Incorporation of labeled glucosamine into glycoproteins by organ cultures of hamster trachea: adverse effects of HEPES buffer.
Hamster tracheas were cultured in serum-free CMRL 1066 medium buffered with either NaHCO3 alone or HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulphonic acid) plus NaHCO3 in an atmosphere of 95% O2 and 5% CO2. Afther 2 days in culture, tracheas maintained in HEPES plus NaHCO3-buffered medium showed an altered surface morphology. Histiological examination after 6 days of culture with HEPES plus NaHCO3 showed the presence of only scattered clumps of cilia. Incorporation of [14C]-glucosamine into intracellular glycoproteins was reduced by 75% and into secreted glycoproteins by 54% in cultures buffered with HEPES plus NaHCO3, compared to NaHCO3-buffered cultures. Incorporation of [H]fucose into intracellular glycoproteins was also reduced, although no effect was observed on secreted glycoproteins.
D000818:Animals; D001639:Bicarbonates / Q000494:pharmacology; D002021:Buffers; D002923:Cilia / Q000502:physiology; D006224:Cricetinae; D003470:Culture Media; D004847:Epithelial Cells; D005643:Fucose / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism*; D006020:Glycopeptides / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis / Q000096:biosynthesis*; D006531:HEPES / Q000494:pharmacology*; D009068:Movement; D009924:Organ Culture Techniques; D010879:Piperazines / Q000494:pharmacology*; D014132:Trachea / Q000166:cytology / Q000378:metabolism*
D001639:Bicarbonates; D002021:Buffers; D003470:Culture Media; D006020:Glycopeptides; D006023:Glycoproteins; D010879:Piperazines; D005643:Fucose; D005944:Glucosamine; D006531:HEPES