为视力斗争提供了资金。组织获得来自爱荷华州狮子会眼库。

1。眼前段解剖。 <ol><li>角膜是首先在角膜缘切口使用15 °刀片（图1）进入前室，中删除。然后一个弧形角膜巩膜剪刀的刀片插入前房。圆周削减是在角膜上，角膜缘前。 </li><li> 0.12科利柏钳和韦斯科特剪刀用于切割虹膜睫状体周前。 </li><li>晶状体核与0.12科利柏钳或15 °刀片（supersharp）和钳皮层与胶囊被删除。 </li></ol> 2。玻璃体核心的愿望。 <ol><li>一个23号针头，5毫升注射器插入中期玻璃体（图1）。 </li><li>约1毫升或玻璃体核心是轻轻吸气。 </li><li>样本，然后放置在离心管中，再放入液态氮。 </li></ol> 3。前部玻璃体剥离。 <ol><li>通过调整入射光从鹅颈显微镜光前玻璃体被看作是一个半透明的戒指。前吸玻璃体核心分开便于识别的其他结构的前玻璃体。 </li><li>弹性组织的表仔细拉到远离科利柏或敷料钳睫状体和切Vannas剪刀。这一演习是重复的。 </li><li>样本，然后放置在离心管中，再放入液态氮。 </li></ol> 4。玻璃体基底部夹层。 <ol><li>使用0.12钳稳定眼罩，韦斯科特剪刀用“花”的眼睛，使四个等距的减压削减对视神经头睫状体。 </li><li>睫状体平坦（图2）蚌是从每个象限使用韦斯科特剪刀。 </li><li>玻璃体基底部，然后抓住钳在玻璃体和视网膜（图2）锯齿缘两侧。 </li><li>玻璃体基底部的持续牵引，应用镊子。韦斯科特剪刀连续切割提取“珍珠串”的外观，半透明的组织，因为它是切除。 </li><li>样本，然后放置在离心管中，再放入液态氮。 </li></ol> 5。玻璃体皮质去除。 <ol><li>韦斯科特剪刀每个象限切除玻璃体切割后的锯齿缘（图2）单张约3毫米。 </li><li>组织之间的传单，玻璃体皮质可视化的半透明膜。 </li><li> 0.12钳或弹性薄膜是抓住坚持一个Weck - CEL手术海绵举行。然后拉到远离视网膜玻璃体皮质韦斯科特剪刀（图1）切除。 </li><li>样品放置在离心管中，再放入液态氮。所有样本保存于-80℃直至实验利用。 </li></ol> 6。代表性的成果组织样本，可以处理各种具体的实验方法。在我们的例子中，样品进行蛋白质分析提交经SDS - PAGE（图3）。 <img src="/files/ftp_upload/2455/2455fig1.jpg" alt="图1" /> 图1。横断面人眼玻璃体描绘不同的子结构，最前部玻璃体是一个薄的胶原层，被称为前玻璃体。玻璃体核心包括整个中部地区的玻璃体。这部分的玻璃体是对比，这是足够的粘性，要把握钳和牢固地附着在底层的睫状体和视网膜玻璃体基底部的水。涵盖玻璃体的核心是一个非常薄的的称为玻璃体皮质的胶原外壳。 <img src="/files/ftp_upload/2455/2455fig2.jpg" alt="图2" /> 图2。玻璃体基底部的解剖结构。玻璃体基底部是沿锯齿缘（白色箭头），这是政企分开，睫状体和视网膜的分界线位于玻璃体半透明的下层结构。玻璃体基底部前缘延伸至睫状体玻璃体（白线）。玻璃体基底部后缘延伸2 - 3毫米，后到锯齿缘（白色虚线）。切除玻璃体基底部，镊子是用来把握组织和拉动了从底层的睫状体和视网膜。一旦升高，韦斯科特剪刀用于切割沿基地。 <img src="/files/ftp_upload/2455/2455fig3.jpg" alt="图3" /> 图3。一维玻璃体元素SDS - PAGE前玻璃体总蛋白的浓度，玻璃体基底部，维生素reous核心，玻璃体皮质分别为11.24，20.1，16.61，14.24毫克/毫升，分别。在200千伏为45分钟，凝胶电泳，染色与火烈鸟（BIO - RAD），和可视化使用VersaDoc成像系统（BIO - RAD）。为不同组织的剖面显示几个类似的乐队，说明无论是保守或交叉污染的蛋白质，以及独特的频带（星号），表明不同的本地化的蛋白质。

<ol>
	<li>Bishop, P. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+macromolecules+and+supramolecular+organisation+of+the+vitreous+gel.">Structural macromolecules and supramolecular organisation of the vitreous gel.</a> Prog Retin Eye Res. 19, 323-344 (2000).</li><li>Sebag, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+biology+of+pharmacologic+vitreolysis.">Molecular biology of pharmacologic vitreolysis.</a> Trans Am Ophthalmol Soc. 103, 473-494 (2005).</li><li>Yoshimura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comprehensive+analysis+of+inflammatory+immune+mediators+in+vitreoretinal+diseases.">Comprehensive analysis of inflammatory immune mediators in vitreoretinal diseases.</a> PLoS One. 4, e8158-e8158 (2009).</li><li>Gao, B. B., Chen, X., Timothy, N., Aiello, L. P., Feener, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+vitreous+proteome+in+diabetes+without+diabetic+retinopathy+and+diabetes+with+proliferative+diabetic+retinopathy.">Characterization of the vitreous proteome in diabetes without diabetic retinopathy and diabetes with proliferative diabetic retinopathy.</a> J Proteome Res. 7, 2516-2525 (2008).</li><li>Izuta, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+SOD+and+VEGF+are+increased+in+vitreous+bodies+from+proliferative+diabetic+retinopathy+patients.">Extracellular SOD and VEGF are increased in vitreous bodies from proliferative diabetic retinopathy patients.</a> Mol Vis. 15, 2663-2672 (2009).</li><li>Azimzadeh, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formalin-fixed+paraffin-embedded+(FFPE)+proteome+analysis+using+gel-free+and+gel-based+proteomics.">Formalin-fixed paraffin-embedded (FFPE) proteome analysis using gel-free and gel-based proteomics.</a> J Proteome Res. 9, 4710-4720 (2010).</li></ol>

玻璃体是一个半透明的凝胶，其分子组成是知之甚少，特别是在其子结构水平，玻璃体基底部的核心，皮质和前玻璃体。玻璃体核心包含胶原第二，第五，第九和第十一，，随着硫酸软骨素蛋白多糖，乙酰肝素硫酸蛋白多糖和透明质酸1,2蛋白质生物标志物在玻璃体核心疾病，如糖 ​​尿病性视网膜病变相关。3-5如何在每个子结构，这些蛋白质差异表达的特定蛋白质的身份，在很多情况下是不知道。这些细节可能会了解与特定的玻璃体视网膜疾病相关的蛋白质的来源，并帮助目标未来疗法。虽然尚未确定最佳组织解剖验尸间隔，蛋白质的降解可能影响下游实验。例如，免疫组织化学是在12小时死后眼睛的影响，一些特定的酶的活动可能会在几个小时内减少（未发表观察）。在这项研究中所有的组织，收集死亡的2至8小时，没有发生重大变化蛋​​白表达或适合蛋白质组学analsyis。液氮冷冻法保存超过固定选择，以防止蛋白质结构中的小变化引起的固定液的交联，已在其他组织中表现出的LC-MS/MS。蛋白质组学研究将取决于准确的能力剖析玻璃体不同的车厢，在这个视频的实验表明。我们已经证实的解剖技术，使用一维的SDS - PAGE。由于我们的研究结果表明，玻璃体中的各种子结构有差异表达的蛋白质。确定这些蛋白质会提供更详细地了解了玻璃体条块分割。

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 产品编号 </td></tr><tr><td> 0.12钳</td><td> STORZ Ophthalmics </td><td> E1502 </td></tr><tr><td> 5 - 40cc的注射器</td><td>流式细胞 - 迪金森</td><td> 309603 </td></tr><tr><td>随着锯齿直敷料镊</td><td> STORZ Ophthalmics </td><td> E1400 </td></tr><tr><td> 23号针</td><td>流式细胞 - 迪金森</td><td> 305145 </td></tr><tr><td>科利柏钳</td><td> STORZ Ophthalmics </td><td> 2 / 132 </td></tr><tr><td> Castroviejo角度角膜剪刀</td><td> STORZ Ophthalmics </td><td> E3223 </td></tr><tr><td> Vannas弧形截囊剪刀</td><td> STORZ Ophthalmics </td><td> E3387 </td></tr><tr><td> Weck - CEL手术矛</td><td>美敦力公司</td><td> 0008680 </td></tr><tr><td>韦斯科特弧形切断术剪刀</td><td> STORZ Ophthalmics </td><td> E3320 </td></tr></tbody></table>

dissection of human vitreous body elements for proteomic analysis

玻璃体是一个透明的，眼睛和覆盖在视网膜内的胶原细胞外基质，填充，1,2之间的玻璃体子结构和视网膜的异常相互作用的基础玻璃体视网膜疾病，包括视网膜裂孔和脱离，黄斑皱褶，黄斑裂孔，年龄相关性黄斑变性，玻璃体黄斑牵引，增生性玻璃体视网膜病变，增殖性糖尿病视网膜病变，并继承vitreoretinopathies 1,2玻璃体子结构的分子组成，是不知道。由于玻璃体是与有限的手术访问是透明的，它已经很难在分子水平上研究其子结构。我们开发了一种方法来分离和保存这些组织，蛋白质组学和生化分析。清扫技术在本实验视频演示如何隔离玻璃体基底部，前玻璃体，玻璃体的核心，从死后人类眼睛的玻璃体皮质。一维的每个玻璃体组件的SDS - PAGE分析表明，我们的清扫技术在四个独特的蛋白质图谱，对应于每个人的玻璃体下部结构。鉴定差异条块分割的蛋白质，就会发现潜在的各种玻璃体视网膜疾病的候选分子。

Watch this Scientific Journal Video about Dissection of Human Vitreous Body Elements for Proteomic Analysis at JoVE.com

Dissection of Human Vitreous Body Elements for Proteomic Analysis

该视频展示了一个区分和剖析各种半透明的人在验尸的眼睛玻璃体结构的有效技术。

人类蛋白质组分析玻璃体元素剖析

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

dissection-of-human-vitreous-body-elements-for-proteomic-analysis

Research

JoVE Journal

Medicine

31.7K 次观看.  University of Iowa. 该视频展示了一个区分和剖析各种半透明的人在验尸的眼睛玻璃体结构的有效技术。

视频: Dissection of Human Vitreous Body Elements for Proteomic Analysis

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.

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.

全玻璃体液清扫Vitreodynamic分析

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

科研

医学

Laser-capture Microdissection of Human Prostatic Epithelium for RNA Analysis

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types. Healthy prostate is comprised of fibromuscular stroma surrounding discrete epithelial-lined secretory lumens and a very small population of immune and neuroendocrine cells. In contrast, areas of prostate cancer have increased dysplastic luminal epithelium with greatly reduced or absent stromal population. Given the profound difference between stromal and epithelial cell types, it is imperative to separate the cell types for any type of downstream molecular analysis. Despite this knowledge, the bulk of gene expression studies compare benign prostate to cancer without micro-dissection, leading to stromal bias in the benign samples. Laser-capture micro-dissection (LCM) is an effective method to physically separate different cell types from a specimen section. The goal of this protocol is to show that RNA can be successfully isolated from LCM-collected human prostatic epithelium and used for downstream gene expression studies such as RT-qPCR and RNAseq.

The goal of this protocol is to use laser-capture micro-dissection as an effective method to isolate pure populations of cell types from heterogeneous prostate tissues for downstream RNA analysis.

人类前列腺上皮的激光捕获显微切割的RNA分析

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types.  Healthy prostate is comprised of ...

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.

解剖的小鼠胰腺的组织学分析与代谢谱

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.

小鼠尿的解剖和外植体培养对囊附着的作用分析

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

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

高脂喂养小鼠体内成分及代谢锁定分析

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

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 诱发主动脉瘤和解剖小鼠模型 Aortopathy 的定量显微 CT 分析

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

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

18F-fdg Petct 对活性人棕色脂肪组织的全身和区域定量

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Segmentation and Linear Measurement for Body Composition Analysis using Slice-O-Matic and Horos

Body composition is associated with risk of disease progression and treatment complications in a variety of conditions. Therefore, quantification of skeletal muscle mass and adipose tissues on Computed Tomography (CT) and/or Magnetic Resonance Imaging (MRI) may inform surgery risk evaluation and disease prognosis. This article describes two quantification methods originally described by Mourtzakis et al. and Avrutin et al.: tissue segmentation and linear measurement of skeletal muscle. Patients' cross-sectional image at the midpoint of the third lumbar vertebra was obtained for both measurements. For segmentation, the images were imported into Slice-O-Matic and colored for skeletal muscle, intramuscular adipose tissue, visceral adipose tissue, and subcutaneous adipose tissue. Then, surface areas of each tissue type were calculated using the tag surface area function. For linear measurements, the height and width of bilateral psoas and paraspinal muscles at the level of the third lumbar vertebra are measured and the calculation using these four values yield the estimated skeletal muscle mass. Segmentation analysis provides quantitative, comprehensive information about the patients' body composition, which can then be correlated with disease progression. However, the process is more time-consuming and requires specialized training. Linear measurements are an efficient and clinic-friendly tool for quick preoperative evaluation. However, linear measurements do not provide information on adipose tissue composition. Nonetheless, these methods have wide applications in a variety of diseases to predict surgical outcomes, risk of disease progression and inform treatment options for patients.

Segmentation and linear measurements quantify skeletal muscle mass and adipose tissues using Computed Tomography and/or Magnetic Resonance Imaging images. Here, we outline the use of Slice-O-Matic software and Horos image viewer for rapid and accurate analysis of body composition. These methods can provide important information for prognosis and risk stratification.

使用切片-O-马蒂奇和霍罗斯进行身体成分分析的细分和线性测量

Body composition is associated with risk of disease progression and treatment complications in a variety of conditions. Therefore, quantification of ...

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.

穆林·斯特拉特·甘利翁的位置、解剖和分析

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.

一种生成周期负荷诱导的大鼠膝关节内软骨病变模型的非侵入性方法

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