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Journal of Histochemistry and Cytochemistry, Vol. 47, 881-888, July 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Sulfated Glycosaminoglycans In Guinea Pig Neutrophils Studied by Use of Cationic Colloidal Gold

Dong-Hua Yanga, Shinichiro Tsuyamaa, Jun Ohmoria, and Fusayoshi Murataa
a Department of Anatomy, Faculty of Medicine, Kagoshima University, Kagoshima, Japan

Correspondence to: Fusayoshi Murata, Dept. of Anatomy, Faculty of Medicine, Kagoshima Univ., 8-35-1 Sakuragaoka, Kagoshima 890-8520, Japan.

Using a high electron resolution staining method, cationic colloidal gold (CCG, pH 1.0) staining, we studied the fine structural localization of sulfated glycosaminoglycans (GAGs) in various maturational stages of guinea pig neutrophils. Azurophil and specific granules of neutrophils reacted positively to CCG, with variety in labeling according to maturation. All immature azurophil and specific granules were labeled selectively. Mature granules lost their affinity with CCG. CCG-positive labeling was also observed in the trans to trans-most Golgi apparatus of promyelocytes and myelocytes. Prior absorption with poly-L-lysine prevented CCG labeling of tissue sections. Mild methylation of ultrathin sections at 37C did not alter CCG labeling, whereas CCG labeling disappeared after active methylation at 60C. Treatment with chondroitinase ABC or heparinase I abolished the majority of CCG labeling. These findings suggest the existence of sulfated GAGs not only in immature azurophil but also in immature specific granules of neutrophils. Sulfation of GAGs occurs in the trans- to trans-most Golgi apparatus of neutrophil granulocytes. A possible correlation between accumulation of sulfated GAGs and maturation of specific granules in neutrophils is also discussed. (J Histochem Cytochem 47:881–887, 1999)

Key Words: neutrophil, sulfated glycosaminoglycan, cationic colloidal gold, azurophil granule, specific granule


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