Mitochondrial malic enzyme: purification from bovine brain, generation of an antiserum, and immunocytochemical localization in neurons of rat brain.
To elucidate the cellular location of mitochondrial malic enzyme in brain, immunocytochemical studies were performed. For this purpose, mitochondrial malic enzyme was purified to apparent homogeneity from bovine brain and used for the immunization of rabbits. Subjecting the antiserum to affinity purification on immobilized antigen as an absorbent yielded a purified immunoreactive antibody preparation, which was characterized by probing cytosolic and mitochondrial fractions of bovine and rat brain in western blotting. As neither cross-reactivity with cytosolic malic enzyme nor immunoreactivity against other proteins could be observed, the antibody preparation was found suitable for immunocytochemistry. By using sections of perfusion-fixed rat brain, considerable resolution was achieved at the light-microscopic level. Distinct and specific staining of neurons was observed; in contrast, no staining of astrocytes and possibly unspecific staining within the nuclei of oligodendrocytes were obtained. From these data, it is concluded that mitochondrial malic enzyme is located in neurons; however, in astrocytes, the enzyme appears to be either lacking or present at a much lower level. A protective role against oxidative stress in neurons is proposed for mitochondrial malic enzyme.
Physiologisch-chemisches Institut der Universität Tübingen, Germany.
R
R
Vogel
G
G
Jennemann
J
J
Seitz
H
H
Wiesinger
B
B
Hamprecht
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D015153:Blotting, Western; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D005978:Glutathione / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000201:enzymology; D008291:Malate Dehydrogenase / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D019579:Neocortex / Q000166:cytology / Q000201:enzymology*; D009474:Neurons / Q000201:enzymology*; D018384:Oxidative Stress / Q000502:physiology; D011817:Rabbits; D051381:Rats
D000906:Antibodies; D009249:NADP; D008291:Malate Dehydrogenase; C034544:D-malate dehydrogenase (decarboxylating); D005978:Glutathione