Premium
Constitutive expression of β‐N‐acetylhexosaminidase in a microglial cell line: Transcriptional modulation by lipopolysaccharide and serum factors
Author(s) -
Beccari Tommaso,
Orlacchio Antonio,
Costanzi Egidia,
Appolloni Maria Grazia,
Laurenzi Assunta,
Bocchini Virginia
Publication year - 1997
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/(sici)1097-4547(19971001)50:1<44::aid-jnr5>3.0.co;2-l
Subject(s) - lipopolysaccharide , modulation (music) , cell culture , microbiology and biotechnology , chemistry , line (geometry) , cell , biology , immunology , biochemistry , physics , genetics , mathematics , geometry , acoustics
We investigated the expression of the α‐ and β‐subunits of the lysosomal enzyme β‐N‐acetylhexosaminidase in the BV‐2 microglial cell line under different culture conditions. β‐N‐acetylhexosaminidase from BV‐2 microglia cells was separated into its constituent isoenzymes on diethylaminoethyl (DEAE) cellulose, and its activity was monitored with 4‐methylumbelliferyl‐β‐N‐acetylglucosamine and 4‐methylumbelliferyl‐β‐N‐acetylglucosamine‐6‐sulphate substrates. Forms corresponding to the mouse isoenzymes A and B were present in the cells incubated in serum‐supplemented medium as well as in serum‐free medium. Lipopolysaccharide, a well‐known activator of microglia in vitro, added to the BV‐2 cells in serum‐supplemented medium induced a decrease in the specific enzymatic activity determined with the 4‐methylumbelliferyl‐β‐N‐acetylglucosamine substrate. Lipopolysaccharide had no effect on hexosaminidase isoenzyme pattern of BV‐2 cells in serum‐supplemented medium. The level of α‐subunit mRNA was increased and the level of β‐subunit mRNA was decreased in BV‐2 cells incubated in serum‐supplemented medium plus lipopolysaccharide. In the cells incubated in a serum‐free medium no significant changes in the hexosaminidase‐specific activities towards the above substrates were observed. Interestingly, increased expression of α‐ and β‐subunit mRNA was evident in comparison with cultures in serum‐supplemented medium. The present results suggest that the BV‐2 cell line may be a useful tool to study the possible role of microglia in the metabolism of brain glycolipids. J. Neurosci. Res. 50:44–49, 1997. © 1997 Wiley‐Liss, Inc.