Detection of glycosaminoglycans in the Golgi complex of chondrocytes.
Elongation and sulfation of glycosaminoglycans are pivotal roles of the Golgi complex during the biosynthesis of proteoglycan monomers. In the present work the spatial relationship between these processes has been investigated by using a combination of immunocytochemical and cytochemical techniques. Chondroitin sulfate and keratan sulfate glycosaminoglycans were immunocytochemically localized in 1 to 2 transmost cisternae, also in a system of narrow tubules at the trans face of the Golgi complex of chick epiphyseal chondrocytes. At these same locations sulfate groups were revealed with the high iron diamine (HID) method, proteoglycan monomers being visualized with ruthenium red. Several treatments were assayed in order to reversibly block the secretory pathway. Chondrocytes incubated at a low temperature, 15 degrees C, before fixation, showed both glycosaminoglycans in the middle cisternae of the Golgi stack as well as the above mentioned locations. After low temperature treatment both HID and ruthenium red stained the middle, but not the cis cisternae. Incubation of the cells for 30 min with either diethylcarbamazine or monensin before fixation permitted detection of glycosaminoglycans and proteoglycan monomers in the middle cisternae, whereas HID staining of the Golgi complex, but not that of secretory vesicles, was abolished. The results show that elongation of both chondroitin sulfate and keratan sulfate glycosaminoglycans takes place in the same Golgi compartments. These include the middle cisternae and probably also the trans cisternae and tubules. Also suggested is that sulfation of one or both types of glycosaminoglycans begins in the middle cisternae.
Velasco
A
A
Department of Cell Biology, Faculty of Biology, University of Seville, Spain.
Hidalgo
J
J
Pérez-Vilar
J
J
García-Herdugo
G
G
Navas
P
P
D000818:Animals; D002645:Chickens; D002807:Chondroitin / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D006132:Growth Plate / Q000166:cytology* / Q000378:metabolism; D007150:Immunohistochemistry; D007632:Keratan Sulfate / Q000378:metabolism; D008854:Microscopy, Electron; D011509:Proteoglycans / Q000378:metabolism; D013431:Sulfates / Q000378:metabolism
D006025:Glycosaminoglycans; D011509:Proteoglycans; D013431:Sulfates; D002807:Chondroitin; D007632:Keratan Sulfate