A subscription to JoVE is required to view this content. Sign in or start your free trial.
* These authors contributed equally
Here we describe a cardiac pressure-volume loop analysis under increasing doses of intravenously infused isoproterenol to determine the intrinsic cardiac function and the β-adrenergic reserve in mice. We use a modified open-chest approach for the pressure-volume loop measurements, in which we include ventilation with positive end-expiratory pressure.
Determination of the cardiac function is a robust endpoint analysis in animal models of cardiovascular diseases in order to characterize effects of specific treatments on the heart. Due to the feasibility of genetic manipulations the mouse has become the most common mammalian animal model to study cardiac function and to search for new potential therapeutic targets. Here we describe a protocol to determine cardiac function in vivo using pressure-volume loop measurements and analysis during basal conditions and under β-adrenergic stimulation by intravenous infusion of increasing concentrations of isoproterenol. We provide a refined protocol including ventilation support taking into account the positive end-expiratory pressure to ameliorate negative effects during open-chest measurements, and potent analgesia (Buprenorphine) to avoid uncontrollable myocardial stress evoked by pain during the procedure. All together the detailed description of the procedure and discussion about possible pitfalls enables highly standardized and reproducible pressure-volume loop analysis, reducing the exclusion of animals from the experimental cohort by preventing possible methodological bias.
Cardiovascular diseases typically affect cardiac function. This issue points out the importance in assessing in vivo detailed cardiac function in animal disease models. Animal experimentation is surrounded by a frame of the three Rs (3Rs) guiding principles (Reduce/Refine/Replace). In case of understanding complex pathologies involving systemic responses (i.e., cardiovascular diseases) at the current developmental level, the main option is to refine the available methods. Refining will also lead to a reduction of the required animal numbers due to less variability, which improves the power of the analysis and conclusions. In addition, combination of cardiac contractil....
All animal experiments were approved and performed according to the regulations of the Regional Council of Karlsruhe and the University of Heidelberg (AZ 35-9185.82/A-2/15, AZ 35-9185.82/A-18/15, AZ 35-9185.81/G131/15, AZ 35-9185.81/G121/17) conform to the guidelines from Directive 2010/63/EU of the European Parliament on the protection of animals used for scientific purposes. Data shown in this protocol are derived from wild type C57Bl6/N male mice (17 ± 1.4 weeks of age). Mice were maintained under specified .......
The pressure volume-loop (PVL) measurement is a powerful tool to analyze cardiac pharmacodynamics of drugs and to investigate the cardiac phenotype of genetically modified mouse models under normal and pathological conditions. The protocol allows the assessment of cardiac β-adrenergic reserve in the adult mouse model. Here we describe an open-chest method under isoflurane anesthesia combined with buprenorphine (analgesic) and pancuronium (muscle relaxant), which focuses on the cardiac response to β-adrenergic s.......
Here, we provide a protocol to analyze the in vivo cardiac function in mice under increasing β-adrenergic stimulation. The procedure can be used to address both, baseline parameters of cardiac function and the adrenergic reserve (e.g., inotropy and chronotropy) in genetically modified mice or upon interventions. The most prominent advantage of pressure-volume loop (PVL) measurements as compared to other means of determining cardiac function is the analysis of intrinsic, load-independent cardiac function. All other m.......
We are thankful to Manuela Ritzal, Hans-Peter Gensheimer, Christin Richter and the team from the Interfakultäre Biomedizinische Forschungseinrichtung (IBF) from the Heidelberg University for expert technical assistance.
This work was supported by the DZHK (German Centre for Cardiovascular Research), the BMBF (German Ministry of Education and Research), a Baden-Württemberg federal state Innovation fonds and the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) Project-ID 239283807 - TRR 152, FOR 2289 and the Collaborative Research Center (SFB) 1118.
....Name | Company | Catalog Number | Comments |
1.4F SPR-839 catheter | Millar Instruments, USA | 840-8111 | |
1 ml syringes | Beckton Dickinson, USA | REF303172 | |
Bio Amplifier | ADInstruments, USA | FE231 | |
Bridge-Amplifier | ADInstruments, USA | FE221 | |
Bovine Serum Albumin | Roth, Germany | 8076.2 | |
Buprenorphine hydrochloride | Bayer, Germany | 4007221026402 | |
Calibration cuvette | Millar, USA | 910-1049 | |
Differential pressure transducer MPX | Hugo Sachs Elektronik- Harvard Apparatus, Germany | Type 39912 | |
Dumont Forceps #5/45 | Fine Science tools Inc. | 11251-35 | |
Dumont Forceps #7B | Fine Science tools Inc. | 11270-20 | |
Graefe Forceps | Fine Science tools Inc. | 11051-10 | |
GraphPad Prism | GraphPad Software | Ver. 8.3.0 | |
EcoLab-PE-Micotube | Smiths, USA | 004/310/168-1 | |
Etomidate Lipuro | Braun, Germany | 2064006 | |
Excel | Microsoft | ||
Heparin | Ratiopharm, Germany | R26881 | |
Hot plate and control unit | Labotec, Germany | Hot Plate 062 | |
Isofluran | Baxter, Germany | HDG9623 | |
Isofluran Vaporizer | Abbot | Vapor 19.3 | |
Isoprenalinhydrochloride | Sigma-Aldrich, USA | I5627 | |
Fine Bore Polythene tubing 0.61 mm OD, 0.28 mm ID | Smiths Medical International Ltd, UK | Ref. 800/100/100 | |
MiniVent ventilator for mice | Hugo Sachs Elektronik- Harvard Apparatus, Germany | Type 845 | |
MPVS Ultra PVL System | Millar Instruments, USA | ||
NaCl | AppliChem, Germany | A3597 | |
NaCl 0.9% isotonic | Braun, Germany | 2350748 | |
Pancuronium-bromide | Sigma-Aldrich, USA | BCBQ8230V | |
Perfusor 11 Plus | Harvard Apparatus | Nr. 70-2209 | |
Powerlab 4/35 control unit | ADInstruments, USA | PL3504 | |
Rechargeable cautery-Set | Faromed, Germany | 09-605 | |
Scissors | Fine Science tools Inc. | 140094-11 | |
Software LabChart 7 Pro | ADInstruments, USA | LabChart 7.3 Pro | |
Standard mouse food | LASvendi GmbH, Germany | Rod18 | |
Stereo microscope | Zeiss, Germany | Stemi 508 | |
Surgical suture 8/0 | Suprama, Germany | Ch.B.03120X | |
Venipuncture-cannula Venflon Pro Safty 20-gauge | Beckton Dickinson, USA | 393224 | |
Vessel Cannulation Forceps | Fine Science tools Inc. | 00574-11 | |
Water bath | Thermo Fisher Scientific, USA | ||
Syringe filter (Filtropur S 0.45) | Sarstedt, Germany | Ref. 83.1826 |
This article has been published
Video Coming Soon
ABOUT JoVE
Copyright © 2024 MyJoVE Corporation. All rights reserved