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Immunocytochemical localization of 3‐methylcrotonyl‐CoA carboxylase in cultured ependymal, microglial and oligodendroglial cells
Author(s) -
Murín Radovan,
Verleysdonk Stephan,
Rapp Mirna,
Hamprecht Bernd
Publication year - 2006
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.1471-4159.2006.03819.x
Subject(s) - immunocytochemistry , ependymal cell , biology , blot , neuroglia , cell , microbiology and biotechnology , biochemistry , immunohistochemistry , central nervous system , gene , immunology , neuroscience , endocrinology
To evaluate the ability of ependymal, microglial and oligodendroglial cells to degrade leucine, the presence of 3‐methylcrotonyl‐CoA carboxylase (MCC) was investigated in cultures of these cells. MCC is a biotin‐containing heterodimeric enzyme that is specific for the irreversible part of the leucine catabolic pathway. It has been reported previously that in cell culture MCC is expressed in astrocytes and a subpopulation of neurones. In the present study ependymal, microglial and oligodendroglial cell cultures, derived from the brains of newborn rats, were examined for the expression of MCC by RT–PCR, western blotting and immunocytochemistry. The results of RT–PCR and western blotting showed the presence of mRNA as well as protein of both subunits of MCC in ependymal, microglial and oligodendroglial cell cultures. Immunocytochemical investigation of the cellular and subcellular distribution of MCC demonstrated a mitochondrial location of MCC in all neuroglial cell types investigated. The ubiquitous expression of MCC in glial cells demonstrates the ability of the cells to engage in the catabolism of leucine transported into the brain, mainly for the generation of energy.