This work is supported by the American Heart Association 14FTF20370058 (DMA) and NIH T32 HL007101-35 (DMA).

1. Conductance Catheter Preparations and Pressure Calibration
<ol>
	<li>Connect conductance catheter to the hemodynamic catheter module. Electronically calibrate pressure and volume measurements by recording preset pressure and volume set on the catheter module. Record a tracing of 0 mm Hg and 25 mm Hg (Figure 1A) and assign voltages to both pressure tracings (Figure&#160;1B and 1C). Similarly, record a volume tracing of 5 RVU and 25 RVU (Figure 1D) and assign voltages...

<ol>
	<li>Clark, J. E., Marber, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advancements+in+pressure-volume+catheter+technology+-+stress+remodelling+after+infarction.">Advancements in pressure-volume catheter technology - stress remodelling after infarction.</a> Exp Physiol. 98 (3), 614-621 (2013).</li><li>Sunagawa, K., Maughan, W. L., Burkhoff, D., Sagawa, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Left+ventricular+interaction+with+arterial+load+studied+in+isolated+canine+ventricle.">Left ventricular interaction with arterial load studied in isolated canine ventricle.</a> Am J Physiol. 245 (Pt 1), H773-H780 (1983).</li><li>Suga, H., Sagawa, K., Demer, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determinants+of+instantaneous+pressure+in+canine+left+ventricle.+Time+and+volume+specification.">Determinants of instantaneous pressure in canine left ventricle. Time and volume specification.</a> Circ Res. 46 (2), 256-263 (1980).</li><li>Suga, H., Sagawa, K., Shoukas, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Load+independence+of+the+instantaneous+pressure-volume+ratio+of+the+canine+left+ventricle+and+effects+of+epinephrine+and+heart+rate+on+the+ratio.">Load independence of the instantaneous pressure-volume ratio of the canine left ventricle and effects of epinephrine and heart rate on the ratio.</a> Circ Res. 32 (3), 314-322 (1973).</li><li>Kass, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+left+ventricular+mechanics+from+acute+VDD+pacing+in+patients+with+dilated+cardiomyopathy+and+ventricular+conduction+delay.">Improved left ventricular mechanics from acute VDD pacing in patients with dilated cardiomyopathy and ventricular conduction delay.</a> Circulation. 99 (12), 1567-1573 (1999).</li><li>Kass, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diastolic+Compliance+of+Hypertrophied+Ventricle+Is+Not+Acutely+Altered+by+Pharmacological+Agents+Influencing+Active+Processes.">Diastolic Compliance of Hypertrophied Ventricle Is Not Acutely Altered by Pharmacological Agents Influencing Active Processes.</a> Annals of Internal Medicine. 119 (6), 466-473 (1993).</li><li>Georgakopoulos, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+murine+left+ventricular+pressure-volume+relations+by+miniaturized+conductance+micromanometry.">In vivo murine left ventricular pressure-volume relations by miniaturized conductance micromanometry.</a> Am J Physiol. 274 (4 pt 2), H1416-H1422 (1998).</li><li>Cingolani, O. H., Kass, D. 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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anaesthesia+of+small+rodents+during+magnetic+resonance+imaging.">Anaesthesia of small rodents during magnetic resonance imaging.</a> Methods. 43 (1), 68-78 (2007).</li><li>Georgakopoulos, D., Kass, D. 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S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=beta-Arrestin-biased+AT1R+stimulation+promotes+cell+survival+during+acute+cardiac+injury.">beta-Arrestin-biased AT1R stimulation promotes cell survival during acute cardiac injury.</a> Am J Physiol Heart Circ Physiol. 303 (8), H1001-H1010 (2012).</li><li>Suga, H., Sagawa, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mathematical+Interrelationship+between+Instantaneous+Ventricular+Pressure-Volume+Ratio+and+Myocardial+Force-Velocity+Relation.">Mathematical Interrelationship between Instantaneous Ventricular Pressure-Volume Ratio and Myocardial Force-Velocity Relation.</a> Annals of Biomedical Engineering. 1 (2), 160-181 (1972).</li><li>Suga, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ventricular+energetics.">Ventricular energetics.</a> Physiol Rev. 70 (2), 247-277 (1990).</li><li>Kass, D. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+contractile+state+on+curvilinearity+of+in+situ+end-systolic+pressure-volume+relations.">Influence of contractile state on curvilinearity of in situ end-systolic pressure-volume relations.</a> Circulation. 79 (1), 167-178 (1989).</li><li>Little, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+left+ventricular+dP/dtmax-end-diastolic+volume+relation+in+closed-chest+dogs.">The left ventricular dP/dtmax-end-diastolic volume relation in closed-chest dogs.</a> Circ Res. 56 (6), 808-815 (1985).</li><li>Glower, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Linearity+of+the+Frank-Starling+relationship+in+the+intact+heart:+the+concept+of+preload+recruitable+stroke+work.">Linearity of the Frank-Starling relationship in the intact heart: the concept of preload recruitable stroke work.</a> Circulation. 71 (5), 994-1009 (1985).</li><li>Sharir, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ventricular+systolic+assessment+in+patients+with+dilated+cardiomyopathy+by+preload-adjusted+maximal+power.+Validation+and+noninvasive+application.">Ventricular systolic assessment in patients with dilated cardiomyopathy by preload-adjusted maximal power. Validation and noninvasive application.</a> Circulation. 89 (5), 2045-2053 (1994).</li><li>Baan, J., Van der Velde, E. 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The authors have nothing to disclose.

We describe a method for perfoming pressure-volume loop analysis using a conductance catheter in mice, to derive comprehensive analyses of both cardiac contractility and relaxation. Suga, Sagawa and colleagues utilized pressure-volume loops to define measures of cardiac contractility, specifically the slope of the ESPVR, or the end-systolic elastance (Ees), and Emax. Elastance, defined by the ratio of pressure to volume (P/V), varies over the duration of systole. During each systole, the instantaneo...

Cardiac pressure volume loop analysis provides detailed information of cardiac function and are the gold standard for functional assessment 1. While imaging techniques such as echocardiography or cardiac MRI provide functional measures, these measures are highly dependent on loading conditions. Load-independent measures of cardiac contractility and relaxation require dynamic measurements of the ventricular pressure and volume relation over a range of preload and afterload. This understanding of the pressure-volume relation arises from the groundbreaking work of Sagawa and colleagues 2,3. They demonstrated in ex vivo perfused canine hearts that the pressure-volume loop derived contractility measures were independent of loading conditions 4.
In vivo application of these analyses became possible with the development of conductance catheters in the 1980s. This technical advance allowed Kass and colleagues to perform pressure-volume loop analysis in humans 5,6. Miniaturization of conductance catheters and improvements in surgical techniques in the late 1990&#8217;s 7 made analysis of rodent cardiac function feasible, allowing for genetic and pharmacologic studies to be performed. This advance has since lead to the widespread use of pressure-volume loop analysis and has generated a great deal of insight into mammalian cardiac physiology.
A key concept in the use of conductance catheters and the interpretation of data obtained from it is the relationship between volume and conductance. Conductance is inversely related to voltage, which is measured using a catheter with electrodes placed proximally, usually placed below the aortic valve, and distally, at the LV apex 8. Changes in voltage or conductance are measured by changes in current flowing from proximal to distal electrode. Although the blood pool contributes significantly to conductance, the contribution of the ventricular wall, termed parallel conductance (Vp), to measured conductance must be subtracted to obtain absolute LV volume measurements.
The methods to perform this correction, called a saline calibration, are discussed in the protocol below. The mathematical relationship between conductance and volume, described by Baan and colleagues, is that volume=1/&#945;; (&#961;L2)(G-Gp), where &#945;=uniform field correction factor, &#961;=blood resistivity, L=distance between the electrodes, G=conductance and Gp=non-blood conductance 9. Of note, the uniform field correction factor in mice approaches 1.0 due to small chamber volumes10. Coupled with pressure transducers, the conductance catheter provides real time simultaneous pressure and volume data.
Cardiac pressure-volume analysis presents particular advantages over other measures of cardiac function, as they allow for measurement of ventricular function independent of loading conditions and of heart rate. Specific load-independent cardiac indices of contractility include: end-systolic pressure volume relation (ESPVR), dP/dtmax&#8211;end-diastolic volume relation, maximal elastance (Emax) and preload recruitable stroke work (PRSW). A load-independent measure of diastolic function is the end-diastolic pressure volume relationship (EDPVR) 11. The following protocol describes the conduct of cardiac pressure volume loop analysis, using both a carotid and an apical approach. While the methodology to perform these studies have been described in detail previously 8,11, we will review key steps to obtain precise pressure-volume measurements, including both saline and cuvette calibration correction, and provide a visual demonstration of these procedures. &#160;&#160;Research with animals carried out for this study was handled according to approved protocols and animal welfare regulations of Duke University Medical Center&#8217;s Institutional Animal Care and Use Committee.

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			<td>AnaSed (xylazine)&#160;</td>
			<td>Lloyd Laboratories</td>
			<td>NADA no. 139-236</td>
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			<td>VIP3000</td>
			<td>Matrx Medical Inc.</td>
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			<td>Anesthesia machine</td>
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			<td>Harvard Apparatus</td>
			<td>Model 683</td>
			<td>Surgical Equipment</td>
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			<td>Tubing kit</td>
			<td>Harvard Apparatus</td>
			<td>72-1049</td>
			<td>Surgical Equipment</td>
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			<td>Homeothermic Blanket&#160;</td>
			<td>Kaz Inc.</td>
			<td>5628</td>
			<td>Surgical Equipment</td>
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			<td>Stereo microscope</td>
			<td>Carl Zeiss Optical Inc.</td>
			<td>Stemi 2000</td>
			<td>Surgical Equipment</td>
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			<td>Illuminator</td>
			<td>Cole&#8211;Parmer</td>
			<td>41720</td>
			<td>Surgical Equipment</td>
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			<td>Dumont no. 55 Dumostar Forceps&#160;</td>
			<td>Fine Science Tools Inc</td>
			<td>11295-51</td>
			<td>Surgical Instruments</td>
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			<td>Graefe forceps, curved&#160;</td>
			<td>Fine Science Tools Inc</td>
			<td>11052-10</td>
			<td>Surgical Instruments</td>
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			<td>Moria MC31 forceps&#160;</td>
			<td>Fine Science Tools Inc</td>
			<td>11370-31</td>
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			<td>Fine Science Tools Inc</td>
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			<td>Surgical Instruments</td>
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			<td>Iris scissors&#160;</td>
			<td>Fine Science Tools Inc</td>
			<td>14041-10</td>
			<td>Surgical Instruments</td>
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			<td>Halsey needle holder&#160;</td>
			<td>Fine Science Tools Inc</td>
			<td>12501-13</td>
			<td>Surgical Instruments</td>
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			<td>Olsen&#8211;Hegar needle holder&#160;</td>
			<td>Fine Science Tools Inc</td>
			<td>12002-12</td>
			<td>Surgical Instruments</td>
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			<td>spring scissors</td>
			<td>Fine Science Tools Inc</td>
			<td>15610-08</td>
			<td>Surgical Instruments</td>
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		<tr>
			<td>disposable underpads</td>
			<td>Kendall/Tyco Healthcare</td>
			<td>1038</td>
			<td>Surgical Supplies</td>
		</tr>
		<tr>
			<td>Sterile gauze sponges, sterile&#160;</td>
			<td>Dukal</td>
			<td>62208</td>
			<td>Surgical Supplies</td>
		</tr>
		<tr>
			<td>Cotton-tipped applicators, sterile&#160;</td>
			<td>Solon</td>
			<td>368</td>
			<td>Surgical Supplies</td>
		</tr>
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			<td>Surgical suture,&#160; silk, 6-0&#160;</td>
			<td>DemeTECH</td>
			<td>FT-639-1</td>
			<td>Surgical Supplies</td>
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			<td>1 cc Insulin syringes&#160;</td>
			<td>Becton Dickenson</td>
			<td>329412</td>
			<td>Surgical Supplies</td>
		</tr>
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			<td>Access-9&#8482; Hemostasis Valve</td>
			<td>Merit Medical&#160;</td>
			<td>MAP111</td>
			<td>Hemodynamic equipment</td>
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		<tr>
			<td>Sphygmomanometer</td>
			<td>Baumanometer</td>
			<td>320</td>
			<td>Hemodynamic equipment</td>
		</tr>
		<tr>
			<td>Millar PV system MPVS-300/400 or MPVS Ultra (includes calibration cuvette)</td>
			<td>ADInstruments Inc</td>
			<td></td>
			<td>Hemodynamic equipment</td>
		</tr>
		<tr>
			<td>1.4F conductance catheter&#160;</td>
			<td>ADInstruments Inc</td>
			<td>SPR-839</td>
			<td>Hemodynamic equipment</td>
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			<td>PowerLab 4/30 with Chart Pro</td>
			<td>ADInstruments Inc.</td>
			<td>ML866/P</td>
			<td>Hemodynamic software</td>
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			<td>animal clipper</td>
			<td>Wahl</td>
			<td>8787-450A</td>
			<td>Miscellaneous</td>
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			<td>Intradermic tubing PE-10 (Becton Dickinson, cat. no. )</td>
			<td>Becton Dickenson</td>
			<td>427401</td>
			<td>Miscellaneous</td>
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			<td>Intradermic tubing PE-50 (Becton Dickinson, cat. no.)</td>
			<td>Becton Dickenson</td>
			<td>427411</td>
			<td>Miscellaneous</td>
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			<td>Needle assortment (18, 25 and 30 gauge; Thomas Scientific)</td>
			<td></td>
			<td></td>
			<td>Miscellaneous</td>
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			<td>0.9% (wt/vol) sodium chloride injection, USP , cat. no. )</td>
			<td>Hospira</td>
			<td>NDC no. 0409-4888-50</td>
			<td>Miscellaneous</td>
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		<tr>
			<td>Surgical tape</td>
			<td></td>
			<td></td>
			<td>Miscellaneous</td>
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		<tr>
			<td>Alconox (Alconox Inc.) for catheter cleaning</td>
			<td>ADInstruments Inc.</td>
			<td></td>
			<td>Miscellaneous</td>
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cardiac pressure-volume loop analysis using conductance catheters in mice

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular systolic and diastolic function. Measures of ventricular contractility and compliance are obtained through examination of cardiac response to changes in afterload and preload. These techniques were originally developed nearly three decades ago to measure cardiac function in large mammals and humans. The application of these analyses to small mammals, such as mice, has been accomplished through the optimization of microsurgical techniques and creation of conductance catheters. Conductance catheters allow for estimation of the blood pool by exploiting the relationship between electrical conductance and volume. When properly performed, these techniques allow for testing of cardiac function in genetic mutant mouse models or in drug treatment studies. The accuracy and precision of these studies are dependent on careful attention to the calibration of instruments, systematic conduct of hemodynamic measurements and data analyses. We will review the methods of conducting pressure-volume loop experiments using a conductance catheter in mice.

Pressure-volume loop analysis can be used to measure cardiac function in genetically modified mice 14,15 or mice undergoing drug studies16. Representative pressure volume loops are provided from previously published work 16 investigating the effect of &#223;-Arrestin biased AT1R ligand, TRV120023. To test whether TRV120023 affects cardiac function in vivo, pressure-volume loop analysis was performed on wild type mice receiving conventional and novel angiotensin receptor blockers....

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Cardiac Pressure-Volume Loop Analysis Using Conductance Catheters in Mice

Cardiac pressure-volume loop analysis is the most comprehensive way to measure cardiac function in the intact heart. We describe a technique to perform and analyze cardiac pressure volume loops, using conductance catheters.

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular ...

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19.2K visualizações.  Duke University Medical Center. Cardiac pressure-volume loop analysis is the most comprehensive way to measure cardiac function in the intact heart. We describe a technique to perform and analyze cardiac pressure volume loops, using conductance catheters. 