Inhibitors of protein phosphatase-2A: topography and subcellular localization.

The mRNA and protein expressions of I1(PP2A) and I2(PP2A), the two inhibitors of protein phosphatase 2A (PP2A) were investigated in adult rat brain. The rat brain and human brain inhibitors showed similar molecular weights by Western blots. The cDNA probes for human brain I1(PP2A) and I2(PP2A) readily hybridized with the corresponding mRNAs of rat brain inhibitors in Northern blots. We detected 3.7 and 2.1 kb transcripts of I1(PP2A) and 2.9 and 2.0 kb transcripts of I2(PP2A) in rat brain. In situ hybridization revealed that the mRNAs of the two inhibitors were mainly localized in neurons. Strong expression of both I1(PP2A) and I2(PP2A) mRNAs were observed in the olfactory bulb, hippocampal pyramidal and dentate granule cell layers, and cerebellar Purkinje cell, granular and molecular layers. Moderate expression of I1(PP2A) and I2(PP2A) mRNAs were observed in the cerebral cortex, caudate putamen, thalamus, hypothalamus, amygdala and pontine nucleus. The expression of I1(PP2A) and I2(PP2A) and as well as of PP-2A was also investigated by immunohistochemistry using antibodies to each protein. The distribution patterns of the two inhibitor proteins were similar to those of their corresponding mRNAs and to the expression of PP-2A. While PP-2A was localized to neuronal perikarya, I1(PP2A) was observed both in the neuronal cytoplasm and the nucleus. I2(PP2A) had mainly nuclear localization but it could also be seen in the neuronal cytoplasm. All three proteins were also expressed in the neuropil. These studies suggest that PP-2A activity is probably regulated by I1(PP2A) and I2(PP2A) in the adult mammalian central nervous system, and that these inhibitors are conserved between rat and human brains.