (wet-to-dry ratio), bronchoalveolar fluid albumin content, bronchoalveolar fluid and pulmonary tissue myeloperoxidase content and pulmonary gas exchange are assessed (2). Using this technique we could show that it sufficiently induces acute lung inflammation and can distinguish between different treatment groups or genotypes (1-3, 5). Therefore this technique may be helpful for researchers who pursue molecular mechanisms involved in ALI using a genetic approach in mice with gene-targeted deletion.

A murine model for ventilator induced lung injury is an important tool to study an acute lung injury in vivo. Here, we report an easy applicable in situ model for acute lung injury using high-pressure mechanical ventilation to induce acute failure of the lung.

마우스의 급성 폐 상해를 유발하는 압력 제어 환기

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

연구

의학

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

PTSD의, 신경 Hematologic, 그리고 행동 상관 관계를 누설하는 행위에 대해서는 동물 모델에서 생체

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

마우스의 긴 기간 온 몸이 고열에 대한 프로토콜

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

소설 허벌 요법을 평가하는 패혈증의 동물 모델의 사용

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of the hepatic blood supply is poorly oxygenated portal vein blood derived from the gastrointestinal tract and spleen. Oxygen is delivered to hepatocytes by blood flowing from a terminal branch of the portal vein to a central venule via sinusoids, and this makes an oxygen gradient in hepatic lobules. The oxygen gradient is an important physical parameter that involves the expression of enzymes upstream and downstream in hepatic microcirculation, but the lack of techniques for measuring oxygen consumption in the hepatic microcirculation has delayed the elucidation of mechanisms relating to oxygen metabolism in liver. We therefore used FITC-labeled erythrocytes to visualize the hepatic microcirculation and used laser-assisted phosphorimetry to measure the partial pressure of oxygen in the microvessels there. Noncontact and continuous optical measurement can quantify blood flow velocities, vessel diameters, and oxygen gradients related to oxygen consumption in the liver. In an acute hepatitis model we made by administering acetaminophen to mice we observed increased oxygen pressure in both portal and central venules but a decreased oxygen gradient in the sinusoids, indicating that hepatocyte necrosis in the pericentral zone could shift the oxygen pressure up and affect enzyme expression in the periportal zone. In conclusion, our optical methods for measuring hepatic hemodynamics and oxygen consumption can reveal mechanisms related to hepatic disease.

An optical system was developed to visualize hepatic microcirculation with FITC-labeled erythrocytes and to measure the partial pressure of oxygen in the microvessels with laser-assisted phosphorimetry. This method can be used to investigate physiological and pathological mechanisms by analyzing microvascular structure, diameter, blood flow velocity, and oxygen tension.

급성 간염 모델 신청 : 시각화와 간장 Microcirculation의 혈류와 산소 소비 분석

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of ...

Implanting Glass Spinal Cord Windows in Adult Mice with Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as multiple sclerosis. Here we present a low-cost and reproducible glass window implantation protocol that is suitable for intravital microscopy and studying the dynamics of spinal cord cytoarchitecture with subcellular resolution in live adult mice with EAE. Briefly, we surgically expose the vertebrae T12-L2 and construct a chamber around the exposed vertebrae using a combination of cyanoacrylate and dental cement. A laminectomy is performed from T13 to L1, and a thin layer of transparent silicone elastomer is applied to the dorsal surface of the exposed spinal cord. A modified glass cover slip is implanted over the exposed spinal cord taking care that the glass does not directly contact the spinal cord. To reduce the infiltration of inflammatory cells between the window and spinal cord, anti-inflammatory treatment is administered every 2 days (as recommended by ethics committee) for the first 10 days after implantation. EAE is induced only 2-3 weeks after the cessation of anti-inflammatory treatment.
Using this approach we successfully induced EAE in 87% of animals with implanted windows and, using Thy1-CFP-23 mice (blue axons in dorsal spinal cord), quantified axonal loss throughout EAE progression. Taken together, this protocol may be useful for studying the recruitment of various cell populations as well as their interaction dynamics, with subcellular resolution and for extended periods of time. This intravital imaging modality represents a valuable tool for developing therapeutic strategies to treat autoimmune demyelinating diseases such as EAE.

We describe a method for implanting and maintaining glass windows over the exposed spinal cords of adult mice for studying experimental autoimmune encephalomyelitis. These windows provide chronic optical access to the spinal cord for monitoring cell dynamics at the subcellular level in live animals using two-photon microscopy.

실험자가 면역 뇌척수염 성인 마우스에 유리 척수 Windows를 이식

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as ...

A Magnetic Microbead Occlusion Model to Induce Ocular Hypertension-Dependent Glaucoma in Mice

The use of rodent models of glaucoma has been essential to understand the molecular mechanisms that underlie the pathophysiology of this multifactorial neurodegenerative disease. With the advent of numerous transgenic mouse lines, there is increasing interest in inducible murine models of ocular hypertension. Here, we present an occlusion model of glaucoma based on the injection of magnetic microbeads into the anterior chamber of the eye using a modified microneedle with a facetted bevel. The magnetic microbeads are attracted to the iridocorneal angle using a handheld magnet to block the drainage of aqueous humour from the anterior chamber. This disruption in aqueous dynamics results in a steady elevation of intraocular pressure, which subsequently leads to the loss of retinal ganglion cells, as observed in human glaucoma patients. The microbead occlusion model presented in this manuscript is simple compared to other inducible models of glaucoma and also highly effective and reproducible. Importantly, the modifications presented here minimize common issues that often arise in occlusion models. First, the use of a bevelled glass microneedle prevents backflow of microbeads and ensures that minimal damage occurs to the cornea during the injection, thus reducing injury-related effects. Second, the use of magnetic microbeads ensures the ability to attract most beads to the iridocorneal angle, effectively reducing the number of beads floating in the anterior chamber avoiding contact with other structures (e.g., iris, lens). Lastly, the use of a handheld magnet allows flexibility when handling the small mouse eye to efficiently direct the magnetic microbeads and ensure that there is little reflux of the microbeads from the eye when the microneedle is withdrawn. In summary, the microbead occlusion mouse model presented here is a powerful investigative tool to study neurodegenerative changes that occur during the onset and progression of glaucoma.

Here, we present a protocol to induce ocular hypertension in the murine eye that results in the loss of retinal ganglion cells as observed in glaucoma. Magnetic microbeads are injected into the anterior chamber and attracted to the iridocorneal angle using a magnet to block the outflow of aqueous humour.

자기 마이크로 비드 폐색 모델 마우스의 녹내장 안구 고혈압 종속 유도합니다

The use of rodent models of glaucoma has been essential to understand the molecular mechanisms that underlie the pathophysiology of this multifactorial ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

풍선-기반 부상 마우스 복 부 대동맥에서 Myointimal 증식을 유도 하

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

마우스에 가로 대동맥 수축 유도를 폐쇄 가슴 모델

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...