Objective: Investigating the cardiovascular system is challenging due to its complex regulation by humoral and neuronal factors. Despite this complexity, many existing research methods are limited to the assessment of a few parameters leading to an incomplete characterization of cardiovascular function. Thus, we aim to establish a murinein vivomodel for integrated assessment of the cardiovascular system under conditions of controlled heart rate. Utilizing this model, we assessed blood pressure, cardiac output, stroke volume, total peripheral resistance, and electrocardiogram (ECG).Hypothesis: We hypothesize that (i) ourin vivomodel can be utilized to investigate cardiac and vascular responses to pharmacological intervention with the α₁-agonist phenylephrine, and (ii) we can study cardiovascular function during artificial pacing of the heart, modulating cardiac function without a direct vascular effect.Methods: We included 12 mice that were randomly assigned to either vehicle or phenylephrine intervention through intraperitoneal administration. Mice were anesthetized with isoflurane and intubated endotracheally for mechanical ventilation. We measured blood pressureviaa solid-state catheter in the aortic arch, blood flowviaa probe on the ascending aorta, and ECG from needle electrodes on the extremities. Right atrium was electrically paced at a frequency ranging from 10 to 11.3 Hz before and after either vehicle or phenylephrine administration.Results: Phenylephrine significantly increased blood pressure, stroke volume, and total peripheral resistance compared to the vehicle group. Moreover, heart rate was significantly decreased following phenylephrine administration. Pacing significantly decreased stroke volume and cardiac output both prior to and after drug administration. However, phenylephrine-induced changes in blood pressure and total peripheral resistance were maintained with increasing pacing frequencies compared to the vehicle group. Total peripheral resistance was not significantly altered with increasing pacing frequencies suggesting that the effect of phenylephrine is primarily of vascular origin.Conclusion: In conclusion, thisin vivomurine model is capable of distinguishing between changes in peripheral vascular and cardiac functions. This study underlines the primary effect of phenylephrine on vascular function with secondary changes to cardiac function. Hence, thisin vivomodel is useful for the integrated assessment of the cardiovascular system.

Heterozygous mice (α₂^+/G301R mice) for the migraine-associated mutation (G301R) in the Na⁺,K⁺-ATPase α₂-isoform have decreased expression of cardiovascular α₂-isoform. The α₂^+/G301R mice exhibit a pro-contractile vascular phenotype associated with decreased left ventricular ejection fraction. However, the integrated functional cardiovascular consequences of this phenotype remain to be addressed in vivo. We hypothesized that the vascular response to α₂-isoform-specific inhibition of the Na⁺,K⁺-ATPase by ouabain is augmented in α₂^+/G301R mice leading to reduced cardiac efficiency. Thus, we aimed to assess the functional contribution of the α₂-isoform to in vivo cardiovascular function of wild-type (WT) and α₂^+/G301R mice. Blood pressure, stroke volume, heart rate, total peripheral resistance, arterial dP/dt, and systolic time intervals were assessed in anesthetized WT and α₂^+/G301R mice. To address rate-dependent cardiac changes, cardiovascular variables were compared before and after intraperitoneal injection of ouabain (1.5 mg/kg) or vehicle during atrial pacing. The α₂^+/G301R mice showed an enhanced ouabain-induced increase in total peripheral resistance associated with reduced efficiency of systolic development compared to WT. When the hearts were paced, ouabain reduced stroke volume in α₂^+/G301R mice. In conclusion, the ouabain-induced vascular response was augmented in α₂^+/G301R mice with consequent suppression of cardiac function.

Two α-isoforms of the Na⁺,K⁺-ATPase (α₁ and α₂) are expressed in the cardiovascular system, and it is unclear which isoform is the preferential regulator of contractility. Mice heterozygous for the familial hemiplegic migraine type 2 (FHM2) associated mutation in the α₂-isoform (G301R; α₂^+/G301R mice) have decreased expression of cardiac α₂-isoform but elevated expression of the α₁-isoform. We aimed to investigate the contribution of the α₂-isoform function to the cardiac phenotype of α₂^+/G301R hearts. We hypothesized that α₂^+/G301R hearts exhibit greater contractility due to reduced expression of cardiac α₂-isoform. Variables for contractility and relaxation of isolated hearts were assessed in the Langendorff system without and in the presence of ouabain (1 µM). Atrial pacing was performed to investigate rate-dependent changes. The α₂^+/G301R hearts displayed greater contractility than WT hearts during sinus rhythm, which was rate-dependent. The inotropic effect of ouabain was more augmented in α₂^+/G301R hearts than in WT hearts during sinus rhythm and atrial pacing. In conclusion, cardiac contractility was greater in α₂^+/G301R hearts than in WT hearts under resting conditions. The inotropic effect of ouabain was rate-independent and enhanced in α₂^+/G301R hearts, which was associated with increased systolic work.