Indiana University-Purdue University Indianapolis (IUPUI) / Autonomic innervation of the heart begins in utero and continues during the neonatal phase of life. A balance between the sympathetic and parasympathetic arms of the autonomic nervous system is required to regulate heart rate as well as the force of each contraction. Our lab studies the development of sympathetic innervation of the early postnatal heart in a conditional knockout (cKO) of Src homology protein tyrosine phosphatase 2 (Shp2). Shp2 is a ubiquitously expressed non-receptor phosphatase involved in a variety of cellular functions including survival, proliferation, and differentiation. We targeted Shp2 in post-migratory neural crest (NC) lineages using our novel Periostin-Cre. This resulted in a fully penetrant mouse model of diminished cardiac sympathetic innervation and concomitant bradycardia that progressively worsen. Shp2 is thought to mediate its basic cellular functions through a plethora of signaling cascades including extracellular signal-regulated kinases (ERK) 1 and 2. We hypothesize that abrogation of downstream ERK1/2 signaling in NC lineages is primarily responsible for the failed sympathetic innervation phenotype observed in our mouse model. Shp2 cKOs are indistinguishable from control littermates at birth and exhibit no gross structural cardiac anomalies; however, in vivo electrocardiogram (ECG) characterization revealed sinus bradycardia that develops as the Shp2 cKO ages. Significantly, 100% of Shp2 cKOs die within 3 weeks after birth. Characterization of the expression pattern of the sympathetic nerve marker tyrosine hydroxylase (TH) revealed a loss of functional sympathetic ganglionic neurons and reduction of cardiac sympathetic axon density in Shp2 cKOs. Shp2 cKOs exhibit lineage-specific suppression of activated pERK1/2 signaling, but not of other downstream targets of Shp2 such as pAKT (phosphorylated-Protein kinase B). Interestingly, restoration of pERK signaling via lineage-specific expression of constitutively active MEK1 (Mitogen-activated protein kinase kinase1) rescued TH-positive cardiac innervation as well as heart rate. These data suggest that the diminished sympathetic cardiac innervation and the resulting ECG abnormalities are a result of decreased pERK signaling in post-migratory NC lineages.

INTRODUÇÃO: O treinamento físico provoca adaptações cardiovasculares, sendo que maiores adaptações estruturais cardíacas são observadas em atletas de elite, que realizam treinamento físico de alta intensidade com o objetivo de aumento no rendimento. Além disto, atletas apresentam diminuição da frequência cardíaca de repouso, embora os mecanismos que explicam a bradicardia em atletas ainda são controversos. O nível das adaptações pode variar e fatores como o tipo de modalidade esportiva, tempo e nível de treinamento físico podem contribuir para tal variação. Desta forma, o objetivo deste estudo foi investigar a influência de modalidades esportivas (ciclismo, corrida de longa distância e remo) e da periodização do treinamento físico nas adaptações estruturais e funcionais, autonômicas e não autonômicas cardíacas e vasculares em atletas de elite no repouso, na inclinação ortostática e nas 24 horas. MÉTODOS: Neste estudo experimental longitudinal prospectivo foram avaliados 13 ciclistas, 13 corredores e 11 remadores de elite, saudáveis (entre 20 e 36 anos; masculino), engajados em treinamento físico competitivo, em 2 períodos de treinamento: período básico-PB e período competitivo-PC. Avaliação da capacidade funcional máxima foi feita por teste cardiorrespiratório. Adaptações estruturais cardíacas foram avaliadas por meio do ecocardiograma bidimensional com doppler. Frequência cardíaca intrínseca foi estudada por meio do duplo bloqueio farmacológico (atropina 0,04 mg/kg e esmolol 500 g/kg, i.v.). Frequência cardíaca e pressão arterial foram registradas continuamente no repouso e no teste de inclinação ortostática por meio de ECG e monitor de pressão arterial, respectivamente (500Hz). A variabilidade da frequência cardíaca e pressão arterial foram analisadas pelo método auto-regressivo. Frequência cardíaca e pressão arterial de 24 horas foram aquisitadas pelo Holter e Mapa, respectivamente. Avaliação da sensibilidade barorreflexa espontânea foi calculada pelo método da sequência. Os dados foram apresentados como mediana e variação interquartil. RESULTADOS: Remadores apresentaram maior VO2max que os corredores no PC (p<0,04). Ciclistas e remadores apresentaram maior VO2max no PC comparado ao PB (p<0,05). Corredores apresentaram maiores diâmetros diastólicos do ventrículo esquerdo que ciclistas (p<0,06) e remadores (p<0,01) no PB. Corredores e ciclistas apresentaram maiores diâmetros diastólicos do ventrículo esquerdo que remadores (p<0,004) no PC. Corredores apresentaram maiores índices de massa do ventrículo esquerdo que ciclistas (p<0,04) no PB e ciclistas maiores que remadores (p<0,03) no PC. Ciclistas foram os únicos atletas que apresentaram a reversão do remodelamento cardíaco no PB (p<0,04). No repouso, corredores mostraram menor frequência cardíaca que ciclistas no PC ciclistas = 50(45/55), corredores = 44(43/47), remadores = 44(43/53)bpm, p<0,03]. Corredores e remadores mostraram maior efeito vagal [ciclistas = 41(36/46), corredores = 55(48/59), remadores = 50(42/66)bpm, p=0,03] e maior frequência cardíaca intrínseca ciclistas = 84(82/87), corredores = 92(87/94), remadores = 96(85/101)bpm, p=0,03] que ciclistas no PC. Remadores tinham maior frequência cardíaca intrínseca que ciclistas no PB ciclistas = 88(86/92), corredores = 91(82/99), remadores = 95(90/101)bpm, p=0,03]. PC comparado ao PB diminuiu o efeito vagal 41(36/46) vs. 48(43/51)bpm, p<0,05] e frequência cardíaca intrínseca 84(82/87) vs. 88(86/92), p<0,05] dos ciclistas. No teste de inclinação, corredores mostraram menor aumento da frequência cardíaca que ciclistas no PB ciclistas = 64(49/73), corredores = 46(43/55), remadores = 53(42/77)%, p<0,05]. Nem a modalidade ou período de treinamento físico influenciaram os índices de variabilidade da frequência cardíaca no repouso ou na inclinação ortostática. A modalidade esportiva e o período de treinamento físico influenciaram a pressão arterial e sua variabilidade, no repouso e na inclinação ortostática. Remadores apresentaram menor sensibilidade barorreflexa espontânea que corredores no PC (p=0,03). Tanto a modalidade quanto o período de treinamento físico influenciaram a resposta da frequência cardíaca de 24 horas, sua variabilidade e a resposta da pressão arterial de 24 horas. CONCLUSÃO: Tanto a modalidade esportiva quanto o período de treinamento físico influenciaram nas adaptações estruturais cardíacas, intrínsecas e autonômicas cardíacas e vasculares. Entretanto a modalidade esportiva parece influenciar mais expressivamente essas adaptações / INTRODUCTION: Physical exercise training provokes cardiovascular adaptations and the highest structural cardiac adaptations are observed in elite athletes who perform high intensity training with the objective of increasing their physical performance. Besides, athlete shows decrease on resting heart rate. However, the mechanisms that explain the bradycardia in athletes are still controversial. The level of these adaptations may vary and some factors as the type of sport modality, time and level of physical training can contribute to such different responses. Thus, the aim of this study was to investigate the influence of sport modalities (cycling, long distane runner and rower) and physical training periodization on cardiac structural and functional, cardiac autonomic and non-autonomic and vascular adaptations in elite athletes at rest, tilt table test and within 24 hours. METHODS: In this prospective longitudinal experimental study, 13 cyclists, 13 runners and 11 rowers, healthy (20 to 36 years old; male), engaged in competitive training were evaluated in 2 periods of training: basic period BP and competitive period -CP. Maximal functional capacity was evaluated by cardiopulmonary test. Cardiac structural adaptations were evaluated by two-dimensional echocardiography with doppler. Intrinsic heart rate was studied by means of double pharmacological blockade (atropine 0.04 mg/kg and esmolol 500 g/kg, iv.). Heart rate and blood pressure were recorded continuously at rest and tilt table test by means of ECG and arterial blood pressure monitor, respectively (500Hz). The heart rate and blood pressure variabilities were analyzed by autoregressive method. Heart rate and blood pressure within 24 hours were recorded using Holter and blood pressure ambulatory monitor, respectivamente. Spontaneous baroreflex sensitivity was calculated using the sequence method. The data were presented as median and interquartile range. RESULTS: Rowers showed higher VO2max than runners at CP (p<0.04). Cyclists and rowers showed higher VO2max at CP compared to BP (p<0.05). Runners presented higher left ventricular diastolic diameters than cyclists (p<0.06) and rowers (p<0.01) at BP. Runners and cyclists presented higher left ventricular diastolic diameters than rowers (p<0.004) at CP. Runners showed higher left ventricular mass index than cyclistas (p<0.04) at BP and cyclists higher than rowers (p<0.03) at CP. Cyclists were the only athletes who had a reversal of cardiac remodeling at BP (p<0.04). At rest, runners showed lower heart rate than cyclists at CP cyclists = 50(45/55), runners = 44(43/47), rowers = 44(43/53)bpm, p<0.03]. Runners and rowers showed higher vagal effect [cyclists = 41(36/46), runners = 55(48/59), rowers = 50(42/66)bpm, p=0.03] and higher intrinsic heart rate cyclists = 84(82/87), runners = 92(87/94), rowers = 96(85/101)bpm, p=0,03] than cyclists at CP. Rowers had higher intrinsic herat rate than cyclists at BP cyclists = 88(86/92), runners = 91(82/99), rowers = 95(90/101)bpm, p=0.03]. CP compared with BP decreased the vagal effect 41(36/46) vs. 48(43/51)bpm, p<0.05] and intrinsic heart rate 84(82/87) vs. 88(86/92), p<0.05] of cyclists. At tilt table test, runners showed smaller increase in heart rate than cyclists at BP cyclists = 64(49/73), runners = 46(43/55), rowers = 53(42/77)%, p<0.05]. Neither the sport modality or the training period influenced the indices of heart rate variability at rest and tilt test. The sport modality and the training period influenced the blood pressure and its variability at rest and tilt test. Rowers showed lower spontaneous baroreflex sensitivity than runners at CP (p=0.03). Both the sport modality and the training period influenced the heart rate response in 24 hours, its variability and blood pressure response in 24 hours. CONCLUSION: Both Sport modality and physical training period influenced the cardiac structural, intrinsic and autonomic adaptations as well the vascular adaptations. However, the sport modality seems to influence more significantly these adaptations

INTRODUÇÃO: O treinamento físico provoca adaptações cardiovasculares, sendo que maiores adaptações estruturais cardíacas são observadas em atletas de elite, que realizam treinamento físico de alta intensidade com o objetivo de aumento no rendimento. Além disto, atletas apresentam diminuição da frequência cardíaca de repouso, embora os mecanismos que explicam a bradicardia em atletas ainda são controversos. O nível das adaptações pode variar e fatores como o tipo de modalidade esportiva, tempo e nível de treinamento físico podem contribuir para tal variação. Desta forma, o objetivo deste estudo foi investigar a influência de modalidades esportivas (ciclismo, corrida de longa distância e remo) e da periodização do treinamento físico nas adaptações estruturais e funcionais, autonômicas e não autonômicas cardíacas e vasculares em atletas de elite no repouso, na inclinação ortostática e nas 24 horas. MÉTODOS: Neste estudo experimental longitudinal prospectivo foram avaliados 13 ciclistas, 13 corredores e 11 remadores de elite, saudáveis (entre 20 e 36 anos; masculino), engajados em treinamento físico competitivo, em 2 períodos de treinamento: período básico-PB e período competitivo-PC. Avaliação da capacidade funcional máxima foi feita por teste cardiorrespiratório. Adaptações estruturais cardíacas foram avaliadas por meio do ecocardiograma bidimensional com doppler. Frequência cardíaca intrínseca foi estudada por meio do duplo bloqueio farmacológico (atropina 0,04 mg/kg e esmolol 500 g/kg, i.v.). Frequência cardíaca e pressão arterial foram registradas continuamente no repouso e no teste de inclinação ortostática por meio de ECG e monitor de pressão arterial, respectivamente (500Hz). A variabilidade da frequência cardíaca e pressão arterial foram analisadas pelo método auto-regressivo. Frequência cardíaca e pressão arterial de 24 horas foram aquisitadas pelo Holter e Mapa, respectivamente. Avaliação da sensibilidade barorreflexa espontânea foi calculada pelo método da sequência. Os dados foram apresentados como mediana e variação interquartil. RESULTADOS: Remadores apresentaram maior VO2max que os corredores no PC (p<0,04). Ciclistas e remadores apresentaram maior VO2max no PC comparado ao PB (p<0,05). Corredores apresentaram maiores diâmetros diastólicos do ventrículo esquerdo que ciclistas (p<0,06) e remadores (p<0,01) no PB. Corredores e ciclistas apresentaram maiores diâmetros diastólicos do ventrículo esquerdo que remadores (p<0,004) no PC. Corredores apresentaram maiores índices de massa do ventrículo esquerdo que ciclistas (p<0,04) no PB e ciclistas maiores que remadores (p<0,03) no PC. Ciclistas foram os únicos atletas que apresentaram a reversão do remodelamento cardíaco no PB (p<0,04). No repouso, corredores mostraram menor frequência cardíaca que ciclistas no PC ciclistas = 50(45/55), corredores = 44(43/47), remadores = 44(43/53)bpm, p<0,03]. Corredores e remadores mostraram maior efeito vagal [ciclistas = 41(36/46), corredores = 55(48/59), remadores = 50(42/66)bpm, p=0,03] e maior frequência cardíaca intrínseca ciclistas = 84(82/87), corredores = 92(87/94), remadores = 96(85/101)bpm, p=0,03] que ciclistas no PC. Remadores tinham maior frequência cardíaca intrínseca que ciclistas no PB ciclistas = 88(86/92), corredores = 91(82/99), remadores = 95(90/101)bpm, p=0,03]. PC comparado ao PB diminuiu o efeito vagal 41(36/46) vs. 48(43/51)bpm, p<0,05] e frequência cardíaca intrínseca 84(82/87) vs. 88(86/92), p<0,05] dos ciclistas. No teste de inclinação, corredores mostraram menor aumento da frequência cardíaca que ciclistas no PB ciclistas = 64(49/73), corredores = 46(43/55), remadores = 53(42/77)%, p<0,05]. Nem a modalidade ou período de treinamento físico influenciaram os índices de variabilidade da frequência cardíaca no repouso ou na inclinação ortostática. A modalidade esportiva e o período de treinamento físico influenciaram a pressão arterial e sua variabilidade, no repouso e na inclinação ortostática. Remadores apresentaram menor sensibilidade barorreflexa espontânea que corredores no PC (p=0,03). Tanto a modalidade quanto o período de treinamento físico influenciaram a resposta da frequência cardíaca de 24 horas, sua variabilidade e a resposta da pressão arterial de 24 horas. CONCLUSÃO: Tanto a modalidade esportiva quanto o período de treinamento físico influenciaram nas adaptações estruturais cardíacas, intrínsecas e autonômicas cardíacas e vasculares. Entretanto a modalidade esportiva parece influenciar mais expressivamente essas adaptações / INTRODUCTION: Physical exercise training provokes cardiovascular adaptations and the highest structural cardiac adaptations are observed in elite athletes who perform high intensity training with the objective of increasing their physical performance. Besides, athlete shows decrease on resting heart rate. However, the mechanisms that explain the bradycardia in athletes are still controversial. The level of these adaptations may vary and some factors as the type of sport modality, time and level of physical training can contribute to such different responses. Thus, the aim of this study was to investigate the influence of sport modalities (cycling, long distane runner and rower) and physical training periodization on cardiac structural and functional, cardiac autonomic and non-autonomic and vascular adaptations in elite athletes at rest, tilt table test and within 24 hours. METHODS: In this prospective longitudinal experimental study, 13 cyclists, 13 runners and 11 rowers, healthy (20 to 36 years old; male), engaged in competitive training were evaluated in 2 periods of training: basic period BP and competitive period -CP. Maximal functional capacity was evaluated by cardiopulmonary test. Cardiac structural adaptations were evaluated by two-dimensional echocardiography with doppler. Intrinsic heart rate was studied by means of double pharmacological blockade (atropine 0.04 mg/kg and esmolol 500 g/kg, iv.). Heart rate and blood pressure were recorded continuously at rest and tilt table test by means of ECG and arterial blood pressure monitor, respectively (500Hz). The heart rate and blood pressure variabilities were analyzed by autoregressive method. Heart rate and blood pressure within 24 hours were recorded using Holter and blood pressure ambulatory monitor, respectivamente. Spontaneous baroreflex sensitivity was calculated using the sequence method. The data were presented as median and interquartile range. RESULTS: Rowers showed higher VO2max than runners at CP (p<0.04). Cyclists and rowers showed higher VO2max at CP compared to BP (p<0.05). Runners presented higher left ventricular diastolic diameters than cyclists (p<0.06) and rowers (p<0.01) at BP. Runners and cyclists presented higher left ventricular diastolic diameters than rowers (p<0.004) at CP. Runners showed higher left ventricular mass index than cyclistas (p<0.04) at BP and cyclists higher than rowers (p<0.03) at CP. Cyclists were the only athletes who had a reversal of cardiac remodeling at BP (p<0.04). At rest, runners showed lower heart rate than cyclists at CP cyclists = 50(45/55), runners = 44(43/47), rowers = 44(43/53)bpm, p<0.03]. Runners and rowers showed higher vagal effect [cyclists = 41(36/46), runners = 55(48/59), rowers = 50(42/66)bpm, p=0.03] and higher intrinsic heart rate cyclists = 84(82/87), runners = 92(87/94), rowers = 96(85/101)bpm, p=0,03] than cyclists at CP. Rowers had higher intrinsic herat rate than cyclists at BP cyclists = 88(86/92), runners = 91(82/99), rowers = 95(90/101)bpm, p=0.03]. CP compared with BP decreased the vagal effect 41(36/46) vs. 48(43/51)bpm, p<0.05] and intrinsic heart rate 84(82/87) vs. 88(86/92), p<0.05] of cyclists. At tilt table test, runners showed smaller increase in heart rate than cyclists at BP cyclists = 64(49/73), runners = 46(43/55), rowers = 53(42/77)%, p<0.05]. Neither the sport modality or the training period influenced the indices of heart rate variability at rest and tilt test. The sport modality and the training period influenced the blood pressure and its variability at rest and tilt test. Rowers showed lower spontaneous baroreflex sensitivity than runners at CP (p=0.03). Both the sport modality and the training period influenced the heart rate response in 24 hours, its variability and blood pressure response in 24 hours. CONCLUSION: Both Sport modality and physical training period influenced the cardiac structural, intrinsic and autonomic adaptations as well the vascular adaptations. However, the sport modality seems to influence more significantly these adaptations

No description available.

Heart rate changes mediate the embryotoxic effect of antiarrhythmic drugs in the chick embryo A significant increase in cardiovascular medication use during pregnancy has occurred in recent years but only limited evidence on its safety profile is available. We hypothesized that drug-induced bradycardia is the leading mechanism of developmental toxicity. We tested metoprolol, carvedilol, or ivabradine for embryotoxicity and their acute effect on chick embryonic model. We used video microscopy and ultrasound biomicroscopy. Significant dose-dependent mortality was achieved in embryos injected with carvedilol and ivabradine. In ED4 embryos, metoprolol, carvedilol and ivabradine reduced the heart rate by 33%, 27%, and 55%, respectively, compared to controls (6%). In ED8 embryos this effect was more pronounced with a heart rate reduction by 71%, 54%, 53%, respectively (controls 36%). Cardiac output decreased in all tested groups but only proved significant in the metoprolol group in ED8 embryos. The number of -adrenergic receptors showed a downward tendency during embryonic development but a negative chronotropic effect of tested drugs was increasingly pronounced with embryonic maturity. This effect was associated with reduced cardiac output in chick embryos, probably leading to premature death....

This thesis describes a novel method to reversibly and safely block nerve conduction using a low frequency alternating current (LFAC) waveform at 1 Hz applied through a bipolar extrafascicular electrode. This work follows up on observations made on excised mammalian peripheral nerves and earthworm nerve cords. An<i> in-situ</i> electrophysiology setup was used to assess the LFAC<br>waveform on propagating action potentials (APs) within the cervical vagus nerve in anaesthetized Sprague-Dawley rats (n = 12). Two sets of bipolar cuff or hook electrodes were applied unilaterally to the cervical vagus nerve, which was crushed rostral to the electrodes to exclude reflex effects<br>on the animal. Pulse stimulation was applied to the rostral electrode, while the LFAC conditioning waveform was applied to the caudal electrode. The efferent volley, if unblocked, elicits acute bradycardia and hypotension. The degree of block of the vagal stimulation induced bradycardia<br>was used as a biomarker. Block was assessed by the ability to reduce the bradycardic drive by monitoring the heart rate (HR) and blood pressure (BP) during LFAC alone, LFAC with vagal stimulation, and vagal stimulation alone. LFAC applied via a hook electrode (n = 7) achieved 86.6 +/- 11% block at current levels 95 +/- 38 uAp (current to peak). When applied via a cuff electrode (n = 5) 85.3 +/- 4.60% block was achieved using current levels of 110 +/- 65 uAp. Furthermore, LFAC was explored on larger vagal afferent fibers in larger human sized nerve bundles projecting to effects mediated by a reflex. The effectiveness of LFAC was assessed in an <i>in-situ</i> electrophysiological setup on the left cervical vagus in anaesthetized domestic swine (n = 5). Two bipolar cuff electrodes were applied unilaterally to the cervical vagus nerve, which was crushed caudal to the electrodes to eliminate cardiac effects. A tripolar extrafascicular cuff electrode was placed most rostral on the nerve for recording of propagating APs induced by<br>electrical stimulation and blocked via the LFAC waveform.<br>Standard pulse stimulation was applied to the left cervical vagus to induce the Hering-Breuer reflex. If unblocked, the activation of the Hering-Breuer reflex would cause breathing to slow down and potentially cease. Block was quantified by the ability to reduce the effect of the Hering-Breuer<br>reflex by monitoring the breathing rate during LFAC alone, LFAC and vagal stimulation, and vagal stimulation alone. LFAC achieved 87.2 +/- 8.8% (n = 5) block at current levels of 0.8 +/- 0.3 mAp. Compound nerve action potentials (CNAP) were monitored directly. They show changes<br>in nerve activity during LFAC, which manifests itself as the slowing and amplitude reduction of components of the CNAPs. Since the waveform is balanced, all forward reactions are reversed, leading to a blocking method that is similar in nature to DC block without the potential issues of<br>toxic byproduct production. These results suggest that LFAC can achieve a high degree of nerve block in both small and large nerve bundles, resulting in the change in behavior of a biomarker, <i>in-vivo </i>in the mammalian nervous system at low amplitudes of electrical stimulation that are within the water window of the electrode.<br>

Heart rate changes mediate the embryotoxic effect of antiarrhythmic drugs in the chick embryo A significant increase in cardiovascular medication use during pregnancy has occurred in recent years but only limited evidence on its safety profile is available. We hypothesized that drug-induced bradycardia is the leading mechanism of developmental toxicity. We tested metoprolol, carvedilol, or ivabradine for embryotoxicity and their acute effect on chick embryonic model. We used video microscopy and ultrasound biomicroscopy. Significant dose-dependent mortality was achieved in embryos injected with carvedilol and ivabradine. In ED4 embryos, metoprolol, carvedilol and ivabradine reduced the heart rate by 33%, 27%, and 55%, respectively, compared to controls (6%). In ED8 embryos this effect was more pronounced with a heart rate reduction by 71%, 54%, 53%, respectively (controls 36%). Cardiac output decreased in all tested groups but only proved significant in the metoprolol group in ED8 embryos. The number of -adrenergic receptors showed a downward tendency during embryonic development but a negative chronotropic effect of tested drugs was increasingly pronounced with embryonic maturity. This effect was associated with reduced cardiac output in chick embryos, probably leading to premature death....