Differential expression of protein phosphatase 1 isoforms in mammalian brain.
Rat cDNAs encoding neuronal isoforms of protein phosphatase 1 (PP1) were isolated and their primary structures elucidated. The derived amino acid sequences allowed us to design synthetic C-terminal peptides that were used to raise antibodies. Isoform-specific anti-peptide antibodies against PP1 alpha and PP1 gamma 1 were used to investigate the tissue distribution of PP1 isoforms by immunoblotting. Both isoforms were ubiquitously expressed in mammalian tissues, with the highest levels being observed in brain. Of all neuronal tissues examined, PP1 alpha and PP1 gamma 1 were found to be most abundantly expressed in the striatum. Lesion experiments with kainic acid indicated that both the alpha and the gamma 1 isoforms of protein phosphatase 1 were relatively enriched in the medium-size spiny neurons of the striatum. "In situ" hybridization to rat brain slices using highly sensitive riboprobes also showed PP1 alpha, PP1 beta, and PP1 gamma 1 to be widely expressed in mammalian brain. However, some interesting differences were observed. For example, PP1 alpha and PP1 gamma 1 were found to be expressed in the striatum, where DARPP-32 (dopamine- and cAMP-regulated phosphoprotein, M(r) = 32,000 Da) is also known to be highly expressed. PP1 beta appeared to be relatively less abundant in the same cells, as judged both by "in situ" hybridization and by the apparent absence of PP1 beta clones from the striatal cDNA libraries used.
da Cruz e Silva
E F
EF
Rockefeller University, New York, New York 10021, USA.
Fox
C A
CA
Ouimet
C C
CC
Gustafson
E
E
Watson
S J
SJ
Greengard
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D001921:Brain / Q000166:cytology / Q000201:enzymology*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D015870:Gene Expression; D015723:Gene Library; D015151:Immunoblotting; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000096:biosynthesis*; D008297:Male; D008969:Molecular Sequence Data; D009474:Neurons / Q000201:enzymology*; D009928:Organ Specificity; D010749:Phosphoprotein Phosphatases / Q000096:biosynthesis*; D010761:Phosphorus Radioisotopes; D016548:Prosencephalon / Q000166:cytology / Q000201:enzymology; D054645:Protein Phosphatase 1; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D007527:Isoenzymes; D010761:Phosphorus Radioisotopes; D015347:RNA Probes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1