Histochemistry and Cell Biology

, Volume 133, Issue 2, pp 229–239

Histochemical approach of cryobiopsy for glycogen distribution in living mouse livers under fasting and local circulation loss conditions

  • Yurika Saitoh
  • Nobuo Terada
  • Sei Saitoh
  • Nobuhiko Ohno
  • Yasuhisa Fujii
  • Shinichi Ohno
Original Paper

DOI: 10.1007/s00418-009-0663-4

Cite this article as:
Saitoh, Y., Terada, N., Saitoh, S. et al. Histochem Cell Biol (2010) 133: 229. doi:10.1007/s00418-009-0663-4

Abstract

Soluble proteins and glycogen particles, which are easily lost upon conventional chemical fixation, have been reported to be better preserved in paraffin-embedded sections by ‘cryobiopsy’ combined with freeze-substitution fixation (FS). In this study, we examined the distribution of glycogen in living mouse livers under physiologic and pathologic conditions with periodic acid-Schiff (PAS) staining by cryobiopsy. The livers of the fully fed mice showed high PAS-staining intensity in the cytoplasm of all hepatocytes. The PAS-staining intensity gradually decreased away from hepatocytes around portal tracts, depending on treatments with different α-amylase concentrations. At 6 or 12 h after fasting, PAS-staining intensity markedly decreased in restricted areas of zone I near the portal tracts. The cryobiopsy was repeatedly performed not only on different mice, but also on individuals. Next, glycogen distributions were evaluated by temporarily clipping of liver tissues of anesthetized mice, followed by recovery of blood circulation. In the liver tissues in which blood was recirculated for 1 h after the 30 min anoxia, PAS staining was still observed in zone II and also in restricted areas of zone I far from the portal tracts. In PAS-unstained hepatocytes, the immunoglobulin-kappa light chain was not detected in the cytoplasm, indicating that cell membrane permeability was retained and that glycogen metabolism was related to the functional state of blood circulation. We propose that the level of consumption or production of glycogen particles could vary in zone I, depending on the distance from the portal tracts. Thus, cryobiopsy combined with FS enabled us to examine time-dependent changes in glycogen distribution in the liver tissues of living mice. This combination might be applicable to the clinical evaluation of human liver tissues.

Keywords

CryobiopsyGlycogenPAS stainingZonal difference

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Yurika Saitoh
    • 1
  • Nobuo Terada
    • 1
  • Sei Saitoh
    • 1
  • Nobuhiko Ohno
    • 1
  • Yasuhisa Fujii
    • 1
  • Shinichi Ohno
    • 1
  1. 1.Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and EngineeringUniversity of YamanashiChuo-cityJapan