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Journal of Histochemistry and Cytochemistry, Vol. 47, 525-532, April 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Ultrastructural Localization of S100A3, a Cysteine-rich, Calcium Binding Protein, in Human Scalp Hair Shafts Revealed by Rapid-freezing Immunocytochemistry

Takami Takizawaa,b, Toshihiro Takizawaa,b, Seiichi Araib, Kenji Kizawac, Hideyo Uchiwac, Ichiro Sasakic, and Takafumi Inouec
a Department of Anatomy, Jichi Medical School, Tochigi
b Cosmetics Laboratory, Kanebo Ltd., Odawara, Kanagawa, Japan
c Basic Research Laboratory, Kanebo Ltd., Odawara, Kanagawa, Japan

Correspondence to: Toshihiro Takizawa, Dept. of Anatomy, Jichi Medical School, 3311 Yakushiji, Minamikawachi-machi, Tochigi 329-0498, Japan.

We have characterized the subcellular distribution of S100A3, a cysteine-rich calcium binding protein, in human scalp hair shaft. This was accomplished using rapid-freezing immunocytochemistry, a technique that combines rapid-freezing, freeze-substitution fixation without chemical fixatives, and subsequent electron microscopic detection of immunocytochemical labeling. This technique preserves both the antigenicity and the ultrastructural integrity of fully keratinized tissues, which are highly unmanageable when prepared for immunoelectron microscopy. In the hair shaft, S100A3 was primarily identified in the endocuticle and was also present in the intermacrofibrillar matrix surrounding macrofibril bundles of intermediate filament keratins in cortex cells. Double immunolabeling of S100A3 and hair keratins revealed the in situ spatial relationship between them. In the endocuticle, S100A3 was present on the inner portion of the endocuticle adjacent to the cell membrane complex, whereas hair keratins were present on the outer portion. These results provide the first ultrastructural evidence that an S100 protein is localized in specific subcompartments in human hair cells. (J Histochem Cytochem 47:525–532, 1999)

Key Words: S100 proteins, S100A3, cysteine-rich calcium binding, proteins, hair keratins, human scalp hair shafts, cortex, cuticle, rapid-freezing, immunocytochemistry, electron microscopy


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