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Medicine

Cardiac Stress Test Induced by Dobutamine and Monitored by Cardiac Catheterization in Mice

Published: February 10th, 2013

DOI:

10.3791/50050

1Instituto de Ciencias, Facultad de Medicina, Clínica Alemana Universidad del Desarrollo

We describe the protocol to perform a cardiac stress test induced by dobutamine and monitored by cardiac catheterization in normal mice. Also we show its application to unmask subclinical cardiac disease in high fat diet-induced obese mice.

Dobutamine is a β-adrenergic agonist with an affinity higher for receptor expressed in the heart (β1) than for receptors expressed in the arteries (β2). When systemically administered, it increases cardiac demand. Thus, dobutamine unmasks abnormal rhythm or ischemic areas potentially at risk of infarction.

Monitoring of heart function during a cardiac stress test can be performed by either ecocardiography or cardiac catheterization. The latter is an invasive but more accurate and informative technique that the former.

Cardiac stress test induced by dobutamine and monitored by cardiac catheterization accomplished as described here allows, in a single experiment, the measurement of the following hemodynamic parameters: heart rate (HR), systolic pressure, diastolic pressure, end-diastolic pressure, maximal positive pressure development (dP/dtmax) and maximal negative pressure development (dP/dtmin), at baseline conditions and under increasing doses of dobutamine.

As expected, in normal mice we observed a dobutamine dose-related increase in HR, dP/dtmax and dP/dtmin. Moreover, at the highest dose tested (12 ng/g/min) the cardiac decompensation of high fat diet-induced obese mice was unmasked.

Protocol was approved by the Ethic Committee of Facultad de Medicina Clinica Alemana-Universidad del Desarrollo.

I. Preparing Dobutamine Infusion

  1. Dissolve 10 mg of dobutamine in 20 ml of sterile distillated water, in order to obtain a stock solution of 500 μg/ml dobutamine. Aliquot and store at -20 °C. This solution can be used at least for 3 months.
  2. Thaw an aliquot of dobutamine stock solution at room temperature.
  3. Dilute dobutamine stock solution.......

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The arterial pressure signal is defined by systolic and diastolic pressure. When the pressure sensor is inside the left ventricle, its pressure (LVP) waveform is characterized by a drop to zero of the diastolic pressure and the appearance of the left atrial contraction before ventricle contraction (Figure 1). At baseline condition, ketamine-xylazine anesthetized normal mice had HR of 280 ± 24, Pmax of 107 ± 8, Pmin of 5 ± 1, EDP of 14 ± 2, dP/dtmax

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Cardiac stress test induced by dobutamine and monitored by cardiac catheterization is laborious. Nonetheless, following the protocol here describe and with a short time of training, it is possible to assess six hemodynamic parameters in a single experiment that last approximately one hour.

The critical steps of the protocol here presented are the cannulations of blood vessels. Regarding the cannulation of carotid artery, the incision performed should be deep enough to break the three tissue la.......

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We thank Dr. Helio Salgado, Renata Lataro and Mauro de Oliveira, School of Medicine of Ribeirão Preto, University of Sao Paulo and Dr. Ben Janssen, Cardiovascular Research Institute Maastricht, Maastricht University, for generous assistance during the set up process.

This work was supported by FONDECYT grant N° 11090114 to S.D.C.

....

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Name Company Catalog Number Comments
Reagents
PE-50/10 Warner Instruments 64-0752
Silk thread 6/0 HR17 Tagum SN0713K
Xylacin 20 mg/ml Laboratorio Centrovet
Ketamine 100 mg/ml Drag Pharma
Sodium chloride 0.9% Lab Sanderson S.A.
Dobutamine hydrochloride Sigma-Aldrich D0676
Syringe U-100 Insulin 29G x ½" Terumo Medical Co.
Forceps Dissecting Micro 11.5 cm Style 7 Lawton Medizintechnik 09-0959
Graefe Forceps Cvd 0.7mm 7cm Lawton Medizintechnik 62-0263
Clamps Dieffenbach bulldog Cl Str 38 mm Lawton Medizintechnik 60-010
Vannas Scissors 8 cm Str Fh Lawton Medizintechnik 63-1400
Equipment
SPR-671 MiKro-Tip Pressure catheter Millar instruments 840-6719
PCU-2000 Pressure Control Unit Millar instruments 880-0129
PowerLab 4/30 ADinstruments Pty Ltd. ML866
LabChartPro 7 ADinstruments Pty Ltd. MLU260/7
Legato200 Infusion Pump KdScientific KD-KDS210P
TCAT-2LV Temperature controller and isothermal heating plate PhysiTemp instruments Inc.
Medical Oxygen supply Indura
Rectal probe ADinstruments Pty Ltd. MLT1404
Trinocular microscope, axial illumination LW Scientific Z2B-TRI-ETNE, ILP-1502-LTS1, ILP-1502-DGGF

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