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Ferritin immunohistochemistry as a marker for microglia

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Summary

An immunohistochemical analysis of formalin-fixed, paraffin-embedded brain sections was performed with antisera against holoferritin and the light(L)-subunit of ferritin. Sections immunostained using anti-glial fibrillary acidic protein (GFAP), Ricinus communis agglutinin-1 (RCA-1) stain for microglia and iron stain (Berlin blue stain) were compared. The L-subunit of ferritin was purified from normal human spleen according to the modified scrapie-associated fibrils purification, and the antiserum was raised in a rabbit. Both ferritin antisera positively stained resting and, more markedly, reactive microglia, both of which were also stained with RCA-1 but not with GFAP. Ferritin-positive resting microglia were seen more abundantly in cerebral and cerebellar cortices than in white matter. The advantages of ferritin antisera over RCA-1 are as follows. (1) RCA-1 heavily stains blood vessels, while anti-ferritin does not, hence the microglial cells are more readily visualized with ferritin immunohistochemistry. (2) Reactive microglia and macrophages are more strongly stained with anti-ferritin. (3) The staining intensity of ferritin is independent of the length of tissue fixation in formalin. However, anti-ferritin is inferior to RCA-1 in staining resting microglia with a scanty cytoplasm, especially in the white matter, probably because the former recognizes cytoplasmic components, while the latter recognizes cell membrane. Iron stain only gave a reaction to microglial cells in brains with neurosyphilis and to hemosiderin-laden macrophages. Thus, in addition to RCA-1, ferritin antisera are useful as a microglia marker in formalin-fixed, paraffin-embedded sections.

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Authors and Affiliations

  1. Department of Neuropathology, Neurological Institute, Faculty of Medicine, Kyushu University 60, 812, Fukuoka, Japan

    Y. Kaneko, T. Kitamoto & J. Tateishi

  2. Department of Neurosurgery, Neurological Institute, Faculty of Medicine, Kyushu University 60, 812, Fukuoka, Japan

    Y. Kaneko

  3. Tokyo Research Laboratories, Kyowa-Hakko Kogyo co, 3-6-6 Asahi machi, 194, Machida, Tokyo, Japan

    K. Yamaguchi

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  1. Y. Kaneko
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  2. T. Kitamoto
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  3. J. Tateishi
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  4. K. Yamaguchi
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Supported in part by Dr. A. Kondo, Department of Neuropathology, Neurological Institute, Kyushu University

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Kaneko, Y., Kitamoto, T., Tateishi, J. et al. Ferritin immunohistochemistry as a marker for microglia. Acta Neuropathol 79, 129–136 (1989). https://doi.org/10.1007/BF00294369

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  • DOI: https://doi.org/10.1007/BF00294369

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