title: Regular heartbeat rhythm at the heartbeat initiation stage is essential for normal cardiogenesis at low temperature.
abstract: BACKGROUND
  The development of blood flow in the heart is crucial for heart function and embryonic survival. Recent studies have revealed the importance of the extracellular matrix and the mechanical stress applied to the valve cushion that controls blood flow to the formation of the cardiac valve during embryogenesis. However, the events that trigger such valve formation and mechanical stress, and their temperature dependence have not been explained completely. Medaka (Oryzias latipes) inhabits a wide range of East Asia and adapts to a wide range of climates. We used medaka embryos from different genomic backgrounds and analyzed heartbeat characteristics including back-and-forth blood flow and bradyarrhythmia in embryos incubated at low temperature. We also used high-speed imaging analysis to examine the heartbeat of these animals after transient exposure to low temperature.
  
  
  RESULTS
  Embryos of the Hd-rR medaka strain exhibited back-and-forth blood flow in the heart (blood regurgitation) after incubation at 15 °C. This regurgitation was induced by exposure to low temperature around the heartbeat initiation period and was related to abnormalities in the maintenance or pattern of contraction of the atrium or the atrioventricular canal. The Odate strain from the northern Japanese group exhibited normal blood flow after incubation at 15 °C. High-speed time-lapse analysis of the heartbeat revealed that bradyarrhythmia occurred only in Hd-rR embryos incubated at 15 °C. The coefficient of contraction, defined as the quotient of the length of the atrium at systole divided by its length at diastole, was not affected in either strain. The average heart rate after removing the effect of arrhythmia did not differ significantly between the two strains, suggesting that the mechanical stress of individual myocardial contractions and the total mechanical stress could be equivalent, regardless of the presence of arrhythmia or the heart rate. Test-cross experiments suggested that this circulation phenotype was caused by a single major genomic locus.
  
  
  CONCLUSIONS
  These results suggest that cardiogenesis at low temperature requires a constant heartbeat. Abnormal contraction rhythms at the stage of heartbeat initiation may cause regurgitation at later stages. From the evolutionary viewpoint, strains that exhibit normal cardiogenesis during development at low temperature inhabit northern environments.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Watanabe-Asaka
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Sekiya
authors.forename: Hironori
authors.initials: H
authors.lastname: Wada
authors.forename: Takako
authors.initials: T
authors.lastname: Yasuda
authors.forename: Ikuya
authors.initials: I
authors.lastname: Okubo
authors.forename: Shoji
authors.initials: S
authors.lastname: Oda
authors.affiliation: Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, 5-1-5, Kashiwa-no-ha, Kashiwa, Chiba 277-8562, Japan. mitani@k.u-tokyo.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mitani
mesh_terms: D000818:Animals; D001022:Aortic Valve Insufficiency / Q000503:physiopathology; D003080:Cold Temperature; D003326:Coronary Circulation; D005260:Female; D006321:Heart / Q000196:embryology* / Q000503:physiopathology*; D006339:Heart Rate / Q000502:physiology*; D008297:Male; D009200:Myocardial Contraction; D038081:Organogenesis; D009990:Oryzias / Q000145:classification / Q000196:embryology / Q000502:physiology; D012039:Regional Blood Flow; D013045:Species Specificity; D013997:Time Factors