Expression of pyruvate carboxylase in cultured oligodendroglial, microglial and ependymal cells.
The mitochondrial enzyme, pyruvate carboxylase (PC; EC 6.4.1.1) is considered to play a significant role in the intermediary metabolism of neural tissue. PC-catalyzed carboxylation of pyruvate to oxaloacetate is a major anaplerotic reaction in brain. Anaplerosis is essential for homeostasis of the members of the tricarboxylic acid (TCA) cycle. Several biochemical pathways rely on withdrawing TCA cycle members. Prominent among these are biosynthesis of fatty acids and of non-essential amino acids such as aspartate, asparagine, glutamate and glutamine, gluconeogenesis, glycogen synthesis, and regeneration of NADPH. The expression of PC in brain has already been described and assigned to astrocytes. Since pyruvate carboxylase deficiency is associated with malformations of the brain, e.g., inadequate development of the corpus callosum and the lack of myelination, one can hypothesize that PC may be expressed also in glial cells other than astrocytes. Therefore, the expression of PC was investigated in cultured oligodendroglial, microglial, and ependymal cells. As assessed by RT-PCR, all these cultures contain PC mRNA. This mRNA is generated in a transcription process that is regulated by the "distal class" of promoters of the PC gene. The expression of PC among cultured glial cells was studied with a rabbit antiserum by immunoblotting and immunocytochemistry. The results indicate that PC is not only expressed in cultured astroglial cells but also in cultured oligodendrocytes, microglial cells, and ependymocytes. It appears that the intermediary metabolism of these cells includes the anaplerotic action of PC as well as possibly also functions of the enzyme in biosynthetic pathways and the provision of NADPH for defense against reactive oxygen species.
Murin
Radovan
R
Interfaculty Institute for Biochemistry, University of Tuebingen, Hoppe-Seyler-Street 4, 72076 Tuebingen, Germany. radovan.murin@uni-tuebingen.de
Cesar
Marija
M
Kowtharapu
Bhavani S
BS
Verleysdonk
Stephan
S
Hamprecht
Bernd
B
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004805:Ependyma / Q000166:cytology / Q000201:enzymology*; D007150:Immunohistochemistry; D017628:Microglia / Q000201:enzymology*; D009836:Oligodendroglia / Q000201:enzymology*; D011766:Pyruvate Carboxylase / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D011766:Pyruvate Carboxylase