Glutamate carboxypeptidase II is expressed by astrocytes in the adult rat nervous system.
The enzyme glutamate carboxypeptidase II (GCP II) has been cloned from rat brain and human prostate. This enzyme, which catabolizes the neuropeptide N-acetylaspartylglutamate, has also been known as N-acetylated alpha-linked acidic dipeptidase (NAALADase), and is identical to the prostate-specific membrane antigen and to the jejunal folylpoly-gamma-glutamate carboxypeptidase. The goals of the present study were to elucidate the cell specificity and regional pattern of GCP II expression in the rat nervous system by using Northern blots and enzymatic assays of brain and subfractionated primary neuronal and glial cultures together with in situ hybridization histochemistry (ISHH) in sections of adult rat tissue. GCP II activity was assayed in astrocyte cultures (4.4 pmol/mg protein per minute), neuronal-glial cocultures (2.5 pmol/mg protein per minute) and neuron-enriched cultures (0.38 pmol/mg protein per minute), with the activity in each preparation correlating to its astrocytic content (r = 0.99). No activity was detected in cultured oligodendrocytes or microglia. Northern blots probed with a GCP II cDNA detected mRNAs exclusively in activity-positive cell preparations. ISHH results show that GCP II is expressed by virtually all astrocytes, by Bergmann glial cells in cerebellum, by Müller cells in retina and by the satellite cells in dorsal root ganglia. Astrocytes in select groups of nuclei (e.g., habenula, supraoptic nucleus, pontine nucleus) contained pronounced levels of GCP II message. The data of the present study suggest that GCP II is expressed in the adult rat nervous system exclusively in astrocytic glial cells.
Berger
U V
UV
Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
Luthi-Carter
R
R
Passani
L A
LA
Elkabes
S
S
Black
I
I
Konradi
C
C
Coyle
J T
JT
D000818:Animals; D000954:Antigens, Surface; D001253:Astrocytes / Q000201:enzymology*; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology*; D002268:Carboxypeptidases / Q000235:genetics*; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D043425:Glutamate Carboxypeptidase II; D006801:Humans; D008297:Male; D009457:Neuroglia / Q000201:enzymology*; D009474:Neurons / Q000201:enzymology*; D009928:Organ Specificity; D017378:Quinolinic Acid / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000201:enzymology*; D014158:Transcription, Genetic
D000954:Antigens, Surface; D012333:RNA, Messenger; D002268:Carboxypeptidases; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II; D017378:Quinolinic Acid