Distribution of glutamic acid decarboxylase (Mr 67,000) in the basal ganglia of the rat: an immunohistochemical study with a selective cDNA-generated polyclonal antibody.
Distinct isoforms of glutamic acid decarboxylase, the synthetic enzyme for GABA, exist in brain. Their distribution at the cellular level is not known, because previous studies have been confounded by the lack of monospecificity of available antibodies. We have examined the distribution of glutamic acid decarboxylase (Mr 67,000; GAD67) in the basal ganglia of the rat with a polyclonal antibody generated against the protein expressed in bacteria transformed with the corresponding cDNA. This antibody, which is directed against a portion of GAD67 non homologous to other known glutamic acid decarboxylase isoforms, selectively recognizes GAD67 on western blots. We show that GAD67 is present to various degree in all types of GABAergic neurons previously described in these regions. In contrast with results obtained with non-selective antibodies for glutamic acid decarboxylase, GAD67-positive neuronal cell bodies were readily detected in sections of the striatum, pallidum and substantia nigra in the absence of colchicine treatment. Modifications in the immunohistochemical procedure favoured staining of glutamic acid decarboxylase-positive fibres with the same antibody, indicating that GAD67 is also present in axon terminals of GABAergic neurons. The results suggest that GAD67 may be involved in GABA synthesis in both cell bodies and axon terminals of all GABAergic neurons of the basal ganglia, but is particularly abundant or accessible in their cell bodies.
Gonzales
C
C
Department of Pharmacology, University of Pennsylvania, Philadelphia 19104.
Kaufman
D L
DL
Tobin
A J
AJ
Chesselet
M F
MF
D000818:Animals; D001479:Basal Ganglia / Q000201:enzymology*; D003342:Corpus Striatum / Q000201:enzymology; D004247:DNA / Q000235:genetics; D005917:Globus Pallidus / Q000201:enzymology; D005968:Glutamate Decarboxylase / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D007150:Immunohistochemistry; D008297:Male; D008970:Molecular Weight; D009412:Nerve Fibers / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins / Q000276:immunology; D013378:Substantia Nigra / Q000201:enzymology; D014169:Transformation, Bacterial; D005680:gamma-Aminobutyric Acid / Q000096:biosynthesis
D011994:Recombinant Proteins; D005680:gamma-Aminobutyric Acid; D004247:DNA; D005968:Glutamate Decarboxylase