A ressecção apical mouse modelo para estudar precoce Regeneração Mammalian Coração
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
A doença cardiovascular assola o mundo inteiro devido a mudanças de estilo de vida intensivos. A regeneração do coração é uma grande promessa para reparar e restaurar cardiomiócitos perdidos devido a lesões e doenças. Em contraste com a regeneração cardíaca robusta de certos vertebrados inferiores, corações de mamíferos adultos mostram tipicamente uma capacidade mínima para a regeneração cardíaca e reparação. No entanto, estudos recentes têm suscitado interesse científico considerável com a constatação de que, entre o dia pós-natal 1-7 (P1 a P7), o coração neonatal do rato mantém potencial regenerativo significativa após a ressecção apical (ou seja, a amputação cirúrgica e exposição do ápice ventricular esquerda). Uma grande controvérsia sobre este achado pode ser devido aos procedimentos relacionadas à cirurgia diversos usados em esforços para replicar ou expandir sobre esta importante descoberta. Estas instruções apresentar dinamicamente os materiais e metodologia para ressecção apical em um modelo de mouse. As etapas mais salientes deste surviva roedorl cirurgia envolve anestesia hipotermia, toracotomia, amputação cirúrgica do ápice ventricular coração, e sutura e recuperação de ratos. A abordagem descrita poderia expandir a aplicação do modelo de rato ressecção apical para a investigação cardiovascular.
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
Os autores agradecem Drs. James Hawkins, Zu-Xi Yu Xuan Qu e do National Heart, Lung, and Blood Institute (NHLBI) por sua assistência com a cirurgia do mouse e preparação e coloração de secções de parafina. Os autores são gratos ao Conselho Editorial NIH Fellows de assistência editorial.
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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