Binding of lectin-fluorescein conjugates to intracellular compartments of growth-plate chondrocytes in situ.
In this study, lectin-binding techniques are applied to growth-plate cartilage to analyze the intracellular localization of lectin-binding glycoconjugates of chondrocytes in situ. The binding of ten fluorescein-conjugated lectins is analyzed on 1-micron-thick Epon-embedded, nondecalcified sections of growth plates from Yucatan swine. Comparisons are made to intracellular binding in chondrocytes of tracheal, articular, and auricular cartilage. Ear epithelium, tracheal epithelium, and kidney are used as positive control tissues for the specificity of lectin binding. Only the mannose-binding lectins had affinity for the RER and nuclear envelope. Eight lectins reacted within the Golgi complex with characteristic patterns which ranged from localized fine linear strands to extensive vesicular accumulations. When cartilage slabs were exposed before embedment to the ionophore monensin to disrupt intracellular transport through the Golgi, it was possible to define differential subcompartments of the Golgi complex, based upon sites of sugar addition. Also, it was possible to characterize the cytoplasmic deposits of reserve-zone chondrocytes which were positive with concanavalin A as glycogen, based upon their sensitivity to amylase. This method allows resolution at the light-microscopic level of lectin-binding glycoconjugates with localization to specific organelles. Patterns of intracellular binding were consistent with biochemical data relating to the subcellular localization of processing steps of complex carbohydrates prior to secretion.
D000818:Animals; D002356:Cartilage / Q000166:cytology* / Q000378:metabolism; D003208:Concanavalin A / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004838:Epiphyses / Q000166:cytology*; D019793:Fluorescein; D005452:Fluoresceins / Q000378:metabolism*; D006003:Glycogen / Q000378:metabolism; D006056:Golgi Apparatus; D037102:Lectins / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism; D008854:Microscopy, Electron; D008985:Monensin / Q000494:pharmacology; D013552:Swine; D014909:Wheat Germ Agglutinins
D005452:Fluoresceins; D037102:Lectins; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D006003:Glycogen; D008985:Monensin; D008358:Mannose; D019793:Fluorescein