根尖切除小鼠模型来研究早期哺乳动物心脏再生
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
心脑血管疾病困扰着整个世界，由于密集的生活方式的改变。心脏再生持有修复和恢复因受伤和疾病失去了心肌巨大潜力。相反，某些低等脊椎动物的强劲心脏再生，成年哺乳动物心脏通常显示为心脏的再生和修复能力最低。然而，最近的研究已经引起了相当大的科学兴趣地注意到，产后天之间1〜7（P1到P7），新生小鼠心脏保留显著再生潜能根尖切除术（ 即 ，手术截肢和左心室心尖曝光）后的发现。超过这一发现一个主要的争论可能是由于不同的手术相关的程序中的努力来复制或扩大在此重要的发现使用。这些指令动态地呈现材料和方法根尖切除的小鼠模型。其显着的步骤该啮齿动物surviva的升手术涉及低温麻醉，开胸，心脏心室心尖手术截肢，缝合和老鼠的恢复。描述可以扩大根尖切除的小鼠模型的心血管研究的应用程序的方法。
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
作者感谢博士。詹姆斯·霍金斯，祖熙玉和禤去从国家心脏，肺和血液研究所（NHLBI）为他们的鼠标手术准备和石蜡切片染色的援助。作者感谢美国国立卫生研究院研究员编委会的编辑协助。
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 |
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