アピカル切除マウスモデルは、初期の哺乳類の心臓の再生を研究するために
Summary
A step-by-step video protocol of apical resection is demonstrated in this study. Apical resection is a recently highlighted surgical approach in mammalian heart regeneration research. This study may promote the application of apical resection as a standard methodology in research into the mechanism underlying heart regeneration.
Abstract
心血管疾患は集中的なライフスタイルの変化による全世界を悩ませています。ハートの再生には、負傷や病気に負けた心筋細胞を修復し、復元するための大きな可能性を秘めています。一定の下等脊椎動物の強固な心臓再生とは対照的に、大人の哺乳類の心臓は、通常、心臓の再生および修復のための最小限の容量を示しています。しかし、最近の研究では、（P7へP1）1〜7生後日の間、新生児マウスの心臓が心尖切除（ すなわち 、外科的切断および左心室の頂点の曝露）後の重要な再生能力を保持し、知見とかなりの科学的関心を巻き起こしています。この発見を介して1つの主要な論争は、この重要な知見に複製または拡大するための努力で使用される多様な手術に関連した手順に起因する可能性があります。これらの命令は、動的にマウスモデルにおいて、頂端切除するための材料および方法論を提示します。このげっ歯類survivaの顕著なステップL手術は低体温麻酔、開胸、心臓心室尖部の外科的切断、縫合糸およびマウスの回復を伴います。記載されているアプローチは、心血管研究のための頂端切除マウスモデルの適用を拡大することができます。
Introduction
Prolonged human life span leads to various aging- and lifestyle-related diseases, including heart failure, a leading cause of mortality. However, without replacement of lost or dysfunctional cardiac muscle cells, current therapeutics can only transiently improve cardiac function1, 2. Thus, it is necessary to discover and develop innovative strategies for cardiac regeneration and repair. The adult mammalian heart has limited regenerative potential. Studies from lower vertebrates, such as urodele amphibians and teleost fish, have provided unprecedented insights into the molecular and cellular mechanisms underlying heart regeneration3, 4. Recently, a neonatal mouse model of heart regeneration has emerged that might enable identification and characterization of more evolutionarily conserved pathophysiological events required for human heart regeneration5.
Apical resection refers to the surgical removal of left ventricular apex. This procedure is restricted to 1- to 7-day-old (P1 to P7) mice due to its high lethality in older mice6. The cardiac regeneration process in neonatal mice after apical resection is expected to be as follows: (I) rapid and effective formation of a hematoma to seal the apex and prevent exsanguination; (II) cardiomyocyte regeneration and restoration of systolic function5, 7. Recent work has stimulated debate on the significance and efficiency of this model8-11. Thus, it is important to present the apical resection clearly and in detail. To this end, this protocol vividly and specifically describes a video of how I did apical resection based on a previous protocol6.
Understanding the molecular mechanisms underlying cardiac regeneration is of importance for treatment of heart disease characterized by loss and/or injured cardiomyocytes, such as heart failure1, 2. Given the current and promising progress of apical resection in the research of cardiac regeneration, this study could promote the use of this technique and its uses in cardiac regeneration research.
Protocol
Representative Results
Discussion
Cardiac regeneration shows potential for the treatment and prevention of heart failure1, 2. Animal models are indispensable and play a critical role in understanding how cardiac regeneration occurs3-6. Many amphibians and fish regenerate heart tissue in response to injury, providing insight into our understanding of human cardiovascular disease13, 14. In terms of evolution, however, the pathophysiology should be more conserved between a mouse model and humans. Heart regeneration in mammal…
Declarações
The authors have nothing to disclose.
Acknowledgements
著者は、博士に感謝します。マウスの手術と準備し、パラフィン切片の染色との支援のための国立心肺血液研究所（NHLBI）からジェームズ・ホーキンス、祖-XIゆうと玄屈原。著者は、編集支援のためのNIHフェロー編集委員会に感謝しています。
Materials
| Olsen-Hegar Needle Holders with Scissors,1.5mm | Fine Science Tools | 12002-12 | |
| Vannas Spring Scissors – 2mm Cutting Edge | Fine Science Tools | 15000-03 | Iridectomy scissors |
| Hot Bead Sterilizer | Fine Science Tools | 18000-45 | |
| Iris Forceps, Straight, Serrated | Fine Science Tools | 11064-07 | |
| Iris Forceps, Curved, Serrated | Fine Science Tools | 11065-07 | |
| Shea Scissors – Curved/Blunt-Blunt/12cm | Fine Science Tools | 14105-12 | |
| Round Handled Suture Tying Forceps, Straight | Fine Science Tools | 18025-12 | |
| Round Handled Vannas Spring Scissors | Fine Science Tools | 15400-12 | |
| Magnifying Lamp | Luxolamp Corp | IM120 | |
| Heating lamp | Brandt Equipment llc | 51152/3 | |
| 6-0 Prolene sutures | Ethicon | 8889H | |
| Skin glue-Vetbond Tissue Adhesive | 3M | 1469 | |
| Sterile Cotton Tipped Applicators | Dynarex | 4305 | |
| WEBCOL Alcohol Prep Pad | Covidien | 6818 | Medium 2 PLY, 200/BOX, Satured with 70% Isopropyl Alcohol |
| Curity All Purpose Sponges | Covidien | 9024 | Non-woven 4 PLY, 4"x4" (10.2cm×10.2cm) |
| Bench top protector sheet | KIMTECH SCIENCE | 7546 | 18" x 19.5" (45.72cm x 49.53cm) x 50 |
| 0.9% Sodium Chloride, 250ml | Hospira Inc. | NDC 0409-6138-22 | |
| Betadine Solution Swabsticks | Purdue Products L.P. | NDC 67618-153-03 | |
| Autoclave | TOMY Digital Biology | SX-700 | HIGH-PRESSURE STEAM STERILIZER |
| Slide scanner | HAMAMATSU | NanoZoomer 2.0-RS |
Referências
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