Browsing by Subject "Serum proteins"

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    Immunohistochemical analyses on albumin and immunoglobulin in acute hypertensive mouse kidneys by “in vivo cryotechnique”
    (Murcia : F. Hernández, 2005) Li, Z.; Terada, N.; Ohno, N.; Ohno, S.
    The purpose of this study is to visualize topographical changes of serum proteins, albumin and immunoglobulin, passing through mouse glomerular capillary loops and their reabsorption in renal proximal tubules by immunohistochemistry in combination with our “in vivo cryotechnique”. The “in vivo cryotechnique” was performed on left mouse kidneys under normotensive, experimentally acute hypertensive and heart-arrest conditions. The cryofixed tissues by the technique were routinely processed for freezesubstitution. Serial deparaffinized sections were stained with hematoxylin-eosine and immunostained with antimouse albumin, immunoglobulin G (IgG), kappa or lambda light chain and IgG1 heavy chain antibodies. Under the normotensive and heart-arrest conditions, albumin and IgG were clearly immunolocalized in blood vessels and slightly in apical cytoplasmic parts of some proximal tubules. Under the acute hypertensive condition, the albumin and kappa or lambda light chains, but not IgG1 heavy chain, were strongly immunolocalized in the apical cytoplasm of almost all proximal tubules. This study is the first in vivo visualization for glomerular passage of serum proteins and their transtubular absorption. Thus, the “in vivo cryotechnique” with freeze-substitution can be used for clarifying not only the functional morphology of living animal cells, but also in situ immunohistochemical localization of their components.
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    Immunolocalization of serum proteins in xenografted mouse model of human tumor cells by various cryotechniques
    (Murcia : F. Hernández, 2009) Bai, Y.; Ohno, N.; Terada, N.; Saitoh, S.; Nakazawa, T.; Nakamura, N.; Katoh, R.; Ohno, S.
    The transport mechanism of soluble molecules throughout the interstitial matrix is closely associated with human tumor behavior in vivo. However, the examination of soluble components in histological architectures has been hampered by artifacts caused during conventional tissue preparation. In this study, the immunodistribution of intrinsic and extrinsic serum components in tumor tissues was examined in xenografted human tumor cells using ‘in vivo cryotechnique’ (IVCT) and cryobiopsy, where target tissues are directly cryofixed in vivo. Human lung cancer cells were subcutaneously injected into the dorsal flank of nude mice, and paraffin sections and cryosections of produced tumors were prepared with different methods. Immunolocalization of serum proteins, including albumin, immunoglobulin G (IgG) and IgM, as well as intravenously injected bovine serum albumin (BSA) was examined. Their immunodistribution was more clearly observed in the interstitium by both IVCT and cryobiopsy than conventional methods. IgM was immunolocalized within blood vessels, whereas albumin and IgG were observed in the tumor interstitium. Moreover, intravenously injected bovine serum albumin exhibited leakage from the blood capillaries into surrounding connective tissues in 24 h, but it gradually diffused to the interstitium of the tumor masses during 3 days. These results suggest that molecular leakage from blood capillaries varies significantly in different areas of developing tumors, and that small serum proteins, but not large ones, were abundantly immunolocalized in the tumor interstitium. Both IVCT and cryobiopsy were found to be useful for immunohistochemical studies of soluble molecules in tumors with blood circulation, and may therefore be helpful for further histopathological analyses.