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Development of a new histological identification method of human sinoatrial node suitable for immunohistochemical study

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Abstract

Histological identification of the human sinoatrial node (SAN) remains a challenge. Conventional identification methods, such as Lev’s method, have certain limitations. The aim of our study was to develop a new histological identification method that could properly identify the sinoatrial node, applicable to the immunohistochemical study of intra-nodal structures. Thirty-nine human autopsied hearts were included in this study. The cases included 23 men and 16 women ranging in age from 20 to 99 years. The sinoatrial area from eight control samples was cut in the vertical section using the conventional Lev’s method. In our new method, called the “En face one-block method,” the sinoatrial node was cut in “En face” at the junction of the right border of the right appendage and superior vena cava, placed in one long cassette, and serially cut using a microtome. Immunostaining was performed using primary antibodies against CD31, podoplanin (D2-40), S-100, and other proteins. The average area of the SAN on the slide glass in our new method was 32.2 mm2, which was significantly larger than that (3.59 mm2) of the control samples by Lev’s method. The SAN area was positively correlated with age (r = 0.357; p = 0.026), especially in women (r = 0.626; p = 0.0095). The SAN group had significantly lower percentage of CD31-positive blood capillaries, higher percentage of podoplanin-positive lymphatic channels, and S-100-positive peripheral nerves. We successfully developed a novel cutting method applicable to immunohistochemical studies, with which we could provide a bird’s-eye view of the sinoatrial nodes.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

I would like to thank Mr. Yasuhiro Enoki, MSc, and Ms. Sujata Shaka, Ph.D., and the pathological staff who performed autopsies at TMDU Medical Hospital. I appreciate Dr. Satoru Muro and Professor Keiichi Akita, the Department of Clinical Anatomy, TMDU, for providing immense help on the 3D reconstruction of the SAN. The authors sincerely thank those who donated their bodies to science so that anatomical research could be performed. The results from such research increase our knowledge, which can then improve patient care. Therefore, these donors and their families deserve our highest gratitude.

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No financial support was given to this study.

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Author notes

  1. Motoji Sawabe

    Present address: Department of Diagnostic Pathology, Hitachi General Hospital, 2-1-1 Jonancho, Hitachi, Ibaraki, 317-0077, Japan

Authors and Affiliations

  1. Department of Molecular Pathology, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo, Tokyo, 113-8510, Japan

    Thavisouk Hatthakone, Sunti Oundavong, Yurie Soejima & Motoji Sawabe

  2. Department of Molecular Pathology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo, Tokyo, 113-8519, Japan

    Motoji Sawabe

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  1. Thavisouk Hatthakone
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Hatthakone, T., Oundavong, S., Soejima, Y. et al. Development of a new histological identification method of human sinoatrial node suitable for immunohistochemical study. Anat Sci Int 98, 293–305 (2023). https://doi.org/10.1007/s12565-022-00697-0

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