Demonstration of anionic sites in human eccrine and apocrine sweat glands in post-embedded ultra-thin sections with cationic colloidal gold: effect of enzyme digestion on these anionic sites.
We localized anionic sites ultrastructurally in human eccrine and apocrine sweat glands with a poly-L-lysine-gold complex (cationic colloidal gold). Anionic sites were labeled by incubating Lowicryl K4M-embedded sections on droplets of cationic colloidal gold. In eccrine sweat glands, colloidal gold particles were restricted to the basolateral membrane of the secretory cells at low pH, whereas the luminal membrane did not react with the gold particles. Chondroitinase ABC digested these anionic sites. This indicates that chondroitin sulfate and/or dermatan sulfate constitutes anionic sites in the basal labyrinth of eccrine sweat glands. In apocrine sweat glands, the luminal membrane of the secretory cells showed strong reaction at low pH, whereas the contraluminal membrane did not show any reaction. Neuraminidase completely digested these anionic sites, which indicated that the anionic charge of the apocrine lumen was due to sialic acid. Differences in distribution and susceptibility to enzymes of anionic sites in cell membranes between eccrine and apocrine sweat glands may reflect functional differences between these glands. Dark cell granules in eccrine secretory cells were negative for the anionic sites when sections were labeled without any pre-treatment. However, pre-incubation of the grids on EGTA or deionized water unmasked the anionic sites on the dark cell granules. The positive staining after EGTA treatment was greatly decreased by reincubation with CaCl2. These results suggested that Ca blocked anionic sites in dark cell granules. Exposed anionic sites were digested with chondroitinase ABC. This indicated that chondroitinase ABC and/or dermatan sulfate composed the anionic sites in dark cell granules.
Saga
K
K
Department of Dermatology, Sapporo Medical College, Japan.
Takahashi
M
M
D000838:Anions / Q000032:analysis*; D001665:Binding Sites / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002808:Chondroitin Lyases; D003593:Cytoplasm / Q000378:metabolism; D004533:Egtazic Acid; D006046:Gold; D006801:Humans; D006821:Hyaluronoglucosaminidase; D006863:Hydrogen-Ion Concentration; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase; D011107:Polylysine; D012794:Sialic Acids / Q000032:analysis; D013545:Sweat Glands / Q000378:metabolism* / Q000648:ultrastructure
D000838:Anions; D012794:Sialic Acids; D011107:Polylysine; D004533:Egtazic Acid; D006046:Gold; D009439:Neuraminidase; D006821:Hyaluronoglucosaminidase; D002808:Chondroitin Lyases; D019158:N-Acetylneuraminic Acid; D002118:Calcium