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Comparison of the histology and stiffness of ventricles in Anura of different habitats


Vertebrate hearts have undergone marked morphological and structural changes to adapt to different environments and lifestyles as part of the evolutionary process. Amphibians were the first vertebrates to migrate to land. Transition from aquatic to terrestrial environments required the ability to circulate blood against the force of gravity. In this study, we investigated the passive mechanical properties and histology of the ventricles of three species of Anura (frogs and toads) from different habitats, Xenopus laevis (aquatic), Pelophylax nigromaculatus (semiaquatic), and Bufo japonicus formosus (terrestrial). Pressure-loading tests demonstrated stiffer ventricles of P. nigromaculatus and B. j. formosus compared X. laevis ventricles. Histological analysis revealed a remarkable difference in the structure of cardiac tissue: thickening of the compact myocardium layer of P. nigromaculatus and B. j. formosus and enrichment of the collagen fibers of B. j. formosus. The amount of collagen fibers differed among the species, as quantitatively confirmed by second-harmonic generation light microscopy. No significant difference was observed in cardiomyocytes isolated from each animal, and the sarcomere length was almost the same. The results indicate that the ventricles of Anura stiffen during adaptation to life on land.

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Data availability

Data are included in the electronic supplementary material.


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The authors acknowledge Amphibian Research Center of Hiroshima University for providing B. j. formosus.


This study was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI (Grant Number JP17H04740 and JP19K22962 to Y. U., JP18K12055 to M. N.) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.

Author information




M. I., Y.U., and M.N. planned and designed the study and wrote the manuscript. M. I. performed the majority of experiments. S.S. provided the technique to observe SHG light from collagen. All authors have read and reviewed the manuscript, and gave final approval for publication.

Corresponding author

Correspondence to Yoshihiro Ujihara.

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All animal experiments were conducted under the guidelines specified by the Guide for Animal Experimentation, Nagoya Institute of Technology.

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The authors declare no competing interests.

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Ito, M., Ujihara, Y., Sugita, S. et al. Comparison of the histology and stiffness of ventricles in Anura of different habitats. J Biol Phys 47, 287–300 (2021).

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  • Passive mechanical properties
  • Vertebrate heart
  • Amphibian heart
  • Evolution
  • Adaptation
  • Terrestrialization