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Induction of N‐Glycosylation Activity in Cultured Embryonic Rat Brain Cells
Author(s) -
Bhat Narayan R.,
Waechter C. J.
Publication year - 1988
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1988.tb02922.x
Subject(s) - glycosylation , dolichol , glycoprotein , mannose , biology , oligosaccharide , biochemistry , cellular differentiation , cell culture , n linked glycosylation , neurite , microbiology and biotechnology , in vitro , enzyme , biosynthesis , gene , genetics , glycan
Developmental changes in protein N‐glycosylation activity have been studied using cultures of dissociated fetal rat brain cells as an in vitro model system. These cultures undergo an initial phase of neurite outgrowth and cell proliferation (4–6 days in culture), followed by a period of cellular differentiation. N‐Glycosylation activity has been measured by assaying the incorporation of [2‐ 3 H]‐mannose into dolichol‐linked oligosaccharides and glyco‐protein over a period of 1–25 days in culture. This study revealed a marked induction of N‐glycosylation activity beginning at approximately 1 week of culture. [2‐ 3 H]‐Mannose incorporation into the oligosaccharide‐lipid intermediate fraction and glycoprotein reached maximal values between 12 and 16 days of culture and declined thereafter. The major dolichol‐linked oligosaccharide labeled by the brain cell cultures was shown to be Glc 3 Man9GlcNAc 2 by HPLC analysis. Parallel incorporation studies with [ 3 H]leucine showed that the increase in protein N‐glycosylation was relatively higher than a concurrent increase in cellular protein synthesis observed during the induction period. Maximal labeling of glycoprotein corresponded to the period of glial differentiation, as indicated by a sharp rise in the marker enzymes, 2′,3′‐cyclic nucleotide 3′‐phosphohydrolase (an oligodendroglial marker) and glutamine synthetase (an astroglial marker). The results describe a developmental activation of the N‐glycosylation pathway and suggest a possible relationship between N‐linked glycoprotein assembly and the growth and differentiation of glial cells.