O'Connor, Vincent, Genin, Alexis, Davis, Sabrina, Karishma, K.K., Doyère, Valerie, De Zeeuw, Chris I., Sanger, Gareth, Hunt, Stephen P., Richter-Levin, Gal, Mallet, Jaques, Laroche, Serge, Bliss, T.V.P. and French, Pim J.
Differential amplification of intron-containing transcripts reveals long term potentiation-associated up-regulation of specific Pde10A phosphodiesterase splice variants
The Journal of Biological Chemistry, 279, (16), . (doi:10.1074/jbc.M312500200).
We employed differential display of expressed mRNAs
(Liang, P., and Pardee, A. B. (1992) Science 257, 967–971)
to identify genes up-regulated after long term potentiation
(LTP) induction in the hippocampus of awake adult
rats. In situ hybridization confirmed the differential expression
of five independently amplified clones representing
two distinct transcripts, cl13/19/90 and cl95/96.
Neither cl13/19/90 nor cl95/96 showed significant sequence
homology to known transcripts (mRNA or expressed
sequence tag) or to the mouse or human genome.
However, comparison with the rat genome
revealed that they are localized to a predicted intron of
the phosphodiesterase Pde10A gene. cl13/19/90 and
cl95/96 are likely to be part of the Pde10A primary transcript
as, using reverse transcriptase-PCR, we could
specifically amplify distinct introns of the Pde10A primary
transcript, and in situ hybridization demonstrated
that a subset of Pde10A splice variants are also up-regulated
after LTP induction. These results indicate that
amplification of a primary transcript can faithfully report
gene activity and that differential display can be
used to identify differential expression of RNA species
other than mRNA. In transiently transfected Cos7 cells,
Pde10A3 reduces the atrial natriuretic peptide-induced
elevation in cGMP levels without affecting basal cGMP
levels. This cellular function of LTP-associated Pde10A
transcripts argues for a role of the cGMP/cGMP-dependent
kinase pathway in long term synaptic plasticity.
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