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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Disclosures
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

The present protocol describes a modified and simplified technique with a minimally invasive transverse aortic constriction (TAC) procedure using a self-made retractor. This procedure can be conducted without a ventilator or microscope and introduces pressure overload, eventually leading to cardiac hypertrophy or heart failure.

Abstract

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of pressure overload in mouse models. The main challenge of this procedure is to clearly visualize the transverse aortic arch and precisely band the target vessel. Classical approaches perform a partial thoracotomy to expose the transverse aortic arch. However, it is an open-chest model that causes a rather large surgical trauma and requires a ventilator during the surgery. To prevent unnecessary trauma and simplify the operating procedure, the aortic arch is approached via the proximal proportion of the sternum, reaching and binding the target vessel using a small self-made retractor that contains a snare. This procedure can be conducted without entering the pleura cavity and does not need a ventilator or microsurgical operation, which leaves the mice with physiological breathing patterns, simplifies the procedure, and significantly reduces operation time. Due to the less invasive approach and less operation time, mice can undergo fewer stress reactions and recover rapidly.

Introduction

Heart failure is a complex clinical symptom that results from impaired structure and function of ventricular filling or ejecting blood1. The disease stage is mainly defined via the New York Heart Association function classification based on the severity of symptoms and physical activity2. For those patients with an ejection fraction of over 50%, structural and/or functional abnormalities raised natriuretic peptides to support the diagnosis of heart failure with preserved ejection fraction (HFpEF)2. Ischemic heart disease is a leading cause among multiple etiologies of heart failure. Thus,....

Protocol

The current protocol is approved by the ethics committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. This procedure is performed on male adult C57/BL6 mice (>10 weeks of age). All surgical instruments were sterilized by autoclaving before the operation.

1. Preparation of surgical instrument

  1. Prepare a 5 mL syringe and pinch off the tip of the needle with a needle holder to blunt it. 
  2. Prepare a 27 G needle and blunt it with a needle holder. Cutting the tip of the needle with ophthalmic scissors is an alternative way to blunt the needle....

Representative Results

After successful TAC surgery, pressure overload was detected using an ultrasound imaging system. Four weeks after surgery, mice develop decreased heart function. In the present study, the efficacy of TAC surgery was validated via ejection fraction (EF), fractional shortening (FS), left ventricular mass (LV mass) and left ventricular internal diameter (LVID) of mice who underwent TAC surgery after 4 weeks. EF was significantly reduced in TAC mice after 4 weeks compared to sham mice (47% ± 10% vs. 78% ± .......

Discussion

The induction of sustained pressure overload can gradually cause cardiac hypertrophy and heart failure. This model has been used in numerous laboratories around the world14,15,16. The protocol provided an improved TAC method that does not need microsurgical skills or mechanical ventilation.

The most important step in this protocol is passing silk suture under the aortic arch. When the snare has hooked.......

Disclosures

The authors declared that there are no conflicts of interest.

Acknowledgements

This work is funded by the National Natural Science Foundation of China (NSFC 81822002). We thank all the members who took part in this work.

....

Materials

NameCompanyCatalog NumberComments
4-0 nonabsorbable sutureJinhuanHM403Used for suturing the skin
5 mL syringeHaifuda Technology Co., Ltd.BD-309628Used for making snare containing retractor
7-0 nonabsorbable sutureJinhuanHM701Used for aorta ligation
Animal temperature monitorKaerwenFT3400Used for monitoring body temperature
Buprenorphine SigmaB-044Used for post-surgical pain treatment
Depilatory cream VeetN/AUsed for remove body hair from the surgical area
Heating PadXiaochuangxinN/AUsed for maintaining body temperature
IbuprofenMCEHY-78131Used for post-surgical pain treatment
Iron wire (0.5 mm)Qing YuanIron wire #26Used for making snare containing retractor
Microscopic tweezersRWDF12006-10Used for penetrating and separating the tissue to open operation space
Needle holderRWDF12005-10Used for pinching off the tip of gauge needle and blunting it
Ophthalmic forcepsRWDF14012-10 Used for holding skin and other tissues
Ophthalmic scissorsRWDS11001-08Used for making sking incision of mouse
Pentobarbital sodiumSigmaP3761Used for mouse anesthesia
Sterile operating matHale & hearty211002Used for placing animal during surgery
Ultra-sound imaging systemFujifilm visualsonicsvevo1100Used for measure the blood flow velocity, left ventricular wall thickness and ejection fraction, https://www.visualsonics.com/product/imaging-systems/vevo-1100

References

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Transverse Aortic ConstrictionTAC SurgeryMouse ModelSurgical TechniqueAortic ArchSelf made RetractorNeedle SnareMinimally InvasiveAnesthesiaSterile Technique

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