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Journal of Histochemistry and Cytochemistry, Vol. 46, 1091-1096, September 1998, Copyright © 1998, The Histochemical Society, Inc.

TECHNICAL NOTE

5'-Nucleotidase in Rat Photoreceptor Cells and Pigment Epithelial Cells Processed by Rapid-freezing Enzyme Cytochemistry

Toshihiro Takizawaa
a Department of Anatomy, Jichi Medical School, Tochigi, Japan

Correspondence to: Toshihiro Takizawa, Dept. of Anatomy, Jichi Medical School, 3311 Yakushiji, Minamikawachi-machi, Tochigi 329-0498, Japan. E-mail: ttakizawa@jichi.ac.jp.

This report describes the subcellular distribution of 5'-nucleotidase (5'-NT) in rat photoreceptor cells and pigment epithelial cells processed by rapid-freeze enzyme cytochemistry. There was a striking difference in the ultrastructural localization of 5'-NT activity between rod outer segments after freeze-substitution fixation and conventional fixation. By rapid-freezing enzyme cytochemistry, 5'-NT activity was localized in the extradiscal space of intact nonvacuolated discs, whereas by conventional cytochemistry it was shown in the intradiscal space of artifactual vacuolated discs. In the freeze-substituted retinal cells, an appreciable difference in functional 5'-NT molecules was also found. The soluble 5'-NT on the cytoplasmic side of the disc membrane was vital in the rod outer segments, whereas the membrane-bound ecto-5'-NT on the exoplasmic (external) surface of the apical process was active in the pigment epithelial cells. Rapid-freezing enzyme cytochemistry should be worth employing as a method to reveal the fine localization of enzyme activity at the level of cell ultrastructures, which are poorly preserved by conventional fixation, and should provide information approximate to that in living cells. (J Histochem Cytochem 46:1091–1095, 1998)

Key Words: rapid freezing, freeze-substitution fixation, enzyme cytochemistry, 5'-nucleotidase, cGMP-related enzymes, rat retinas, photoreceptor cells, pigment epithelial cells, electron microscopy


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