Cardiac Enlargement in the Chick Embryo Induced by Hypothermic Incubation Is Due to a Combination of Hyperplasia and Hypertrophy of Cardiomyocytes.

Hypothermic incubation of chicken eggs leads to smaller embryos with enlarged hearts, originally described as hypertrophic. Over the years, however, accumulated evidence suggested that hyperplasia, rather than hypertrophy, is the predominant mechanism of cardiac growth during the prenatal period. We have thus set to re-evaluate the hypothermia model to precise the exact cellular mechanism behind cardiac enlargement. Fertilized chicken eggs were incubated at either 37.5 °C (normothermia) or 33.5 °C from embryonic day (ED) 13 onward (hypothermia). Sampling was performed at ED17, at which point wet embryo and heart weight were recorded, and the hearts were submitted to histological examination. In agreement with previous results, the hypothermic embryos were 29% smaller and had hearts 18% larger, translating into a 67% increase in the heart to body weight ratio (P < 0.05 for all parameters). The cell size was essentially the same between control and hypothermic hearts in all regions analysed. Likewise, there was no significant relationship between the cell size and heart weight; however, in the hypothermic hearts, there was a trend showing positive correlation between cell sizes in different cardiac regions and heart weight. Proliferation rate, determined on the basis of anti-phosphohistone H3 immunofluorescence, showed an overall increase in the hypothermic group, reaching statistical significance (P = 0.02, t-test) in the right ventricle. The proliferation rate was similar among different regions of the same heart. However, the correlation between the proliferation rate and heart weight was only small (r2 = 0.007 and r2 = 0.234 for the normothermic and hypothermic group, respectively). We thus conclude that hyperplasia is the predominant response mechanism in this volume-overload model; mechanistically, decreased heart rate at lower temperature increases the end-diastolic and stroke volume, minimizing the drop in cardiac output through the Frank- Starling mechanism.

Acute temperature effects on function of the chick embryonic heart.

AIM: We analysed the effects of acute temperature change on the beating rate, conduction properties and calcium transients in the chick embryonic heart in vitro and in ovo. METHODS: The effects of temperature change (34, 37 and 40 °C) on calcium dynamics in isolated ED4 chick hearts in vitro were investigated by high-speed calcium optical imaging. For comparison and validation of in vitro measurements, experiments were also performed in ovo using videomicroscopy. Artificial stimulation experiments were performed in vitro and in ovo to uncover conduction limits of heart segments. RESULTS: Decrease in temperature from 37 to 34 °C in vitro led to a 22% drop in heart rate and unchanged amplitude of Ca(2+) transients, compared to a 25% heart rate decrease in ovo. Increase in temperature from 37 to 40 °C in vitro and in ovo led to 20 and 23% increases in heart rate, respectively, and a significant decrease in amplitude of Ca(2+) transients (atrium -35%, ventricle -38%). We observed a wide spectrum of arrhythmias in vitro, of which the most common was atrioventricular (AV) block (57%). There was variability of AV block locations. Pacing experiments in vitro and in ovo suggested that the AV blocks were likely caused by relative tissue hypoxia and not by the tachycardia itself. CONCLUSION: The pacemaker and AV canal are the most temperature-sensitive segments of the embryonic heart. We suggest that the critical point for conduction is the connection of the ventricular trabecular network to the AV canal.