著者は、技術的な相談のための博士アナCotrimへと機器のMurshed博士Monzurに感謝しています。この作品は、部分的に保健研究のためのカナダの協会によって資金を供給された。

1。手続き的説明<ol><li>皮下輸液の動きを中断することから唾液の分泌を防ぐために、各マウスには0.5 mg / mlの硫酸アトロピン一水和物（カタログ番号11330、フルカ、米国）の1g /μlの体重を注入する。 </li><li> 100 mg / mLのケタミンおよび20 mg / mlのキシラジン（フェニックスサイエンティフィック、AR、米国）ソリューションの1g /μlの体重で各マウスに麻酔注入し腹腔内 （IP） </li><li>腹側の位置にカスタムメイドのプラスチックのプラットフォーム上でマウスを置きます：上顎切歯は、金属線にロックされ、下顎切歯は、口が開いて保持するために弾性文字列にフックされます。 </li><li> 0.58ミリメートルの直径のポリエチレンチューブ（; 図2猫＃427410、Becton Dickinson社製、MD、米国）内に配置された29ゲージの針（カタログ番号600145、タイコヘルスケア、MA、米国）とインスリンの注射器を使用してください。ポリエチレン管は、29ゲージの針の先端上に5ミリメートル挿入されます。針は、ワルトン管にポリエチレンチューブを移動するため硬質のガイドとして使用されます。ポリエチレン管は、ダクト内部に3〜5ミリメ​​ートルを挿入されます。針はダクトの外側に残っている間だけのポリエチレン管は、唾液腺管に入ることに注意してください。右側の舌の下に置かれているワルトン管を介して顎下腺にフィルタ処理された0.4％トリパンブルー染色液（カタログ番号15250、ギブコ、米国）の50ULを注入する。注入は注射器のピストン上で正圧に着実に供給される指は、注入のすべての後に口腔内注入の逆流を防ぐために、腺に配信されていても、ピストン上に残ります。トリパンブルー溶液は、技術を実証するために染料としてビデオで使用されています。 </li><li> FTERトリパンブルー溶液の注入は、マウスを解剖する前に安楽死された。唾液腺を外科的にどのくらいの輸液のは、実際に腺に残っているかを判断するために公開されます。 </li></ol> 2。代表的な結果： 輸液を注入した後、マウスを外科的に後のカニューレ唾液腺を露出する解剖した。画像- 2に示すように、ワルトン管の暗色はトリパンブルー溶液が正常に注入されたという証拠を提供する。 <img src="/files/ftp_upload/3074/3074fig1.jpg" alt="図1" /> 図1マウスの顎下腺のカニューレ。 <img src="/files/ftp_upload/3074/3074fig2.jpg" alt="図2" /> 図2。ポリエチレン管は29ゲージの針の先端の周りに包み込まれている。注射針は、ワルトン管にポリエチレンチューブを移動するため硬質のガイドとして使用されます。ポリエチレン管は、ダクトの内部に（カニューレ）3-5 mmに挿入されます。針はダクトの外側に残っている間だけのポリエチレン管は、唾液腺管に入ることに注意してください。

<ol>
	<li>Fox, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+salivary+dysfunction.+Drugs+and+radiation.">Acquired salivary dysfunction. Drugs and radiation.</a> Ann N Y Acad Sci. 842, 132-137 (1998).</li><li>Bhide, S. A., Miah, A. B., Harrington, K. J., Newbold, K. L., Nutting, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation-induced+xerostomia:+pathophysiology,+prevention+and+treatment.">Radiation-induced xerostomia: pathophysiology, prevention and treatment.</a> Clin Oncol (R Coll Radiol). 21, 737-744 (2009).</li><li>Redman, R. S., Ball, W. D., Mezey, E., Key, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dispersed+donor+salivary+gland+cells+are+widely+distributed+in+the+recipient+gland+when+infused+up+the+ductal+tree.">Dispersed donor salivary gland cells are widely distributed in the recipient gland when infused up the ductal tree.</a> Biotech Histochem. 84, 253-260 (2009).</li></ol>

全ての実験は、大学の動物ケア委員会によって承認され、動物のケアに関するカナダの協議会のガイドラインに準拠していましたされています。

このカニューレ挿入法を使用して難しさは、注入溶液の逆流を防ぐためです。トリパンブルー溶液を一時的に正常唾液腺に移行した場合でも、唾液の分泌は、注入を妨害し、腺の外トリパンブルー液の逆流が発生する場合があります。予防策として、唾液の分泌を抑制するためにアトロピンの使用に加え、ポリエチレンカニューレは、ワルトン管や点滴が堆積された後、60秒間保持シリンジ?...

<table border="1"><tbody><tr><td> 試薬の名前 </td><td> タイプ </td><td> 会社 </td><td> カタログ番号 </td><td> コメント（省略可能） </td></tr><tr><td>アトロピン</td><td>薬</td><td>フルカ</td><td> 11330 </td><td>唾液の分泌を抑制する</td></tr><tr><td>ケタミン</td><td>薬</td><td>フェニックスサイエンティフィック</td><td></td><td>鎮静状態</td></tr><tr><td>キシラジン</td><td>薬</td><td>フェニックスサイエンティフィック</td><td></td><td></td></tr><tr><td>ポリエチレンチューブ（0.58 mm径） </td><td>チューブ</td><td> Intramedic PE - 10、ベクトンディッキンソン診断システム</td><td> 427410 </td><td>ワルトン管への挿入中に組織の損傷を避けるために、針の先端を介して挿入する。 </td></tr><tr><td>ウルトラコンフ​​ォートインスリン注射器（29G針付） </td><td>注射器</td><td>タイコヘルスケア</td><td> 600145 </td><td></td></tr><tr><td> 4パーセントトリパンブルー</td><td>染色</td><td>ギブコ</td><td> 15250 </td><td></td></tr></tbody></table>

cannulation of the mouse submandibular salivary gland via the wharton's duct

重度の唾液腺の機能低下が頻繁にシェーグレン症候群と癌治療のために彼らの頭頸部領域での治療照射を受けている人の患者に見られる。このような口腔乾燥（口渇）、嚥下障害（咀嚼や嚥下障害）などの患者の経験の症状の両方のグループ、重度の虫歯、味覚変化、オーロ-咽頭感染症（カンジダ症）、粘膜炎、痛みや不快感。 唾液腺の低刺激機能の治療のための再生医療の一つの革新的なアプローチはRSレッドマン、E Mezey ら推測される2009：。幹細胞を直接障害の機能を復活さで強力な方法として、グランドにカニュレーションによって堆積することができるオルガン。おそらく、移行した外国の幹細胞は、宿主唾液腺の一部として機能する腺細胞に分化する。また、このカニューレ挿入法は、臨床的遺伝子導入のアプリケーションのための方便と効果的な配信方法です。 ここでは、ワルトン管（ 図1）を経由してマウスの顎下唾液腺にカニュレーションの手順を実行するために必要な手順を示しています。 C3Hマウス（チャールズリバー、モントリオール、QC、カナダ）は、マギル大学動物資源センターでクリーンな、従来の条件下で保持されているこの実験に使用されます。全ての実験は、大学の動物ケア委員会によって承認され、動物のケアに関するカナダの協議会のガイドラインに準拠していましたされています。 この実験では、トリパンブルー溶液を29ゲージの針ポリエチレン管の内側に包まれてインスリンの注射器を使用してワルトン管の開口部を介してグランドに注入する。その後、マウスは注入が正常腺に移行することを示すために解剖される。

Watch this Scientific Journal Video about Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct at JoVE.com

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton&#039;s Duct

ワルトン管を介してマウス顎下唾液腺のカニュレーションのためのプロトコルが記述されています。この実験では、トリパンブルー溶液は、この手法が効果的に標的腺に注入を提供する方法を示すこと、および唾液腺の再生のための潜在的な臨床薬/細胞療法としてのこの新しいアプローチの信頼性を示唆するダイアーとして使用されます。

ワルトン管を経由してマウス顎下唾液腺のカニュレーション

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

cannulation-mouse-submandibular-salivary-gland-via-wharton-s

Research

JoVE Journal

Medicine

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

25.7K ビュー.  McGill University. ワルトン管を介してマウス顎下唾液腺のカニュレーションのためのプロトコルが記述されています。この実験では、トリパンブルー溶液は、この手法が効果的に標的腺に注入を提供する方法を示すこと、および唾液腺の再生のための潜在的な臨床薬/細胞療法としてのこの新しいアプローチの信頼性を示唆するダイアーとして使用されます。

ビデオ: Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

唾液を収集し、家族介護者を経由して高齢者を居住する虚弱コミュニティ唾液コルチゾールおよびα-アミラーゼの測定

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

医学

Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide new expandable sources of beta cells to alleviate the donor scarcity in human islet transplantation as therapy for diabetes. Although recent advances have been made towards this aim, mechanisms that regulate beta cell expansion and differentiation from a stem/progenitor cell remain to be characterized. Here, we describe a protocol for an injury model in the adult mouse pancreas that can function as a tool to study mechanisms of tissue remodeling and beta cell proliferation and differentiation. Partial duct ligation (PDL) is an experimentally induced injury of the rodent pancreas involving surgical ligation of the main pancreatic duct resulting in an obstruction of drainage of exocrine products out of the tail region of the pancreas. The inflicted damage induces acinar atrophy, immune cell infiltration and severe tissue remodeling. We have previously reported the activation of Neurogenin (Ngn) 3 expressing endogenous progenitor-like cells and an increase in beta cell proliferation after PDL. Therefore, PDL provides a basis to study signals involved in beta cell dynamics and the properties of an endocrine progenitor in adult pancreas. Since, it still remains largely unclear, which factors and pathways contribute to beta cell neogenesis and proliferation in PDL, a standardized protocol for PDL will allow for comparison across laboratories.

This protocol describes an injury model involving the surgical ligation of the pancreatic duct in the adult mouse pancreas, resulting in severe injury that establishes an environment that allows beta cell neogenesis and proliferation. This model can be used as a tool to study mechanisms involved in beta cell formation.

内分泌と外分泌リプログラミングおよび増殖のためのモデルとしての膵管の連結を介して、マウスの膵臓への外科的損傷

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. Irreparable cell damage results in a chronic loss of salivary function that impairs basic oral activities, and increases the risk of oral infections and dental caries. Salivary hypofunction and its complications gravely impact a patient's comfort. Current symptomatic management of the condition is ineffective, and newer therapies to assuage the condition are needed. 
Salivary glands are exocrine glands, which expel their secretions into the mouth via excretory ducts. Cannulation of these ducts provides direct access to the glands. Retroductal delivery of a contrast agent to major salivary glands is a routine out-patient procedure for diagnostic imaging. Using a similar procedure, localized treatment of the glands is feasible. However, performing this technique in preclinical studies with small animals poses unique challenges. In this study we describe the technique of retroductal administration in rat submandibular glands, a procedure that was refined in Dr. Bruce Baum's laboratory (NIH)1, and lay out a procedure for local gland irradiation.

Salivary gland hypofunction, a major adverse effect of head and neck radiotherapy diminishes a patient's quality of life. The demonstration of efficacy of new therapies in animal models is a prerequisite before clinical transition. This protocol describes retroductal administration and local irradiation of rat submandibular glands.

唾液機能低下のラットモデルにおけるRetroductal顎下腺点滴とローカライズされた分割照射

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. ...

Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology. There are few tools to further these areas of study. Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. The technique requires the intricate dissection of the second portion of the duodenum and ampulla, followed by the occlusion of the bile duct and the cannulation of the pancreatic duct. Although the technique is technically challenging at first, the applications are myriad. Ambiguity in the specifics of the procedure between groups highlighted the need for a standard protocol. This work describes the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery. The infusion and therefore expression is specific to the pancreas, without expression present in any other tissue type.

Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. A pancreatic duct infusion technique for drug and viral delivery to pancreatic cells is presented here.

膵管注入:薬物およびウイルス送達の有効かつ選択的な方法

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, ...

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In these cases, questions remain regarding disease pathogenesis and effective interventions. An optimized technique that allows functional assessment of the salivary glands is invaluable for investigating exocrine gland biology, dysfunction, and therapeutics. Here, we present a step by step approach to performing pilocarpine stimulated saliva secretion, including tracheostomy and the dissection of the three major murine salivary glands. We also detail the appropriate murine head and neck anatomy accessed during these techniques. This approach is scalable, allowing for multiple mice to be processed simultaneously, thus improving the efficiency of the work flow. We aim to improve the reproducibility of these methods, each of which has further applications within the field. In addition to saliva collection, we discuss metrics for quantifying and normalizing functional capacity of these tissues. Representative data are included from submandibular glands with depressed salivary gland function 2 weeks following fractionated radiation (4 doses of 6.85 Gy).

We present a detailed approach to performing saliva collection, including murine tracheostomy and the isolation of three major salivary glands.

ピロカルピン刺激次を介してマウス唾液機能的アセスメント分別放射線

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In ...

Radiation Treatment of Organotypic Cultures from Submandibular and Parotid Salivary Glands Models Key In Vivo Characteristics

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with radiotherapy. Experimental approaches to understanding mechanisms of salivary gland dysfunction and restoration have focused on in vivo models, which are handicapped by an inability to systematically screen therapeutic candidates and efficiencies in transfection capability to manipulate specific genes. The purpose of this salivary gland organotypic culture protocol is to evaluate maximal time of culture viability and characterize cellular changes following ex vivo radiation treatment. We utilized immunofluorescent staining and confocal microscopy to determine when specific cell populations and markers are present during a 30-day culture period. In addition, cellular markers previously reported in in vivo radiation models are evaluated in cultures that are irradiated ex vivo. Moving forward, this method is an attractive platform for rapid ex vivo assessment of murine and human salivary gland tissue responses to therapeutic agents that improve salivary function.

Using three-dimensional organotypic cultures to visualize morphology and functional markers of salivary glands may provide novel insights into the mechanisms of tissue damage following radiation. Described here is a protocol to section, culture, irradiate, stain, and image 50&#8211;90 &#956;m thick salivary gland sections prior to and following exposure to ionizing radiation.

顎下および耳下腺唾液腺からの Organotypic 培養の放射線治療は生体内特性をモデル化する

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

胆管TNBS輸液による慢性膵炎のマウスモデル

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...

Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a lack of effective treatments in the current clinical practice. With the easy accessibility of transgenic and knockout strains and their small size, which allows minimal doses of drugs required for in vivo evaluation, a well-established experimental model in mice is preferred for AP research. Moreover, SAP induced through sodium taurocholate (TC) is currently one of the most widely used and best characterized models. This model has been investigated for novel therapies and possible molecular events during the process of AP. Here, we present the generation of an AP mouse model using sodium taurocholate and a simple homemade microsyringe. Moreover, we also provide the methodology for the subsequent histology and serological testing.

A mouse model of severe acute pancreatitis is described herein. The procedure presented here is very rapid, simple, and accessible, thereby potentially allowing the study of the molecular mechanisms and different therapeutic interventions in acute pancreatitis in a convenient way.

ビリオ膵管へのタウロコール酸ナトリウムの逆行性注入を用いたマウス重症急性膵炎モデルの確立

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a ...

Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is similar in color to the thyroid gland. The lack of effective animal models in preclinical research is a severe limitation for the development of PG identification techniques. This protocol allows for the establishment of a simple and effective rat model for PG identification. In this model, black iron oxide nanoparticles (IONPs) are injected locally in the thyroid gland and rapidly diffuse within the thyroid gland but not the PG. A negatively stained PG and a positively stained thyroid gland can be easily identified by the naked eye without requiring external microscopes. The position of the PG can be identified by increasing the color contrast between the thyroid gland and the PG, based on the color of the black IONPs. This rat model is low-cost and convenient for PG identification, and the IONPs are a novel PG contrast agent.

To date, the development of parathyroid gland (PG) identification methods is limited by the lack of animal models in preclinical research. Here, we establish a simple and effective rat model for intraoperative PG imaging and evaluate its effectiveness by using iron oxide nanoparticles as a novel PG contrast agent.

術中副甲状腺イメージングのための簡便かつ効果的なラットモデルの確立

Parathyroid gland (PG) identification is a critical unmet need in thyroidectomy. The identification of the PG is challenging in thyroid surgery as it is ...

Laparoscopic Common Bile Duct Exploration in Patients with a Previous History of Biliary Tract Surgery

For recurrent choledocholithiasis, abdominal adhesions in previous surgeries lead to changes in anatomical structures, and a secondary injury occurs easily when performing another operation for laparoscopic common bile duct exploration (LCBDE), which was once considered a relative contraindication. In view of the limitations of the current surgical technique, this study summarized the surgical approaches and crucial anatomical landmarks for reoperation for LCBDE. Four general surgical approaches were proposed to expose the common bile duct, including the ligamentum teres hepatis approach, the anterior hepatic duodenal ligament approach, the right hepatic duodenal ligament approach, and the hybrid approach. Additionally, this study highlighted seven crucial anatomical landmarks: the parietal peritoneum, the gastrointestinal serosa, the ligamentum teres hepatis, the inferior margin of the liver, the gastric antrum, the duodenum, and the hepatic flexure of the colon, which were helpful to safely separate abdominal adhesions and expose the common bile duct. Moreover, to shorten the time of choledocholithotomy, a sequential method was innovatively applied for the removal of the stones in common bile duct. Mastering the above surgical approaches, including identifying crucial anatomical landmarks and adopting the sequential method will improve the safety of reoperation for LCBDE, shorten the operation time, promote the fast recovery of patients, reduce postoperative complications, and contribute to the popularization and application of this technique.

Programmed surgery based on surgical methods and anatomical markers helps shorten the operation time, reduce complications, and improve the safety of the surgery. This study investigated and summarized the surgical methods and anatomic markers of laparoscopic common bile duct exploration in patients with a history of biliary tract surgery.

胆道手術の既往歴のある患者における腹腔鏡下総胆管探索

For recurrent choledocholithiasis, abdominal adhesions in previous surgeries lead to changes in anatomical structures, and a secondary injury occurs ...