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1。前眼部の解剖。 <ol><li>角膜は、最初の15 °のブレード（図1）を用いて前房に角膜輪部で切開することによって除去される。その後、湾曲した角膜強膜はさみの一方のブレードは、前房に挿入されます。円周カットは、角膜輪部にちょうど前方、角膜に作られています。 </li><li> 0.12コリブリ鉗子とウェストコットの鋏は、毛様体の円周方向に前方アイリスを切り取るために使用されています。 </li><li>レンズ核は0.12コリブリ鉗子または15 °刃（supersharp）とピンセットを用い、皮質およびカプセルで削除されます。 </li></ol> 2。硝子体コア吸引。 <ol><li> 5 ccシリンジで23ゲージの針が挿入されます半ば硝子体（図1）。 </li><li>約硝子体コアを1 mL以上を静かに吸引される。 </li><li>次に、試料を遠心管にし、液体窒素中に配置されます。 </li></ol> 3。前部硝子体の解剖。 <ol><li>前部硝子体は、ガチョウの首の顕微鏡の光源からの入射光を調整することによって、半透明のリングとして見られている。硝子体コアの前の吸引は、簡単に識別するために他の構造体から前方の硝子体を分離する。 </li><li>弾性組織のシートは慎重にコリブリまたは麦粒鉗子と毛様体から離れて引っ張られ、Vannasのはさみでカットされます。この操作が繰り返されます。 </li><li>次に、試料を遠心管にし、液体窒素中に配置されます。 </li></ol> 4。硝子体ベースの解剖。 <ol><li>アイカップを安定させるために0.12鉗子を使用して、ウェストコットのはさみは、視神経乳頭に向かって毛様体からの4つの等間隔のストレス解消になるの削減を行うことによって&quot;花&quot;の目に使用されます。 </li><li>毛様体（図2）の同価のplicataはウェストコットのはさみを使用して、象限のそれぞれから削除されます。 </li><li>硝子体のベースは、毛様扁平と網膜（図2）上の鉗子と鋸状縁のどちら側でも把握している。 </li><li>硝子体ベース上で連続的な牽引力は鉗子で適用されます。ウェストコットのはさみを持つシーケンシャル切削は、それを摘出されている外観&quot;真珠の数珠&quot;と半透明の組織を抽出する。 </li><li>次に、試料を遠心管にし、液体窒素中に配置されます。 </li></ol> 5。硝子体皮質除去。 <ol><li>パルスプラナは鋸状縁（図2）の後、約3 mmのリーフレットをカットしてウェストコットのはさみと象限の各々から切り出される。 </li><li>組織のリーフレットとの間で、硝子体皮質が、半透明フィルムとして可視化される。 </li><li>弾性膜はどちら0.12鉗子で把持またはベックリキセル手術用スポンジへの遵守が保有しています。硝子体皮質は、その後離れて網膜から引っ張られ、ウェストコットハサミ（図1）で切り出しています。 </li><li>サンプルは、マイクロ遠心チューブにして、液体窒素内に配置されます。実験のために利用されるまで、すべてのサンプルは-80℃で保存されています。 </li></ol> 6。代表的な結果組織サンプルは、特定の実験のための様々な方法で処理することができます。私たちのケースでは、サンプルはSDS - PAGE（図3）によってタンパク質分析のために提出された。 <img src="/files/ftp_upload/2455/2455fig1.jpg" alt="図1" /> 図1。硝子体の別の部分構造を描いた人間の眼の断面図。最前硝子体は薄いコラーゲン層前部硝子体と呼ばれる。硝子体コアは、硝子体の全体の中央領域を含む。硝子体のこの部分には、鉗子で把持するのに十分な粘性であり、しっかりと基礎と毛様体と網膜に接続されているガラスベースとは対照的に、より多くの水です。硝子体コアを包含する硝子体皮質と呼ばれる非常に薄いコラーゲンのシェルです。 <img src="/files/ftp_upload/2455/2455fig2.jpg" alt="図2" /> 図2。硝子体の基本解剖学は。硝子体基部には毛様体や網膜を分離する境界線である鋸状縁（白矢印）、に沿って位置硝子体の半透明の部分構造である。硝子体基部の前縁には毛様体の毛様扁平（白い線）にわたって広がっている。硝子体基部の後縁は、ORAのコナラ（白ダッシュ）の後2〜3 mmを拡張します。消費税​​に硝子体基部には、鉗子は、組織を把握し、基盤となる毛様体および網膜からそれを引き離すために使用されます。一度上昇、ウェストコットはさみ、ベースに沿って切断するために使用されます。 <img src="/files/ftp_upload/2455/2455fig3.jpg" alt="図3" /> 図3。硝子体の要素の一次元SDS - PAGE。前部硝子体、硝子体ベース、VITの総タンパク質濃度reousコア、および硝子体皮質はそれぞれ11.24、20.1、16.61、14.24 mg / mLのであった。ゲル電気泳動は、45分間、200kVで行ったフラミンゴ（Bio - Rad社）で染色し、そしてVersaDocイメージングシステム（Bio - Rad）を用いて可視化した。異なる組織のプロファイルは、差動ローカライズされた蛋白質を示し、（アスタリスク）保存やタンパク質の相互汚染だけでなく、ユニークなバンドのいずれかを示す、いくつかの同様のバンドを示す。

<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>

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硝子体ベース、コア、皮質、および前部硝子体：硝子体は、その分子組成の悪い、特にその部分構造のレベルで、理解されている半透明のジェルです。硝子体コアは、糖尿病性網膜症などの疾患に関連付けられているコンドロイチン硫酸プロテオグリカン、ヘパラン硫酸プロテオグリカン、および硝子体コアのヒアルロン酸。1,2タンパク質のバイオマーカーと一緒に、コラーゲンII、V、IX、およびXIが含まれています。3-5方法これらのタンパク質は、差動部分構造のそれぞれで表され、そして多くの場合、特定のタンパク質の同一性は、知られていない。これらの詳細については、特定の網膜硝子体疾患に関連するタンパク質の起源への洞察を与え、ターゲットの将来の治療を助けるかもしれない。組織郭清のための最適な死後の間隔が決定されていないが、タンパク質分解は、下流の実験に影響を与える可能性があります。例えば、免疫組織化学は、12時間の死後の目といくつかの特定の酵素の活動に影響を受けています（未発表観察）、数時間以内に短縮することができます。本研究ではすべての組織は、タンパク質の発現やプロテオミクスのanalsyisへの適合性を大幅に変更することなく、死の2と8時間の間に採取された。保全の液体窒素凍結法は、LC-MS/MSによって他の組織で実証されているリンクを固定クロス、によって引き起こされる蛋白質の構造の小さな変化を防ぐために、固定優先して選択される6プロテオミクス研究は、正確に能力に依存しますとしてこのビデオの実験で示されて硝子体の異なる区画を、詳細に分析します。我々は1次元SDS - PAGEを用いて解剖手法を検証しています。我々の結果が示唆するように、差動、様々な硝子体のサブ構造体中のタンパク質をそこに表現されています。これらのタンパク質を同定することは硝子体のコンパートメントのより詳細な理解を提供します。

<table border="1"><tbody><tr><td> の名前 </td><td> 会社 </td><td> カタログ番号 </td></tr><tr><td> 0.12鉗子</td><td>シュトルツ眼科</td><td> E1502 </td></tr><tr><td> 5 ccシリンジ</td><td>ベクトンディッキンソン- </td><td> 309603 </td></tr><tr><td>セレーション付きストレート麦粒鉗子</td><td>シュトルツ眼科</td><td> E1400 </td></tr><tr><td> 23ゲージの針</td><td>ベクトンディッキンソン- </td><td> 305145 </td></tr><tr><td>コリブリ鑷子</td><td>シュトルツ眼科</td><td> 132分の2 </td></tr><tr><td> Castroviejo角度角膜ハサミ</td><td>シュトルツ眼科</td><td> E3223 </td></tr><tr><td> Vannas曲線嚢切開のはさみ</td><td>シュトルツ眼科</td><td> E3387 </td></tr><tr><td>ベックリキセル外科スピアーズ</td><td>メドトロニック</td><td> 0008680 </td></tr><tr><td>ウェストコット曲線腱切除のはさみ</td><td>シュトルツ眼科</td><td> E3320 </td></tr></tbody></table>

dissection of human vitreous body elements for proteomic analysis

硝子体は眼球と覆って網膜の内側を満たす光学的に透明な、コラーゲン細胞外マトリックスである。硝子体下部構造と網膜の間に1,2異常の相互作用は網膜裂傷や剥離、黄斑パッカ、黄斑円孔を含むいくつかの網膜硝子体疾患を、根底加齢黄斑変性症、vitreomacularのトラクション、増殖性硝子体網膜症、増殖性糖尿病網膜症、および継承されたvitreoretinopathies。1,2硝子部分構造の分子組成が知られていない。硝子体は、限定された外科的アクセスが透過的に行われるので、それは分子レベルでその下部構造を勉強することは困難であった。我々は、プロテオミクスと生化学的解析のために、これらの組織を分離して保存する方法を開発した。この実験的な映像で解剖手法は、死後、人間の眼から硝子体基部、前部硝子体、硝子体コア、および硝子体皮質を分離する方法を示しています。各ガラス成分の一次元SDS - PAGE分析では、私たちの解剖技術が人間の硝子体の各部分構造に対応する4つのユニークな蛋白質のプロファイルをもたらしたことを示した。差動区画化タンパク質の同定は、様々な網膜硝子体疾患の根底にある候補分子を明らかにする。

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

このビデオでは、死後の目には人間の硝子体の様々な半透明の構造を差別化し、解剖のための効果的なテクニックを示しています。

プロテオーム解析のための人間の硝子体の要素の解剖

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

Nanyang Technological University

Research

JoVE Journal

Medicine

31.8K ビュー.  University of Iowa. このビデオでは、死後の目には人間の硝子体の様々な半透明の構造を差別化し、解剖のための効果的なテクニックを示しています。

ビデオ: Dissection of Human Vitreous Body Elements for Proteomic Analysis

Network Analysis of the Default Mode Network Using Functional Connectivity MRI in Temporal Lobe Epilepsy

Functional connectivity MRI (fcMRI) is an fMRI method that examines the connectivity of different brain areas based on the correlation of BOLD signal fluctuations over time. Temporal Lobe Epilepsy (TLE) is the most common type of adult epilepsy and involves multiple brain networks. The default mode network (DMN) is involved in conscious, resting state cognition and is thought to be affected in TLE where seizures cause impairment of consciousness. The DMN in epilepsy was examined using seed based fcMRI. The anterior and posterior hubs of the DMN were used as seeds in this analysis. The results show a disconnection between the anterior and posterior hubs of the DMN in TLE during the basal state. In addition, increased DMN connectivity to other brain regions in left TLE along with decreased connectivity in right TLE is revealed. The analysis demonstrates how seed-based fcMRI can be used to probe cerebral networks in brain disorders such as TLE.

The Default Mode Network (DMN) in Temporal Lobe Epilepsy (TLE) is analyzed in the resting state of the brain using seed-based functional connectivity MRI (fcMRI).

側頭葉てんかんにおける機能的結合MRIを用いてデフォルトモードネットワークのネットワーク解析

Functional connectivity MRI (fcMRI) is an fMRI method that examines the connectivity of different brain areas based on the correlation of BOLD signal ...

医学

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.

Vitreodynamic分析のための全硝子体液解剖

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

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.

RNA分析のためのヒト前立腺上皮のレーザーキャプチャーマイクロダイセクション

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.

マウスの膵臓の組織学的解析と代謝プロファイリングに対する郭清

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.

郭清と尿の添付ファイルのIn Vitro解析マウス尿膜の培養

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.

身体組成と高脂肪給電マウスにおける代謝ケージ分析

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.

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.

Aortopathy β-アミノプロピオニ トリルによる大動脈瘤と解離のマウスモデルでのマイクロ CT 定量分析

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

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.

スライスO-マティックとホロを用いた体組成解析のためのセグメンテーションと線形測定

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.

ネズミ星状神経節の位置、解剖、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.

ラット膝の周期的負荷誘発関節内軟骨病変モデルを生成するための非侵襲的方法

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