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  • Title: Major influence of cell differentiation status on characteristics of proteoglycans synthesized by cultured rabbit renal proximal tubule cells: role of insulin and dexamethasone.
    Author: Lelongt B, Vandewalle A, Brenchley PE, Baudouin B, Géniteau-Legendre M, Verroust PJ, Ronco PM.
    Journal: J Cell Physiol; 1993 Jan; 154(1):175-91. PubMed ID: 8419403.
    Abstract:
    To analyze the influence of epithelial cell differentiation and the effects of hormones on the characteristics of cell-associated and secreted proteoglycans (PGs), we studied their distribution, synthesis, and biochemical features in a model of renal proximal tubule cells in primary culture in which cell differentiation could be controlled by medium composition. In cells cultured in serum-free, hormonally defined medium supplemented with insulin and dexamethasone that exhibited a high degree of morphological and functional proximal differentiation (Ronco et al., 1990), cell-associated PGs were similar to those extracted in vivo by their size estimated by Sepharose CL-6B chromatography (Kav = 0.27, vs. 0.26), composition (heparan-sulfate), and localization in a continuous basal layer of extra-cellular matrix (ECM). In contrast, major quantitative and qualitative anomalies of cell-associated PGs were observed in poorly differentiated cells grown in 1% fetal calf serum-supplemented medium (FCS). PGs alterations included: (1) reduced and irregular expression of PGs at the cell basal pole, (2) a 2.8-fold decrease in [35S]-sulfate incorporation into cell-associated PGs, (3) a 3.1-fold increase in trypsin-releasable PGs, and (4) the emergence of a high MW PG composed exclusively of chondroitin-sulfate (CS) (Kav = 0.09 on Sepharose CL-6B) as well as of putative free CS-glycosaminoglycan (GAG) chains (Kav = 0.49 on Sepharose CL-6B). The same alterations were identified in the basal defined medium devoid of hormones but were partially or totally abolished by addition of insulin and dexamethasone, respectively. At variance with cell-associated PGs, production and biochemical features of secreted PGs were not influenced by cell differentiation status and medium composition.
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