The authors acknowledge the following funding sources, Whittier Foundation, Harrington Foundation, National Institutes of Health and Research to Prevent Blindness.

모든 탈핵 눈은 도살장에서 얻은하고 모든 실험은 기관 바이오 안전성 법에 따라 수행되었다. <ol><li> 표면에 세포핵이 제거 된 눈을 고정합니다. 눈 주위에 여분의 조직을 통해 조직의 핀을 배치하고 스티로폼 보드로를 확보하여이 작업을 수행합니다. </li><li> 해부 및 눈에서 perilimbal 결막을 분리합니다. <ol><li> 윤부에서 결막을 절개하고 퉁명스럽게 공막 떨어져 그것을 해부 미세 집게 (0.3 집게) 및 microscissors (웨스트 코트 위)를 사용합니다. 무딘 절개가 더 해부 할 수 있도록 진행으로 윤부 따라 결막을 잘라. </li><li> 가능한 한 많은 공막을 노출 끝까지 눈 (360도)의 주위에 결막을 제거합니다. 참고 :이 절차의 나머지 부분 동안 수술 핀이나 집게로 눈에 집착 용이하게하기 위해 결막의 작은 금액을 떠나 도움이됩니다. </li></ol></li><li> 하나 따라 전체 두께 공막 편 만들기눈 (메스 블레이드 69)의 측면. <ol><li> 부드럽게 어둡게 착색 된 맥락막까지 메스 (메스 블레이드 11)과 공막으로 절단하여 윤부에 윤부 및 3mm 후방에 ~ 5mm 공막 절개 평행을 볼 수 있습니다. 맥락막 자체를 절개하지 않도록주의하십시오. </li><li> 이 부드럽게 안쪽으로 맥​​락막을 눌러이 조직 사이의 잠재적 인 공간을 확대 할 수있을 때까지 조심스럽게, (무딘 절개를 사용하여) 가위 또는 메스 중, 공막과 맥락막 사이의 평면을 따라 해부. </li><li> T 자형 절개를 작성, 날카로운 microscissors와 최초의 공막 절개에 수직 다른 공막 절개를합니다. </li><li> 그리고 퉁명스럽게 기본 맥락막에서 공막 조직을 제거하고 부드럽게 위에서 언급 한 바와 같이 공막에서 멀리 맥락막을 밀어 원주 방식으로 해부하는 것을 계속한다. </li><li> 필요에 따라 공막 편을 크게한다. <ol><li> 부드럽게 떨어져 공막에서 맥락막을 해부 무딘과정 전반에 걸쳐. </li><li> 윤부 수직 만들어진 절개 시신경 도달 절개 윤부 평행이 눈 둘레 (45)의 적어도 1/3까지 이루어 플랩을 크게한다. </li><li> 무딘 해부하는 동안 눈을 조작하기위한 지렛대의 소스로 공막 편을 사용합니다. 플랩에 반대 견인은 무딘 절개보다 쉽게​​ 만든다. </li></ol></li><li> 다른 공막 편에 이전 단계를 반복합니다. </li></ol></li><li> 맥락막의 큰 영역을 노출하는 공막 플랩을 버려야. </li><li> 눈의 둘레 (360도)의 주위에 3 단계에서 만든 절개를 계속합니다. </li><li> 나머지 맥락막 - 망막 조직을 제거합니다. <ol><li> 노출 된 지역 내에서 조심스럽게 남아있는 맥락막 - 망막 조직을 브러시면 팁을 사용합니다. 또한, 부드럽게 집게로 조직을 잡고 기본 유리체 오프를 껍질. </li><li> 필요하다면 맥락막 성으로 절개를시신경에서 arting. 그런 다음, 망막 및 유리체 피질을 노출 부드럽게 맥락막을 벗겨. </li></ol></li><li> 멀리 공막에서 맥락막을 해부 무딘 계속하고 전체 그대로 유리체를 얻기 위해 맥락막을 벗겨. </li><li> 원하는 위치에서 전체 유리체의 위치를​​ 각막에 부착 된 공막 림을 사용합니다. 이 시점에서, 유리체는 전방 및 공막 렌즈에 부착된다. </li><li> 모든 공막을 제거, 렌즈 주위 공막의 내부로부터 해부 무딘 유리체. </li><li> 필요가있을 경우 멀리 유리체에서 렌즈를 특종 무딘 도구를 사용합니다. 다양한 vitreodynamic 실험 샘플을 사용합니다. <ol><li> 공기에 노출 알려진 표면적 유리 비커에 샘플을 놓습니다. 샘플의 가장자리에 산소에 민감한 프로브를 놓습니다. 샘플로 알려진 거리 (R)에 프로브를 이동하는 마이크로 매니퓰레이터를 사용합니다. </li><li> 이론적 확산 방정식을 얻기 위해 확산의 픽의 법칙을 사용합니다. 다 함께TA는 단계 10.1에 수집 등 실험적 확산 계수 / 반응 조건을 구하는 </li></ol></li></ol>

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The authors have nothing to disclose.

조심스럽게 유리체 박리 동안 수행해야 두 가지 중요한 단계가 있습니다. 전체 두께 공막 편을 만드는 단계 3, 전체 박리에 결정적이다. 케어는 전체 두께 공막 편을 만들 때 맥락막으로 절단하지 않도록주의해야한다. 다른 중요한 단계는 맥락막에서 공막을 멀리 해부한다. 이 단계는주의 깊게 유리체가 밖으로 유출 할 수있는 맥락막에 여러 개의 구멍을 만드는 방지하기 위해 수행해야합니다. 프...

이 방법의 목적은 구체적으로 vitreodynamic 분석의 목적을 위해, 사체로부터 눈, 유리질 코어 및 피질 손상으로, 전체 그대로 유리체를 분리하는 기술이다. 유리체 생리학 분야는 유체 역학 연구원 등, 다양한 분야 연구자, 성장함에 따라, 유리체 1의 물리적, 역학적 특성을 조사하고 있습니다. 이를 위해, 그것은 세부 사항에 다양한 분야의 연구자를 지원하기 위해 전체 그대로 유리체를 분리하는 기술이 필수적이다. Sebag 등. 2 등 (3) 인간의 시체 눈에 우아한 전체 유리체 해부를 수행하고 그 결과의 그림을 보여 주었다. 그러나, 사용되는 기술은 독립적으로 복제하는 방법을 할 수 없을 것이다 디테일 비전문가에 설명되지 않았다. 다른 연구는 이러한 포부 또는 부분 절개와 같은 간단한 방법을 사용하여 시체의 눈에서 유리체 수확 한그 중 둘은 전체 그대로 유리체가 발생하지 않습니다. Gisladottis 등. 4, 쑤 외. (5)는 시체의 눈에서 수확 유리체 유머의 투과성을 조사합니다. 유리체 추출 할 수있는 방법을 설명하지 않았다 때문에, 그것은 그들이 주사기 유리체를 흡입하는 것으로 가정 하였다. 와트 등. 6은 수술 기법과 토끼 유리체 유머를 분리하는 방법을 설명함으로써 한 단계 더 갔다. 그러나,이 방법은 유리질 코어의 단리없는 유리체 피질 초래한다. Skeie 등. (7)는 고유 한 후에 4 개의 영역으로 편성 유리체 우아 분석을 위해 각각의 부분을 절개하는 방법을 설명했다. 그러나이 기술은, 전체로서 유리체 그대로 발생하지 않는다. 현재의 기술은 현재 사체의 눈을 수행하는 생물 물리학 적 실험을 용이하게하기 위해 개발되었다. 이전 방법으로 설명보브는, 1) 없음이 완전히 전체 유리체를 분리하지 않기 때문에 제한됩니다, 2) 수확 유리체 코어 피질 3) 유리체 해부학 적 구조가 유지되지 않습니다, 균질화, 또는 4) 해부 기술은 적절하게 다른 분야의 연구자들에 의해 복제에 대해 자세히 설명하지 않습니다 . 또한 공막과 맥락막, 유리체의 가시화에 의한 불투명도가 그대로 안구 제한된다. 이것은 전체 눈 안에 이루어질 수 측정의 정밀도 및 가능성을 제한한다. 또한, 유리체 주변 해부학 적 구조는 유리질의 생화학 적 및 물리적 특성의 연구를 혼동 할 수있다. 최근 몇 년 동안, 유리체 과학의 몸은 엄청나게 성장하고 전체 유리체는 개별 부품과는 다른 특성을 가지고 있다고 생각하는 이유가있다. vitreodynamics의 researc의 유리체의 물리적, 역학적, 화학적 특성을 조사에 관심이 증가하고있다이러한 약물 전달, 체내 산소 (8)과 유리체 절제술 임상 의학에서 응용 프로그램이 시간. 유리체를 조작 약리학 제제를 사용 약리 vitreodynamics는, 유리체 결과 9을 향상 시키는데 사용될 수있다. 생체 역학적 특성은 유리 체내 약물 전달 기술 10-12을 개선하는데 사용될 수있다 유리질 유체의 흐름을 모델링하는 데 사용된다. 유리체의 다양한 세그먼트의 물리적 특성은 vitreo - 망막 산소 수송 (13)을 이해하는 데 매우 중요하다. 제안 된 유리체 박리 기술은 그대로 유리체 유머의 다양한 속성을 연구하는 데 사용할 수 있습니다. 이 벤치 탑 실험이 더 나은 시각화 전체 그대로 유리체 기관에서 수행 할 수 있습니다. 요약하면, 유리체의 연구를위한 현재의 방법은 어느 적절히 설명을 생략하거나, 불완전한 절연 및 유리질 코어 피질 초래한다. 따라서, 예를 수행 할 필요가있다투명한 눈 모델 xperiments은 시체의 눈에 존재하는 유리체의 해부학을 유지하면서.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>0.3 forceps</td>
			<td>Storz Opthalmics</td>
			<td>E1793</td>
			<td></td>
		</tr>
		<tr>
			<td>Westcott Tenotomy Scissors Curved Right</td>
			<td>Storz Opthalmics</td>
			<td>E3320 R</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Handle No. 3</td>
			<td>VWR</td>
			<td>25607-947</td>
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		<tr>
			<td>Scalpel Blade, #11, for #3 Handle</td>
			<td>VWR</td>
			<td>470174-844</td>
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whole vitreous humor dissection for vitreodynamic analysis

The authors propose an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes. While previous studies have shown the results of such dissections, the detailed steps have not been described, precluding researchers outside the field from replicating their methods. Other studies harvest vitreous either through aspiration, which does not maintain the vitreous structure anatomy, or through partial dissection, which only isolates the vitreous core. The proposed method isolates the whole vitreous body, with the vitreous core and cortex intact, while maintaining vitreous anatomy and structural integrity. In this method, a full thickness scleral flap in an enucleated porcine eye is first created and through this, the choroid tissue can be separated from the sclera. The scleral flap is then expanded and the choroid is completely separated from the sclera. Finally the choroid-retina tissue is peeled off the vitreous to leave an isolated intact vitreous body. The proposed vitreous dissection technique can be used to study physical properties of the vitreous humor. In particular, this method has significance for experimental studies involving drug delivery, vitreo-retinal oxygen transport, and intraocular convection.

프로토콜을 따르는 것은 그대로 코어와 피질 (그림 3) 성공적인 유리체 박리로 이어질 것입니다. 이 유리체 피질에 부착 망막의 잔여 부분으로부터 명백하다. 본래 전체 유리체 특정 vitreodynamic 실험을위한 여러 가지 방법으로 사용될 수있다. 우리의 경우에는, 확산 유리체 그대로 산소의 속도와 그 대응하는 시간 상수 (도 2)의 조사 하였다. 우리의 방법을 사용하여 (코어 ?...

Watch this Scientific Journal Video about Whole Vitreous Humor Dissection for Vitreodynamic Analysis at JoVE.com

Whole Vitreous Humor Dissection for Vitreodynamic Analysis

The goal of this protocol is to show an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes.

Vitreodynamic 분석을위한 항목의 유리체 유머 해부

The authors propose an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes. While previous ...

whole-vitreous-humor-dissection-for-vitreodynamic-analysis

Research

JoVE Journal

Medicine

11.3K 조회수.  University of Southern California. The goal of this protocol is to show an effective technique to isolate whole, intact vitreous core and cortex from post mortem enucleated porcine eyes.

비디오: Whole Vitreous Humor Dissection for Vitreodynamic Analysis

Dissection of Human Vitreous Body Elements for Proteomic Analysis

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.

This video shows an effective technique for differentiating and dissecting the various semi-transparent structures of the human vitreous body in post mortem eyes.

Proteomic 분석을위한 인적 유리 바디 요소의 해부

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions ...

연구

의학

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 ...

3D Whole-heart Myocardial Tissue Analysis

Cardiac regenerative therapies aim to protect and repair the injured heart in patients with ischemic heart disease. By injecting stem cells or other biologicals that enhance angio- or vasculogenesis into the infarct border zone (IBZ), tissue perfusion is improved, and the myocardium can be protected from further damage. For maximum therapeutic effect, it is hypothesized that the regenerative substance is best delivered to the IBZ. This requires accurate injections and has led to the development of new injection techniques. To validate these new techniques, we have designed a validation protocol based on myocardial tissue analysis. This protocol includes whole-heart myocardial tissue processing that enables detailed two-dimensional (2D) and three-dimensional (3D) analysis of the cardiac anatomy and intramyocardial injections. In a pig, myocardial infarction was created by a 90-min occlusion of the left anterior descending coronary artery. Four weeks later, a mixture of a hydrogel with superparamagnetic iron oxide particles (SPIOs) and fluorescent beads was injected in the IBZ using a minimally-invasive endocardial approach. 1 h after the injection procedure, the pig was euthanized, and the heart was excised and embedded in agarose (agar). After the solidification of the agar, magnetic resonance imaging (MRI), slicing of the heart, and fluorescence imaging were performed. After image post-processing, 3D analysis was performed to assess the IBZ targeting accuracy. This protocol provides a structured and reproducible method for the assessment of the targeting accuracy of intramyocardial injections into the IBZ. The protocol can be easily used when the processing of scar tissue and/or validation of the injection accuracy of the whole heart is desired.

This protocol describes a novel method for the 3D comparison of whole-heart myocardial tissue with MRI. This is designed for the accurate assessment of intramyocardial injections in the infarct border zone of a chronic porcine model of myocardial infarction.

3D 전체 심장 심근 조직 분석

Cardiac regenerative therapies aim to protect and repair the injured heart in patients with ischemic heart disease. By injecting stem cells or other ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at the 12 codon (Ptf1acre/+;LSL-KrasG12D/+) mouse strain as a model of human pancreatic cancer. The goal of our current studies is to identify novel metabolic biomarkers of pancreatic cancer progression. We have performed metabolic profiling of urine, feces, blood, and pancreas tissue extracts, as well as histological analyses of the pancreas to stage the cancer progression. The mouse pancreas is not a well-defined solid organ like in humans, but rather is a diffusely distributed soft tissue that is not easily identified by individuals unfamiliar with mouse internal anatomy or by individuals that have little or no experience performing mouse organ dissections. The purpose of this article is to provide a detailed step-wise visual demonstration to guide novices in the removal of the mouse pancreas by dissection. This article should be especially valuable to students and investigators new to research that requires harvesting of the mouse pancreas by dissection for metabolic profiling or histological analyses.

This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.

조직학 분석 및 대사 프로 파일링에 대 한 마우스 췌의 해 부

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

Dissection and Explant Culture of Murine Allantois for the In Vitro Analysis of Allantoic Attachment

The placenta is essential for the growth and development of mammalian embryos. For this reason, numerous genetic alterations and likely also environmental insults that disturb placenta development or function can cause early pregnancy loss in mice and humans. Nevertheless, simple in vitro assays to screen for potential effects on placenta formation are lacking. Here, we focus on modeling the first and critical step in placenta formation, which consists of the attachment of the allantois to the chorion. We describe a method to rapidly assess the attachment of allantoic explants on immobilized &#945;4&#946;1 integrin, which serves as a chorio-mimetic substrate.This in vitro approach enables a qualitative evaluation of the attachment and spreading behavior of multiple allantois explants at different consecutive time points. The protocol may be used to investigate the effect of targeted mouse mutations, drugs, or various environmental factors that have been linked to pregnancy complications or fetal loss on allantois attachment ex vivo.

Dissection and Explant Culture of Murine Allantois for the In Vitro Analysis of Allantoic Attachment

We describe an in vitro assay to model chorioallantoic attachment, the first step in placenta formation. The protocol demonstrates the dissection and explant culture of murine allantoides on immobilized &#945;4&#946;1 integrin. Allantois attachment is evaluated microscopically at pre-determined time points.

해 부와 Allantoic 첨부 파일의 생체 외에서 분석 Murine Allantois Explant 문화

The placenta is essential for the growth and development of mammalian embryos. For this reason, numerous genetic alterations and likely also environmental ...

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&#8212;Application in Premanifest Huntington's Disease

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy begins many years prior to the motor onset (during the &#34;premanifest&#34; period), but the spatiotemporal pattern of regional atrophy across the brain has not been fully characterized. Here we demonstrate an online cloud-computing platform, &#34;MRICloud&#34;, which provides atlas-based whole-brain segmentation of T1-weighted images at multiple granularity levels, and thereby, enables us to access the regional features of brain anatomy. We then describe a regression model that detects statistically significant inflection points, at which regional brain atrophy starts to be noticeable, i.e. the &#34;change-point&#34;, with respect to a disease progression index. We used the CAG-age product (CAP) score to index the disease progression in HD patients. Change-point analysis of the volumetric measurements from the segmentation pipeline, therefore, provides important information of the order and pattern of structural atrophy across the brain. The paper illustrates the use of these techniques on T1-weighted MRI data of premanifest HD subjects from a large multicenter PREDICT-HD study. This design potentially has wide applications in a range of neurodegenerative diseases to investigate the dynamic changes of brain anatomy.

Whole-brain Segmentation and Change-point Analysis of Anatomical Brain MRI&amp;#8212;Application in Premanifest Huntington&#039;s Disease

This paper describes a statistical model for volumetric MRI data analysis, which identifies the &#34;change-point&#34; when brain atrophy begins in premanifest Huntington&#39;s disease. Whole-brain mapping of the change-points is achieved based on brain volumes obtained using an atlas-based segmentation pipeline of T1-weighted images.

전체 두뇌 세분화 및 해 부 뇌 MRI의 변화 포인트 분석-Premanifest Huntington의 질병에 응용

Recent advances in MRI offer a variety of useful markers to identify neurodegenerative diseases. In Huntington&#39;s disease (HD), regional brain atrophy ...

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of aortic disease exist, in vivo imaging of the vasculature has been limited. In recent years, micro-computerized tomography (micro-CT) has emerged as a preferred modality for imaging both large and small vessels both in vivo and ex vivo. In conjunction with a method of vascular casting, we have successfully used micro-CT to characterize the frequency and distribution of aortic pathology in &#946;-aminopropionitrile-treated C57/Bl6 mice. Technical limitations of this method include variations in the quality of the perfusion introduced by poor animal preparation, the application of proper methodologies for vessel size quantification, and the non-survivability of this procedure. This article details a methodology for the intravascular perfusion of a lead-based radiopaque silicone rubber for the quantitative characterization of aortopathy in a mouse model of aneurysm and dissection. In addition to visualizing aortic pathology, this method may be used for examining other vascular beds in vivo or vascular beds removed post-mortem.

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &amp;#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

This article describes a detailed methodology of using a radiopaque lead-based silicone rubber to perfuse the murine vasculature for aortic diameter quantification in a mouse model of aortic aneurysm and dissection.

Β-aminopropionitrile-유도 대동맥 동맥 및 해 부의 마우스 모델에서 Aortopathy의 양적 마이크로 CT 분석

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...

Imaging and Analysis of Oil Red O-Stained Whole Aorta Lesions in an Aneurysm Hyperlipidemia Mouse Model

Apolipoprotein E (Apoe)- or low density lipoprotein receptor (Ldlr)-deficient hyperlipidemic mice are the two most commonly used models for atherosclerosis research. They are used to study the impact of a various genetic factors and different cell types on atherosclerotic lesion formation and as well as test the development of new therapies. Isolation, excision of the whole aorta, and quantification of Oil Red O-stained atherosclerotic lesions are basic morphometric methods used to evaluate atherosclerotic burden. The goal of this protocol is to describe an optimized, step-by-step surgical method to dissect, perfuse-fix, isolate, stain, image and analyze atherosclerotic lesions in mouse aortas with Oil Red O. Because atherosclerotic lesions can form anywhere in the entire aortic tree, this whole aorta Oil Red O staining method has the advantage of evaluating lipid-laden plaques in the entire aorta and all branches in a single mouse. In addition to Oil Red O staining, fresh isolated whole aortas can be used for variety of in vitro and in vivo experiments and cell isolations.

This protocol provides a step-by-step procedure to analyze atherosclerotic burden in mice. Investigators can use this protocol to compare the abundance, location, and size of atherosclerotic lesions in different animals.

Apolipoprotein E (Apoe)- or low density lipoprotein receptor (Ldlr)-deficient hyperlipidemic mice are the two most commonly used models for ...

Location, Dissection, and Analysis of the Murine Stellate Ganglion

The autonomic nervous system is a substantial driver of cardiac electrophysiology. Especially&#160;the role of its sympathetic branch is an ongoing matter of investigation in the pathophysiology of ventricular arrhythmias (VA). Neurons in the stellate ganglia (SG)&#160;&#8211;&#160;bilateral star-shaped structures of the sympathetic chain &#8211;&#160;are an important component of the sympathetic infrastructure. The SG are a recognized target for&#160;treatment via cardiac sympathetic denervation in patients with therapy-refractory VA. While neuronal remodeling and glial activation in the SG have been described in patients with VA, the underlying cellular and molecular processes that potentially precede the onset of arrhythmia are only insufficiently understood and should be elucidated to improve autonomic modulation. Mouse models allow us to study sympathetic neuronal remodeling, but identification of the murine SG is challenging for the inexperienced investigator. Thus, in-depth cellular and molecular biological studies of the murine SG are lacking for many common cardiac diseases. Here, we describe a basic repertoire for dissecting and studying the SG in adult mice for analyses at RNA level (RNA isolation for gene expression analyses, in situ hybridization), protein level (immunofluorescent whole mount staining), and cellular level (basic morphology, cell size measurement). We present potential solutions to overcome challenges in the preparation technique, and how to improve staining via quenching of autofluorescence. This allows for the visualization of neurons as well as glial cells via established markers in order to determine cell composition and remodeling processes. The methods presented here allow characterizing the SG to gain further information on autonomic dysfunction in mice prone to VA and can be complemented by additional techniques investigating neuronal and glial components of the autonomic nervous system in the heart.

Pathophysiological changes in the cardiac autonomic nervous system, especially in its sympathetic branch, contribute to the onset and maintenance of ventricular arrhythmias. In the present protocol, we show how to characterize murine stellate ganglia to improve the understanding of the underlying molecular and cellular processes.

뮤린 스텔레이트 갠리온의 위치, 해부 및 분석

The autonomic nervous system is a substantial driver of cardiac electrophysiology. Especially&#160;the role of its sympathetic branch is an ongoing matter ...

A Non-Invasive Method for Generating the Cyclic Loading-Induced Intra-Articular Cartilage Lesion Model of the Rat Knee

The pathophysiology of primary osteoarthritis (OA) remains unclear. However, a specific subclassification of OA in relatively younger age groups is likely correlated with a history of articular cartilage damage and ligament avulsion. Surgical animal models of OA of the knee play an important role in understanding the onset and progression of post-traumatic OA and aid in the development of novel therapies for this disease. However, non-surgical models have been recently considered to avoid traumatic inflammation that could affect the evaluation of the intervention. 
In this study, an intra-articular cartilage lesion rat model induced by in vivo cyclic compressive loading was developed, which allowed researchers to (1) determine the optimal magnitude, speed, and duration of load that could cause focal cartilage damage; (2) assess post-traumatic spatiotemporal pathological changes in chondrocyte vitality; and (3) evaluate the histological expression of destructive or protective molecules that are involved in the adaptation and repair mechanisms against joint compressive loads. This report describes the experimental protocol for this novel cartilage lesion in a rat model.

Here, we present the cyclic loading-induced intra-articular cartilage lesion model of the rat knee, generated by 60 cyclic compressions over 20 N, resulting in damage to the femoral condylar cartilage in rats.

래트 무릎의 순환 로딩 유도 관절내 연골 병변 모델을 생성하기 위한 비침습적 방법

The pathophysiology of primary osteoarthritis (OA) remains unclear. However, a specific subclassification of OA in relatively younger age groups is likely ...