Supported in part by NHLBI Grants P01-HL039707 and P01-HL087226 and the Leducq Foundation (J.J. and O.B.), by a Spanish Society of Cardiology Fellowship, Fundaci&oacute;n Pedro Barri&eacute; de la Maza and Fundaci&oacute;n Alfonso Mart&iacute;n Escudero (D.F.R.), by F&eacute;d&eacute;ration Fran&ccedil;aise de Cardiologie (R.P.M.), by a Heart Rhythm Society fellowship Award, The Fellowship of Japan Heart Foundation/The Japanese Society of Electrocardiology (M.Y.).

1. Heart removal and Langendorff perfusion
 Sheep weighing 35-40 Kg are anesthetized using 4-6 mg/Kg propofol and 60-100 mg/Kg of sodium pentobarbital. Hearts are removed via thoracotomy and connected to a Langendorff-perfusion system with circulating oxygenated (95% O2, 5% CO2) Tyrode's solution at a constant flow rate of 240-270 ml/min, pH 7.4 and 35.5-37.5 &deg;C. The Tyrode's composition (in mM) is: NaCl 130, KCl 4.0, MgCl2 1, CaCl2 1.8, NaHCO3 24, NaH2PO4 1.2, Glucose 5.6, and Albumin 0.04 ...

<ol>
	<li>Kannel, W. B., Wolf, P. A., Benjamin, E. J., Levy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence,+incidence,+prognosis,+and+predisposing+conditions+for+atrial+fibrillation:+population-based+estimates.">Prevalence, incidence, prognosis, and predisposing conditions for atrial fibrillation: population-based estimates.</a> Am J Cardiol. 82, (1998).</li><li>Wolf, P. A., Abbott, R. D., Kannel, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atrial+fibrillation+as+an+independent+risk+factor+for+stroke:+the+Framingham+Study.">Atrial fibrillation as an independent risk factor for stroke: the Framingham Study.</a> Stroke. 22, 983-988 (1991).</li><li>Miyasaka, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secular+trends+in+incidence+of+atrial+fibrillation+in+Olmsted+County,+Minnesota,+1980+to+2000,+and+implications+on+the+projections+for+future+prevalence.">Secular trends in incidence of atrial fibrillation in Olmsted County, Minnesota, 1980 to 2000, and implications on the projections for future prevalence.</a> Circulation. 114, 119-125 (1980).</li><li>Ravelli, F., Allessie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+atrial+dilatation+on+refractory+period+and+vulnerability+to+atrial+fibrillation+in+the+isolated+Langendorff-perfused+rabbit+heart.">Effects of atrial dilatation on refractory period and vulnerability to atrial fibrillation in the isolated Langendorff-perfused rabbit heart.</a> Circulation. 96, 1686-1695 (1997).</li><li>Gray, R. A., Pertsov, A. M., Jalife, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+and+temporal+organization+during+cardiac+fibrillation.">Spatial and temporal organization during cardiac fibrillation.</a> Nature. 392, 75-78 (1998).</li><li>Gray, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+cardiac+fibrillation.">Mechanisms of cardiac fibrillation.</a> Science. 270, 1222-1225 (1995).</li><li>Samie, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rectification+of+the+background+potassium+current:+a+determinant+of+rotor+dynamics+in+ventricular+fibrillation.">Rectification of the background potassium current: a determinant of rotor dynamics in ventricular fibrillation.</a> Circ Res. 89, 1216-1223 (2001).</li><li>Kalifa, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-atrial+pressure+increases+rate+and+organization+of+waves+emanating+from+the+superior+pulmonary+veins+during+atrial+fibrillation.">Intra-atrial pressure increases rate and organization of waves emanating from the superior pulmonary veins during atrial fibrillation.</a> Circulation. 108, 668-671 (2003).</li><li>Mandapati, R., Skanes, A., Chen, J., Berenfeld, O., Jalife, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+microreentrant+sources+as+a+mechanism+of+atrial+fibrillation+in+the+isolated+sheep+heart.">Stable microreentrant sources as a mechanism of atrial fibrillation in the isolated sheep heart.</a> Circulation. 101, 194-199 (2000).</li><li>Yamazaki, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+stretch-induced+atrial+fibrillation+in+the+presence+and+the+absence+of+adrenocholinergic+stimulation:+interplay+between+rotors+and+focal+discharges.">Mechanisms of stretch-induced atrial fibrillation in the presence and the absence of adrenocholinergic stimulation: interplay between rotors and focal discharges.</a> Heart Rhythm. 6, 1009-1017 (2009).</li><li>Rama, D. F., Jalife, J., Antzelevitch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+Underlying+Atrial+Fibrillation.">Mechanisms Underlying Atrial Fibrillation.</a> Basic Science for Clinical Electrophysiologist. 3, 141-156 (2011).</li><li>Berenfeld, O., Zaitsev, A. V., Mironov, S. F., Pertsov, A. M., Jalife, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frequency-dependent+breakdown+of+wave+propagation+into+fibrillatory+conduction+across+the+pectinate+muscle+network+in+the+isolated+sheep+right+atrium.">Frequency-dependent breakdown of wave propagation into fibrillatory conduction across the pectinate muscle network in the isolated sheep right atrium.</a> Circ Res. 90, 1173-1180 (2002).</li><li>Haissaguerre, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+initiation+of+atrial+fibrillation+by+ectopic+beats+originating+in+the+pulmonary+veins.">Spontaneous initiation of atrial fibrillation by ectopic beats originating in the pulmonary veins.</a> N Engl J Med. 339, 659-666 (1998).</li><li>Warren, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blockade+of+the+inward+rectifying+potassium+current+terminates+ventricular+fibrillation+in+the+guinea+pig+heart.">Blockade of the inward rectifying potassium current terminates ventricular fibrillation in the guinea pig heart.</a> J Cardiovasc Electrophysiol. 14, 621-631 (2003).</li><li>Claerbout, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fundamentals of Geophysical Data Processing. , (1976).</li><li>Atienza, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+dominant+frequency+mapping+and+ablation+of+dominant+frequency+sites+in+atrial+fibrillation+with+left-to-right+frequency+gradients+predicts+long-term+maintenance+of+sinus+rhythm.">Real-time dominant frequency mapping and ablation of dominant frequency sites in atrial fibrillation with left-to-right frequency gradients predicts long-term maintenance of sinus rhythm.</a> Heart Rhythm. 6, 33-40 (2009).</li></ol>

The characteristics of acute stretch-induced AF in the isolated sheep heart resemble some of the properties of human paroxysmal AF. An acute increase in intra-atrial pressure in the sheep heart allows the maintenance of AF for long periods of time, similar to a higher risk of AF observed in patients with atrial dilatation.1 The presence of left-to-right DF gradients in the sheep atria is also similar to those registered in human electrophysiological studies.16 Therefore, understanding the mechanism ...

<table border="1">
<tbody>
<tr>
<td>
Material Name
</td>
<td>
Company		
</td>
<td>
Catalogue Number
</td>
</tr>
<tr>
<td>
Euthanasia
</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>
Heparin
</td>
<td>
Sigma
</td>
<td>
H3393
</td>
</tr>
<tr>
<td>
Propofol
</td>
<td>
Abbott
</td>
<td>
5206-04-03
</td>
</tr>
<tr>
<td>
Pentobarbital
</td>
<td>
Lundbeck Inc
</td>
<td>
NDC 67386-501-55
</td>
</tr>
<tr>
<td>
Introducer 18 Gauge
</td>
<td>
Terumo
</td>
<td>
SS*FF1832
</td>
</tr>
<tr>
<td>
Cuffed endotracheal tube (9 mm)
</td>
<td>
DRE Veterinary
</td>
<td>
#9440
</td>
</tr>
<tr>
<td>
Fiber Optic Laryngoscope Case
</td>
<td>
DRE Veterinary
</td>
<td>
#991
</td>
</tr>
<tr>
<td>
Fiber Optic Blade
</td>
<td>
DRE Veterinary
</td>
<td>
#984
</td>
</tr>
<tr>
<td>
Operating Scissors
</td>
<td>
DRE Veterinary
</td>
<td>
#9702
#1944
</td>
</tr>
<tr>
<td>
Scalpel Handle #3 Solid 4&quot;
</td>
<td>
Roboz Surgical Instrument Co., Inc.
</td>
<td>
RS-9843
</td>
</tr>
<tr>
<td>
Sterile Scalpel Blades
</td>
<td>
Roboz Surgical Instrument Co., Inc.
</td>
<td>
RS-9801-10
</td>
</tr>
<tr>
<td>
Ventilation bag
</td>
<td>
Westmed
</td>
<td>
562013
</td>
</tr>
<tr>
<td>
Sims Scissors Curved Sharp/Blunt
</td>
<td>
Roboz Surgical Instrument Co., Inc.
</td>
<td>
RS-7035
</td>
</tr>
<tr>
<td>
Tissue Forceps (&times;2)
</td>
<td>
DRE Veterinary
</td>
<td>
#1895
</td>
</tr>
<tr>
<td>
KANTROWITZ Thoracic Forceps, 11&quot;&nbsp;
</td>
<td>
Biomedical Research Instruments, Inc.
</td>
<td>
34-1980
</td>
</tr>
<tr>
<td>
Finochietto Large Chest Spreader
</td>
<td>
Kapp Surgical Instrument Inc.
</td>
<td>
KS-7301
</td>
</tr>
<tr>
<td>
Thoracotomy shears
</td>
<td>
Rostfrei Solingen
</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>
Plastic tray
</td>
<td>
Nalgene
</td>
<td>
Fischer
</td>
</tr>
<tr>
<td>
Optical mapping
</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>
Bonn Scissors (&times;2)
</td>
<td>
Roboz Surgical Instrument Co., Inc
</td>
<td>
RS-5840SC
</td>
</tr>
<tr>
<td>
Surgical silk
</td>
<td>
Fischer
</td>
<td>
50-900-04214
</td>
</tr>
<tr>
<td>
Micro Dissecting Forceps
</td>
<td>
Roboz Surgical Instrument Co., Inc
</td>
<td>
RS-5130
</td>
</tr>
<tr>
<td>
Tetrapolar electrode catheters (Torq) (&times;4)
</td>
<td>
Medtronic Inc.
</td>
<td>
05580SP
</td>
</tr>
<tr>
<td>
Digital sensor. Biopac Systems transducer
</td>
<td>
Biopac Systems, Inc.
</td>
<td>
RX104A
</td>
</tr>
<tr>
<td>
Biopac Systems amplifier
</td>
<td>
Biopac Systems, Inc.
</td>
<td>
DA-100C
</td>
</tr>
<tr>
<td>
Di-4-ANEPPS
</td>
<td>
Sigma-Aldrich, St.
</td>
<td>
D8604-5mg
</td>
</tr>
<tr>
<td>
Blebbistatin
</td>
<td>
Enzo Life Science International, INC.
</td>
<td>
BML-E1315-0025
</td>
</tr>
<tr>
<td>
LittleJoe CCD video camera(&times;2)
</td>
<td>
SciMeasure Analytical Systems, Inc.
</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>
Dual-channel rigid borescope
</td>
<td>
Everest VIT, Inc.
</td>
<td>
R10-25-0-90
</td>
</tr>
<tr>
<td>
Perfusion pumps (&times;2)
</td>
<td>
Cole Parmer
</td>
<td>
GK-77920-30
</td>
</tr>
<tr>
<td>
Temperature probe
</td>
<td>
Cole Parmer
</td>
<td>
R-08491-02
</td>
</tr>
<tr>
<td>
pH meter
</td>
<td>
Fischer
</td>
<td>
01-913-806
</td>
</tr>
<tr>
<td>
Digital temperature gauge
</td>
<td>
Cole Parmer
</td>
<td>
GK89000-10
</td>
</tr>
<tr>
<td>
Oxygenator filters
</td>
<td>
Sorin
</td>
<td>
05318
</td>
</tr>
<tr>
<td>
Silicon perfusion tubes (L/S 15)
</td>
<td>
MasterFlex
</td>
<td>
96410-15
</td>
</tr>
<tr>
<td>
Laser light guides (&times;6)
</td>
<td>
Oriel Corporation
</td>
<td>
77536
</td>
</tr>
<tr>
<td>
Liquid light-guide (0.2 in core)
</td>
<td>
Newport Corporation
</td>
<td>
77556
</td>
</tr>
<tr>
<td>
Laser generator (1 watt) 

(&times;1)
</td>
<td>
Shanghai Dream Lsaer Tecchnology
</td>
<td>
SDL-532-1000T
</td>
</tr>
<tr>
<td>
Laser generator (5 watt)
 (&times;1)
</td>
<td>
Spectra Physics Lasers
</td>
<td>
MILL 5sJ
</td>
</tr>
</tbody>
</table>

high-resolution endocardial and epicardial optical mapping in a sheep model of stretch-induced atrial fibrillation

Atrial fibrillation (AF) is a complex cardiac arrhythmia with high morbidity and mortality.1,2 It is the most common sustained cardiac rhythm disturbance seen in clinical practice and its prevalence is expected to increase in the coming years.3 Increased intra-atrial pressure and dilatation have been long recognized to lead to AF,1,4 which highlights the relevance of using animal models and stretch to study AF dynamics. Understanding the mechanisms underlying AF requires visualization of the cardiac electrical waves with high spatial and temporal resolution. While high-temporal resolution can be achieved by conventional electrical mapping traditionally used in human electrophysiological studies, the small number of intra-atrial electrodes that can be used simultaneously limits the spatial resolution and precludes any detailed tracking of the electrical waves during the arrhythmia. The introduction of optical mapping in the early 90's enabled wide-field characterization of fibrillatory activity together with sub-millimeter spatial resolution in animal models5,6 and led to the identification of rapidly spinning electrical wave patterns (rotors) as the sources of the fibrillatory activity that may occur in the ventricles or the atria.7-9 Using combined time- and frequency-domain analyses of optical mapping it is possible to demonstrate discrete sites of high frequency periodic activity during AF, along with frequency gradients between left and right atrium. The region with fastest rotors activates at the highest frequency and drives the overall arrhythmia.10,11 The waves emanating from such rotor interact with either functional or anatomic obstacles in their path, resulting in the phenomenon of fibrillatory conduction.12 Mapping the endocardial surface of the posterior left atrium (PLA) allows the tracking of AF wave dynamics in the region with the highest rotor frequency. Importantly, the PLA is the region where intracavitary catheter-based ablative procedures are most successful terminating AF in patients,13 which underscores the relevance of studying AF dynamics from the interior of the left atrium. Here we describe a sheep model of acute stretch-induced AF, which resembles some of the characteristics of human paroxysmal AF. Epicardial mapping on the left atrium is complemented with endocardial mapping of the PLA using a dual-channel rigid borescope c-mounted to a CCD camera, which represents the most direct approach to visualize the patterns of activation in the most relevant region for AF maintenance.

Watch this Scientific Journal Video about High-Resolution Endocardial and Epicardial Optical Mapping in a Sheep Model of Stretch-Induced Atrial Fibrillation at JoVE.com

High-Resolution Endocardial and Epicardial Optical Mapping in a Sheep Model of Stretch-Induced Atrial Fibrillation

This report provides a detailed description of the methodology and results of simultaneous endocardial and epicardial optical mapping of electrical excitation in the intact left atrium of a Langendorff-perfused sheep heart during stretch-induced atrial fibrillation.

 Atrial fibrillation (AF) is a complex cardiac arrhythmia with high morbidity and mortality.1,2 It is the most common sustained cardiac rhythm disturbance ...

high-resolution-endocardial-epicardial-optical-mapping-sheep-model

Research

JoVE Journal

Medicine

14.7K ビュー.  University of Michigan. このレポートは、ストレッチ誘発性心房細動時のランゲンドルフ灌流羊の心臓の無傷の左心房の電気的興奮を同時に心内膜と心外膜の光学的マッピングの方法論と結果の詳細な説明を提供します。

ビデオ: High-Resolution Endocardial and Epicardial Optical Mapping in a Sheep Model of Stretch-Induced Atrial Fibrillation

This report provides a detailed description of the methodology and results of simultaneous endocardial and epicardial optical mapping of electrical excitation in the intact left atrium of a Langendorff-perfused sheep heart during stretch-induced atrial...

high-resolution optical mapping of the mouse sino-atrial node

High-resolution Optical Mapping of the Mouse Sino-atrial Node

Here, we present a protocol for optical mapping of electrical activity from the mouse right atrium and especially the sino-atrial node, at a high spatial and temporal...

electrophysiological assessment of murine atria with high-resolution optical mapping

Electrophysiological Assessment of Murine Atria with High-Resolution Optical Mapping

This protocol describes the electrophysiological evaluation of murine atria utilizing an optical mapping system with a high temporal and spatial resolution, including dual recordings of the membrane voltage and Ca2+ transient under programmed stimulation through a specialized electrode...

sterile pericarditis in aachener minipigs as a model for atrial myopathy and atrial fibrillation

Sterile Pericarditis in Aachener Minipigs As a Model for Atrial Myopathy and Atrial Fibrillation

We describe a sterile pericarditis model in minipigs to study atrial myopathy and atrial fibrillation (AF). We present surgical and anesthetic techniques, strategies for vascular access, and a protocol to study the inducibility of...

optimization of transesophageal atrial pacing to assess atrial fibrillation susceptibility in mice

Optimization of Transesophageal Atrial Pacing to Assess Atrial Fibrillation Susceptibility in Mice

The present protocol describes the optimization of experimental parameters when using transesophageal atrial pacing to assess atrial fibrillation susceptibility in...

transesophageal atrial burst pacing for atrial fibrillation induction in rats

Transesophageal Atrial Burst Pacing for Atrial Fibrillation Induction in Rats

The present work describes an experimental protocol of transesophageal atrial burst pacing for efficient induction of atrial fibrillation (AF) in rats. The protocol can be used in rats with healthy or remodeled hearts, allowing the study of AF pathophysiology, identification of novel therapeutic targets, and evaluation of new therapeutic...

voltage and calcium dual channel optical mapping of cultured hl-1 atrial myocyte monolayer

Voltage and Calcium Dual Channel Optical Mapping of Cultured HL-1 Atrial Myocyte Monolayer

This article describes the technique used to perform dual channel optical mapping in cultured HL-1 atrial cell monolayers. This unique protocol allows the simultaneous visualization of both calcium (Ca) and voltage (Vm) activity in the same area for the detailed detection and analysis of electrophysiological properties of culture...

robotic ablation of atrial fibrillation

Robotic Ablation of Atrial Fibrillation

Pulmonary vein isolation (PVI) with an ablation catheter is a curative treatment for atrial fibrillation (AF). Robotic catheter systems aim to improve catheter steerability. Here, a procedure with a new robotic catheter system is presented. The goal of the procedure is electrical block between pulmonary vein and left...

estimating bilateral atrial function by cardiovascular magnetic resonance feature tracking in patients with paroxysmal atrial fibrillation

Estimating Bilateral Atrial Function by Cardiovascular Magnetic Resonance Feature Tracking in Patients with Paroxysmal Atrial Fibrillation

The atrial function is associated with the strain and strain rate. The cardiac magnetic resonance feature tracking (CMR-FT) technique was used in this study to quantify left and right atrial global and segmental longitudinal strain and strain rate in individuals with paroxysmal atrial...

catheter ablation in combination with left atrial appendage closure for atrial fibrillation

Catheter Ablation in Combination With Left Atrial Appendage Closure for Atrial Fibrillation

Catheter ablation is combined with placement of the WATCHMAN Left Atrial Appendage Closure Device to prevent ischemic stroke in a patient with non-valvular atrial...

high-throughput analysis of optical mapping data using electromap

High-Throughput Analysis of Optical Mapping Data Using ElectroMap

This protocol describes the setup and use of ElectroMap, a MATLAB-based open-source software platform for analysis of cardiac optical mapping data. ElectroMap provides a versatile high-throughput tool for analysis of optical mapping voltage and calcium datasets across a wide range of cardiac experimental...

a model of long-term ventricular fibrillation in isolated rat hearts

A Model of Long-Term Ventricular Fibrillation in Isolated Rat Hearts

This protocol presents a model of long-term ventricular fibrillation in rat hearts induced by continuous stimulation with low-voltage alternating current. This model has a high success rate, is stable, reliable, and reproducible, has a low impact on cardiac function, and causes only mild myocardial...

method for high speed stretch injury of human induced pluripotent stem cell-derived neurons in a 96-well format

Method for High Speed Stretch Injury of Human Induced Pluripotent Stem Cell-derived Neurons in a 96-well Format

Here we present a method for a human in vitro model of stretch injury in a 96-well format on a timescale relevant to impact trauma. This includes methods for fabricating stretchable plates, quantifying the mechanical insult, culturing and injuring cells, imaging, and high content analysis to quantify...

construction of a high resolution microscope with conventional and holographic optical trapping capabilities

Construction of a High Resolution Microscope with Conventional and Holographic Optical Trapping Capabilities

The system described herein employs a traditional optical trap as well as an independent holographic optical trapping line, capable of creating and manipulating multiple traps. This allows for the creation of complex geometric arrangements of refractive particles while also permitting simultaneous high-speed, high-resolution measurements of the activity of biological...

the watchman left atrial appendage closure device for atrial fibrillation

The WATCHMAN Left Atrial Appendage Closure Device for Atrial Fibrillation

The accompanying video describes a procedure for percutaneous placement of the WATCHMAN Left Atrial Appendage (LAA) Device.  The WATCHMAN is a nitinol device designed to be permanently implanted at, or slightly distal to, the opening of the left atrial appendage (LAA) to trap blood clots before they exit the LAA, preventing thromboembolic...

a rat model of ventricular fibrillation and resuscitation by conventional closed-chest technique

A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation...

optical mapping of langendorff-perfused rat hearts

Optical Mapping of Langendorff-perfused Rat Hearts

This article describes a high temporal and spatial resolution technique to optically image action potential movement on the surface of Langendorff-perfused rat hearts using a potentiometric dye...

fiber optic distributed sensors for high-resolution temperature field mapping

Fiber Optic Distributed Sensors for High-resolution Temperature Field Mapping

We demonstrate use of a fiber optic distributed sensor for mapping the temperature field of mixing air jets. The Rayleigh scattering-based sensor generates thousands of data points along a single fiber to provide exceptional spatial resolution that is unattainable with traditional sensors such as...

optical mapping of action potentials and calcium transients in the mouse heart

Optical Mapping of Action Potentials and Calcium Transients in the Mouse Heart

This paper details the dissection procedure, instrumental setup, and experimental conditions during optical mapping of transmembrane potential (Vm) and intracellular calcium transient (CaT) in intact isolated Langendorff perfused mouse...

cooling or warming the esophagus to reduce esophageal injury during left atrial ablation in the treatment of atrial fibrillation

Cooling or Warming the Esophagus to Reduce Esophageal Injury During Left Atrial Ablation in the Treatment of Atrial Fibrillation

The goal of this protocol is to describe the use of esophageal temperature modulation to counteract esophageal thermal injury from left atrial ablation for the treatment of atrial...

Atrial fibrillation (AF) is the most common cardiac arrhythmia, affecting an estimated 6 million people in the United States 1. Since AF affects primarily elderly people, its prevalence increases parallel with age. As such, it is expected that 15.9 million Americans will be affected by the year 2050 2. Ischemic stroke occurs in 5% of non-anticoagulated AF patients each year. Current treatments for AF include rate control, rhythm control and prevention of stroke 3.
The American College of Cardiology, American Heart Association, and European Society of Cardiology currently recommended rate control as the first course of therapy for AF 3. Rate control is achieved by administration of pharmacological agents, such as &beta;-blockers, that lower the heart rate until it reaches a less symptomatic state 3. Rhythm control aims to return the heart to its normal sinus rhythm and is typically achieved through administration of antiarrhythmic drugs such as amiodarone, electrical cardioversion or ablation therapy. Rhythm control methods, however, have not been demonstrated to be superior to rate-control methods 4-6. In fact, certain antiarrhythmic drugs have been shown to be associated with higher hospitalization rates, serious adverse effects 3, or even increases in mortality in patients with structural heart defects 7. Thus, treatment with antiarrhythmics is more often used when rate-control drugs are ineffective or contraindicated. Rate-control and antiarrhythmic agents relieve the symptoms of AF, including palpitations, shortness of breath, and fatigue 8, but don't reliably prevent thromboembolic events 6.
Treatment with the anticoagulant drug warfarin significantly reduces the rate of stroke or embolism 9,10. However, because of problems associated with its use, fewer than 50% of patients are treated with it. The therapeutic dose is affected by drug, dietary, and metabolic interactions, and thus requires detailed monitoring. In addition, warfarin has the potential to cause severe, sometimes lethal, bleeding 2. As an alternative, aspirin is commonly prescribed. While aspirin is typically well tolerated, it is far less effective at preventing stroke 10. Other alternatives to warfarin, such as dabigatran 11 or rivaroxaban 12 demonstrate non-inferiority to warfarin with respect to thromboembolic events (in fact, dabigatran given as a high dose of 150 mg twice a day has shown superiority). While these drugs have the advantage of eliminating dietary concerns and eliminating the need for regular blood monitoring, major bleeding and associated complications, while somewhat less so than with warfarin, remain an issue 13-15.
Since 90% of AF-associated strokes result from emboli that arise from the left atrial appendage (LAA) 2, one alternative approach to warfarin therapy has been to exclude the LAA using an implanted device to trap blood clots before they exit. Here, we demonstrate a procedure for implanting the WATCHMAN Left Atrial Appendage Closure Device. A transseptal cannula is inserted through the femoral vein, and under fluoroscopic guidance, inter-atrial septum is crossed. Once access to the left atrium has been achieved, a guidewire is placed in the upper pulmonary vein and the WATCHMAN Access Sheath and dilator are advanced over the wire into the left atrium. The guidewire is removed, and the access sheath is carefully advanced into the distal portion of the LAA over a pigtail catheter. The WATCHMAN Delivery System is prepped, inserted into the access sheath, and slowly advanced. The WATCHMAN device is then deployed into the LAA. The device release criteria are confirmed via fluoroscopy and transesophageal echocardiography (TEE) and the device is released.

The accompanying video describes a procedure for percutaneous placement of the WATCHMAN Left Atrial Appendage (LAA) Device. The WATCHMAN is a nitinol device designed to be permanently implanted at, or slightly distal to, the opening of the left atrial appendage (LAA) to trap blood clots before they exit the LAA, preventing thromboembolic stroke.

Atrial fibrillation (AF) is the most common cardiac arrhythmia, affecting an estimated 6 million people in the United States 1. Since AF affects primarily ...

Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia, affecting millions of individuals worldwide 1-3. The rapid, irregular, and disordered electrical activity in the atria gives rise to palpitations, fatigue, dyspnea, chest pain and dizziness with or without syncope 4, 5. Patients with AF have a five-fold higher risk of stroke 6.
Oral anticoagulation (OAC) with warfarin is commonly used for stroke prevention in patients with AF and has been shown to reduce the risk of stroke by 64% 7. Warfarin therapy has several major disadvantages, however, including bleeding, non-tolerance, interactions with other medications and foods, non-compliance and a narrow therapeutic range 8-11. These issues, together with poor appreciation of the risk-benefit ratio, unawareness of guidelines, or absence of an OAC monitoring outpatient clinic may explain why only 30-60% of patients with AF are prescribed this drug 8.
The problems associated with warfarin, combined with the limited efficacy and/or serious side effects associated with other medications used for AF 12,13, highlight the need for effective non-pharmacological approaches to treatment. One such approach is catheter ablation (CA), a procedure in which a radiofrequency electrical current is applied to regions of the heart to create small ablation lesions that electrically isolate potential AF triggers 4. CA is a well-established treatment for AF symptoms 14, 15, that may also decrease the risk of stroke. Recent data showed a significant decrease in the relative risk of stroke and transient ischemic attack events among patients who underwent ablation compared with those undergoing antiarrhythmic drug therapy 16.
Since the left atrial appendage (LAA) is the source of thrombi in more than 90% of patients with non-valvular atrial fibrillation 17, another approach to stroke prevention is to physically block clots from exiting the LAA. One method for occluding the LAA is via percutaneous placement of the WATCHMAN LAA closure device. The WATCHMAN device resembles a small parachute. It consists of a nitinol frame covered by fabric polyethyl terephthalate that prevents emboli, but not blood, from exiting during the healing process. Fixation anchors around the perimeter secure the device in the LAA (Figure 1). To date, the WATCHMAN is the only implanted percutaneous device for which a randomized clinical trial has been reported. In this study, implantation of the WATCHMAN was found to be at least as effective as warfarin in preventing stroke (all-causes) and death (all-causes) 18. This device received the Conformit&eacute; Europ&eacute;enne (CE) mark for use in the European Union for warfarin eligible patients and in those who have a contraindication to anticoagulation therapy 19.
Given the proven effectiveness of CA to alleviate AF symptoms and the promising data with regard to reduction of thromboembolic events with both CA and WATCHMAN implantation, combining the two procedures is hoped to further reduce the incidence of stroke in high-risk patients while simultaneously relieving symptoms. The combined procedure may eventually enable patients to undergo implantation of the WATCHMAN device without subsequent warfarin treatment, since the CA procedure itself reduces thromboembolic events. This would present an avenue of treatment previously unavailable to patients ineligible for warfarin treatment because of recurrent bleeding 20 or other warfarin-associated problems.
The combined procedure is performed under general anesthesia with biplane fluoroscopy and TEE guidance. Catheter ablation is followed by implantation of the WATCHMAN LAA closure device. Data from a non-randomized trial with 10 patients demonstrates that this procedure can be safely performed in patients with a CHADS2 score of greater than 1 21. Further studies to examine the effectiveness of the combined procedure in reducing symptoms from AF and associated stroke are therefore warranted.

Catheter ablation is combined with placement of the WATCHMAN Left Atrial Appendage Closure Device to prevent ischemic stroke in a patient with non-valvular atrial fibrillation.

Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia, affecting millions of individuals worldwide 1-3. The rapid, irregular, and ...

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic components of cardiopulmonary resuscitation in humans is herein described. The model was developed in 1988 and has been used in approximately 70 peer-reviewed publications examining a myriad of resuscitation aspects including its physiology and pathophysiology, determinants of resuscitability, pharmacologic interventions, and even the effects of cell therapies. The model featured in this presentation includes: (1) vascular catheterization to measure aortic and right atrial pressures, to measure cardiac output by thermodilution, and to electrically induce ventricular fibrillation; and (2) tracheal intubation for positive pressure ventilation with oxygen enriched gas and assessment of the end-tidal CO2. A typical sequence of intervention entails: (1) electrical induction of ventricular fibrillation, (2) chest compression using a mechanical piston device concomitantly with positive pressure ventilation delivering oxygen-enriched gas, (3) electrical shocks to terminate ventricular fibrillation and reestablish cardiac activity, (4) assessment of post-resuscitation hemodynamic and metabolic function, and (5) assessment of survival and recovery of organ function. A robust inventory of measurements is available that includes &#8211; but is not limited to &#8211; hemodynamic, metabolic, and tissue measurements. The model has been highly effective in developing new resuscitation concepts and examining novel therapeutic interventions before their testing in larger and translationally more relevant animal models of cardiac arrest and resuscitation.

This article describes a rat model of electrically-induced ventricular fibrillation and resuscitation by chest compression, ventilation, and delivery of electrical shocks that simulates an episode of sudden cardiac arrest and conventional cardiopulmonary resuscitation. The model enables gathering insights on the pathophysiology of cardiac arrest and exploration of new resuscitation strategies.

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic ...

Background: Pulmonary vein isolation (PVI) is an established treatment for atrial fibrillation (AF). During PVI an electrical conduction block between pulmonary vein (PV) and left atrium (LA) is created. This conduction block prevents AF, which is triggered by irregular electric activity originating from the PV. However, transmural atrial lesions are required which can be challenging. Re-conduction and AF recurrence occur in 20 - 40% of the cases. Robotic catheter systems aim to improve catheter steerability. Here, a procedure with a new remote catheter system (RCS), is presented. Objective of this article is to show feasibility of robotic AF ablation with a novel system. Materials and Methods: After interatrial trans-septal puncture is performed using a long sheath and needle under fluoroscopic guidance. The needle is removed and a guide wire is placed in the left superior PV. Then an ablation catheter is positioned in the LA, using the sheath and wire as guide to the LA. LA angiography is performed over the sheath. A circular mapping catheter is positioned via the long sheath into the LA and a three-dimensional (3-D) anatomical reconstruction of the LA is performed. The handle of the ablation catheter is positioned in the robotic arm of the Amigo system and the ablation procedure begins. During the ablation procedure, the operator manipulates the ablation catheter via the robotic arm with the use of a remote control. The ablation is performed by creating point-by-point lesions around the left and right PV ostia. Contact force is measured at the catheter tip to provide feedback of catheter-tissue contact. Conduction block is confirmed by recording the PV potentials on the circular mapping catheter and by pacing maneuvers. The operator stays out of the radiationfield during ablation. Conclusion: The novel catheter system allows ablation with high stability on low operator fluoroscopy exposure.

Pulmonary vein isolation (PVI) with an ablation catheter is a curative treatment for atrial fibrillation (AF). Robotic catheter systems aim to improve catheter steerability. Here, a procedure with a new robotic catheter system is presented. The goal of the procedure is electrical block between pulmonary vein and left atrium.

Background: Pulmonary vein isolation (PVI) is an established treatment for atrial fibrillation (AF). During PVI an electrical conduction block between ...

Recent genome-wide association studies targeting atrial fibrillation (AF) have indicated a strong association between the genotype and electrophysiological phenotype in the atria. That encourages us to utilize a genetically-engineered mouse model to elucidate the mechanism of AF. However, it is difficult to evaluate the electrophysiological properties in murine atria due to their small size. This protocol describes the electrophysiological evaluation of atria using an optical mapping system with a high temporal and spatial resolution in Langendorff perfused murine hearts. The optical mapping system is assembled with dual high-speed complementary metal oxide semiconductor cameras and high magnification objective lenses, to detect the fluorescence of a voltage-sensitive dye and Ca2+ indicator. To focus on the assessment of murine atria, optical mapping is performed with an area of 2 mm &#215; 2 mm or 10 mm x 10 mm, with a 100 &#215; 100 resolution (20 &#181;m/pixel or 100 &#181;m/pixel) and sampling rate of up to 10 kHz (0.1 ms) at maximum. A 1-French size quadripolar electrode pacing catheter is placed into the right atrium through the superior vena cava avoiding any mechanical damage to the atrium, and pacing stimulation is delivered through the catheter. An electrophysiological study is performed with programmed stimulation including constant pacing, burst pacing, and up to triple extrastimuli pacing. Under a spontaneous or pacing rhythm, the optical mapping recorded the action potential duration, activation map, conduction velocity, and Ca2+ transient individually in the right and left atria. In addition, the programmed stimulation also determines the inducibility of atrial tachyarrhythmias. Precise activation mapping is performed to identify the propagation of the excitation in the atrium during an induced atrial tachyarrhythmia. Optical mapping with a specialized setting enables a thorough electrophysiological evaluation of the atrium in murine pathological models.

This protocol describes the electrophysiological evaluation of murine atria utilizing an optical mapping system with a high temporal and spatial resolution, including dual recordings of the membrane voltage and Ca2+ transient under programmed stimulation through a specialized electrode catheter.

Recent genome-wide association studies targeting atrial fibrillation (AF) have indicated a strong association between the genotype and ...

Optical mapping is an established technique for high spatio-temporal resolution study of cardiac electrophysiology in multi-cellular preparations. Here we present, in a step-by-step guide, the use of ElectroMap for analysis, quantification, and mapping of high-resolution voltage and calcium datasets acquired by optical mapping. ElectroMap analysis options cover a wide variety of key electrophysiological parameters, and the graphical user interface allows straightforward modification of pre-processing and parameter definitions, making ElectroMap applicable to a wide range of experimental models. We show how built-in pacing frequency detection and signal segmentation allows high-throughput analysis of entire experimental recordings, acute responses, and single beat-to-beat variability. Additionally, ElectroMap incorporates automated multi-beat averaging to improve signal quality of noisy datasets, and here we demonstrate how this feature can help elucidate electrophysiological changes that might otherwise go undetected when using single beat analysis. Custom modules are included within the software for detailed investigation of conduction, single file analysis, and alternans, as demonstrated here. This software platform can be used to enable and accelerate the processing, analysis, and mapping of complex cardiac electrophysiology.

This protocol describes the setup and use of ElectroMap, a MATLAB-based open-source software platform for analysis of cardiac optical mapping data. ElectroMap provides a versatile high-throughput tool for analysis of optical mapping voltage and calcium datasets across a wide range of cardiac experimental models.

Optical mapping is an established technique for high spatio-temporal resolution study of cardiac electrophysiology in multi-cellular preparations. Here we ...

Ablation of the left atrium using either radiofrequency (RF) or cryothermal energy is an effective treatment for atrial fibrillation (AF) and is the most frequent type of cardiac ablation procedure performed. Although generally safe, collateral injury to surrounding structures, particularly the esophagus, remains a concern. Cooling or warming the esophagus to counteract the heat from RF ablation, or the cold from cryoablation, is a method that is used to reduce thermal esophageal injury, and there are increasing data to support this approach. This protocol describes the use of a commercially available esophageal temperature management device to cool or warm the esophagus to reduce esophageal injury during left atrial ablation. The temperature management device is powered by standard water-blanket heat exchangers, and is shaped like a standard orogastric tube placed for gastric suctioning and decompression. Water circulates through the device in a closed-loop circuit, transferring heat across the silicone walls of the device, through the esophageal wall. Placement of the device is analogous to the placement of a typical orogastric tube, and temperature is adjusted via the external heat-exchanger console.

The goal of this protocol is to describe the use of esophageal temperature modulation to counteract esophageal thermal injury from left atrial ablation for the treatment of atrial fibrillation.

Ablation of the left atrium using either radiofrequency (RF) or cryothermal energy is an effective treatment for atrial fibrillation (AF) and is the most ...

Atrial fibrillation (AF) is the most common arrhythmia caused by structural remodeling of the atria, also called atrial myopathy. Current therapies only target the electrical abnormalities and not the underlying atrial myopathy. For the development of novel therapies, a reproducible large animal model of atrial myopathy is necessary. This paper presents a model of sterile pericarditis-induced atrial myopathy in Aachener minipigs. Sterile pericarditis was induced by spraying sterile talcum and leaving a layer of sterile gauze over the atrial epicardial surface. This led to inflammation and fibrosis, two crucial components of the pathophysiology of atrial myopathy, making the atria susceptible to the induction of AF. Two pacemaker electrodes were positioned epicardially on each atrium and connected to two pacemakers from different manufacturers. This strategy allowed for repeated non-invasive atrial programmed stimulation to determine the inducibility of AF at specified time points after surgery. Different protocols to test AF inducibility were used. The advantages of this model are its clinical relevance, with AF inducibility and the rapid induction of inflammation and fibrosis-both present in atrial myopathy-and its reproducibility. The model will be useful in the development of novel therapies targeting atrial myopathy and AF.

We describe a sterile pericarditis model in minipigs to study atrial myopathy and atrial fibrillation (AF). We present surgical and anesthetic techniques, strategies for vascular access, and a protocol to study the inducibility of AF.

Atrial fibrillation (AF) is the most common arrhythmia caused by structural remodeling of the atria, also called atrial myopathy. Current therapies only ...

High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering from its most severe form, chronic limb-threatening ischemia (CLTI), face substantial impairments to daily living, including chronic pain, limited walking distance without pain, and nonhealing wounds. Preclinical models have been developed in various animals to study PAD, but mouse hindlimb ischemia remains the most widely used. There can be significant variation in response to ischemic insult in these models depending on the mouse strain used and the site, number, and means of arterial disruption. This protocol describes a unique method combining femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase (NOS) inhibitor to reliably induce footpad gangrene in Friend Virus B (FVB) mice that resembles the tissue loss of CLTI. While traditional means of assessing reperfusion such as laser Doppler perfusion imaging (LDPI) are still recommended, intracardiac perfusion of the lipophilic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) is used to label the vasculature. Subsequent whole-mount confocal laser scanning microscopy allows for high-resolution, three-dimensional (3D) reconstruction of footpad vascular networks that complements traditional means of assessing reperfusion in hindlimb ischemia models.

The present protocol describes a unique, clinically relevant model of peripheral arterial disease that combines femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase inhibitor to induce hindlimb gangrene in FVB mice. Intracardiac DiI perfusion is then used for high-resolution, three-dimensional imaging of the footpad vasculature.

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering ...

Animal studies have brought important insights into our understanding regarding atrial fibrillation (AF) pathophysiology and therapeutic management. Reentry, one of the main mechanisms involved in AF pathogenesis, requires a certain mass of myocardial tissue in order to occur. Due to the small size of the atria, rodents have long been considered 'resistant' to AF. Although spontaneous AF has been shown to occur in rats, long-term follow-up (up to 50 weeks) is required for the arrhythmia to occur in those models. The present work describes an experimental protocol of transesophageal atrial burst pacing for rapid and efficient induction of AF in rats. The protocol can be successfully used in rats with healthy or remodeled hearts, in the presence of a wide variety of risk factors, allowing the study of AF pathophysiology, identification of novel therapeutic targets, and evaluation of novel prophylactic and/or therapeutic strategies.

The present work describes an experimental protocol of transesophageal atrial burst pacing for efficient induction of atrial fibrillation (AF) in rats. The protocol can be used in rats with healthy or remodeled hearts, allowing the study of AF pathophysiology, identification of novel therapeutic targets, and evaluation of new therapeutic strategies.

Animal studies have brought important insights into our understanding regarding atrial fibrillation (AF) pathophysiology and therapeutic management. ...