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Immunocytochemical Examination of Neural Rat and Mouse Primary Cultures Using Monoclonal Antibodies Raised Against Pyruvate Carboxylase
Author(s) -
Cesar Marija,
Hamprecht Bernd
Publication year - 1995
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 - 10.1046/j.1471-4159.1995.64052312.x
Subject(s) - enolase , glial fibrillary acidic protein , colocalization , biology , monoclonal antibody , microbiology and biotechnology , pyruvate carboxylase , neuron , immunocytochemistry , neuroglia , astrocyte , cell culture , antibody , biochemistry , immunohistochemistry , enzyme , central nervous system , immunology , neuroscience , endocrinology , genetics
Pyruvate carboxylase (EC 6.4.1.1; PC) catalyzes the formation of oxaloacetate by energy‐dependent fixation of CO 2 to pyruvate. The aim of the present work was to generate antibodies against PC and use them to localize PC in the cells of astroglia‐rich and neuron‐rich primary cultures derived from the brains of rats and mice. Mouse monoclonal antibodies raised against the enzyme were shown to be monospecific as indicated by immunoblotting. The staining of the cells for PC appeared in grains. These represent mitochondria, as PC is known as a mitochondrial enzyme. Immunocytochemical examination of astroglia‐rich primary cultures of rat or mouse brain cells revealed a colocalization of PC with the astroglial marker glial fibrillary acidic protein (GFAP) in many cells. However, there were GFAP‐positive cells showing no specific staining for PC, and vice versa. Also, in neuron‐rich primary cultures PC was found only in the ∼10% GFAP‐expressing astroglial cells contaminating the neuron‐rich primary culture, whereas it was absent from the neurons identified by antibodies against neuron‐specific enolase. These results suggest that PC is predominantly an astroglial enzyme and that astroglial cells play an important role in the intermediary and the energy metabolism of the brain.