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Application of Periodic Acid-Schiff Fluorescence Emission for Immunohistochemistry of Living Mouse Renal Glomeruli

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In Vivo Cryotechnique in Biomedical Research and Application for Bioimaging of Living Animal Organs

Abstract

Glomerular basement membrane (GBM) plays a critical role in preventing serum protein leakage into Bowman’s space. By using “in vivo cryotechnique (IVCT),” the periodic acid-Schiff (PAS) fluorescence emission was well-represented GBM instead of immunostaining of collagen type IV, which was difficult to observe without the microwave treatment in specimens. Serum protein distribution in living mouse glomeruli was better visualized with IVCT compared with other conventional methods. Under normal condition, immunoreaction products of albumin and immunoglobulin G heavy and light chains (IgG (H+L)) were localized within glomerular capillary loops (GCL) but not colocalized with the PAS fluorescence emission of the GBM. Under heart-arrest condition and with quick-freezing of resected tissues, albumin, IgG (H+L), immunoglobulin kappa light chain, and IgG1 heavy chain (IgG1) were immunolocalized within GCL and mesangial areas, but only albumin and kappa light chain were additionally immunolocalized in Bowman’s space, indicating their passage through GBM. Under acute hypertensive condition, both albumin and kappa light chain, but not IgG1, were clearly immunolocalized along GBM and in Bowman’s space, indicating their increased passage through GBM. The overlapped areas of PAS fluorescence emission and albumin or kappa light chain were appeared to be larger with quick-freezing and under the heart-arrest or acute hypertensive condition than under normal circulation, whereas those of PAS emission and IgG1 were not different among those conditions. These results suggest that PAS fluorescence emission combined with IVCT allowed us to investigate the serum proteins passing through GBM induced by hemodynamic changes.

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Authors and Affiliations

  1. Department of Nephrology, The First Hospital of China Medical University, No.155 North Nanjing Street, Heping District, Shenyang, Liaoning, 110001, China

    Zilong Li M.D., Ph.D.

  2. Department of Anatomy, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo City, Yamanashi, 409-3898, Japan

    Zilong Li M.D., Ph.D. & Shinichi Ohno

  3. Division of Health Sciences, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto City, Nagano, 390-8621, Japan

    Nobuo Terada

  4. Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokato, Chuo City, Yamanashi, 409-3898, Japan

    Nobuo Terada

Authors
  1. Zilong Li M.D., Ph.D.
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  2. Nobuo Terada
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  3. Shinichi Ohno
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Correspondence to Zilong Li M.D., Ph.D. .

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Editors and Affiliations

  1. Department of Anatomy and Molecular Histology Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Yamanashi, Japan

    Shinichi Ohno

  2. Department of Anatomy and Molecular Histology Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Yamanashi, Japan

    Nobuhiko Ohno

  3. Division of Health Science, Shinshu University Graduate School of Medicine, Nagano, Japan

    Nobuo Terada

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Li, Z., Terada, N., Ohno, S. (2016). Application of Periodic Acid-Schiff Fluorescence Emission for Immunohistochemistry of Living Mouse Renal Glomeruli. In: Ohno, S., Ohno, N., Terada, N. (eds) In Vivo Cryotechnique in Biomedical Research and Application for Bioimaging of Living Animal Organs. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55723-4_22

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  • DOI: https://doi.org/10.1007/978-4-431-55723-4_22

  • Publisher Name: Springer, Tokyo

  • Print ISBN: 978-4-431-55722-7

  • Online ISBN: 978-4-431-55723-4

  • eBook Packages: MedicineMedicine (R0)

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