Name: a murine closed-chest model of myocardial ischemia and reperfusion
Uploaded: 2012-07-17T12:00:00
Description: Watch this Scientific Journal Video about A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion at JoVE.com

We thank Daniel Duerr for his advice regarding perfusion-fixation technique.

1. Induction of Anesthesia
<ol>
	<li>For induction with isoflurane, place the mouse into an induction box which is connected to the vapor set to 3.0 Vol% and oxygen flow of 0.5 L/min.</li>
	<li>After unconsciousness is achieved with tactile stimulus failing to induce a response and the forelimb or hindlimb pedal withdrawal reflex being absent, place the mouse on a temperature-controlled operating table in a supine position. Maintain anesthesia over a nasal cone which is connected to the vapor via the induction box. Rec...

<ol>
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We have modified a murine closed-chest model by accessing the heart through a left lateral intercostal thoracotomy and leading out the LAD sutures to the chest in the left midclavicular line. Leaving the bony rib cage intact will minimize trauma, need for pain medication, surgical site infection and thus, facilitate recovery. By preserving the left internal mammal artery there is no need for electrocautery. We leave the suture loop in the subcutaneous tissue for later easy access and use a hanging weight system fo...

<table border="1">
<tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>Vapor</td>
 <td>Dr&auml;gerwerk AG</td>
 <td>Isoflo</td>
 </tr>
 <tr>
 <td>Microscope</td>
 <td>Leica</td>
 <td>M80</td>
 </tr>
 <tr>
 <td>Light source</td>
 <td>Schott</td>
 <td>KL 1500 LCD</td>
 </tr>
 <tr>
 <td>Homeothermic Blanket Control Unit</td>
 <td>Harvard Apparatus</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>MiniVent Type 845</td>
 <td>Hugo Sachs Elektronik</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>8-0 Prolene</td>
 <td>Ethicon</td>
 <td>BV130-5 6.5mm 3/8c</td>
 </tr>
 <tr>
 <td>6-0 Prolene</td>
 <td>Ethicon</td>
 <td>BV-1 9.3 mm 3/8c</td>
 </tr>
 <tr>
 <td>Kalt suture needle size 3</td>
 <td>FST</td>
 <td>12050-03</td>
 </tr>
 <tr>
 <td>Triphenyltetrazolium</td>
 <td>Sigma Aldrich</td>
 <td>93145</td>
 </tr>
 <tr>
 <td>Phthalo blue</td>
 <td>Heucotech LTD</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>PowerLab</td>
 <td>ADInstruments</td>
 <td>&nbsp;</td>
 </tr>
</tbody> 
</table>

a murine closed-chest model of myocardial ischemia and reperfusion

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ischemia and reperfusion. Immune response includes cytokine expression and release or secretion of endogenous ligands of innate immune receptors. Activation of innate immunity can potentially modulate infarct size. We have modified an existing murine closed-chest model using hanging weights which could be useful for studying myocardial pre- and postconditioning and the role of innate immunity in myocardial ischemia and reperfusion. This model allows animals to recover from surgical trauma before onset of myocardial ischemia.
Volatile anesthetics have been intensely studied and their preconditioning effect for the ischemic heart is well known. However, this protective effect precludes its use in open chest models of coronary artery ligation. Thus, another advantage could be the use of the well controllable volatile anesthetics for instrumentation in a chronic closed-chest model, since their preconditioning effect lasts up to 72 hours. Chronic heart diseases with intermittent ischemia and multiple hit models are other possible applications of this model.
For the chronic closed-chest model, intubated and ventilated mice undergo a lateral blunt thoracotomy via the 4th intercostal space. Following identification of the left anterior descending a ligature is passed underneath the vessel and both suture ends are threaded through an occluder. Then, both suture ends are passed through the chest wall, knotted to form a loop and left in the subcutaneous tissue. After chest closure and recovery for 5 days, mice are anesthetized again, chest skin is reopened and hanging weights are hooked up to the loop under ECG control.
At the end of the ischemia/reperfusion protocol, hearts can be stained with TTC for infarct size assessment or undergo perfusion fixation to allow morphometric studies in addition to histology and immunohistochemistry.

Watch this Scientific Journal Video about A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion at JoVE.com

A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion

Surgical trauma induces an inflammatory response. Cytokines and endogenous ligands are known to modulate myocardial infarct size following ischemia and reperfusion. We present a modified closed-chest model of murine ischemia and reperfusion using hanging weights to minimize effects of thoracotomy.

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ...

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