Differential amplification of intron-containing transcripts reveals long term potentiation-associated up-regulation of specific Pde10A phosphodiesterase splice variants.
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.
O'Connor
Vincent
V
School of Biological Sciences, University of Southampton, Southampton SO16 7PX, United Kingdom.
Genin
Alexis
A
Davis
Sabrina
S
Karishma
K K
KK
Doyère
Valerie
V
De Zeeuw
Chris I
CI
Sanger
Gareth
G
Hunt
Stephen P
SP
Richter-Levin
Gal
G
Mallet
Jacques
J
Laroche
Serge
S
Bliss
T V P
TV
French
Pim J
PJ
D000818:Animals; D006801:Humans; D007438:Introns; D007527:Isoenzymes / Q000235:genetics; D017774:Long-Term Potentiation / Q000235:genetics*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009473:Neuronal Plasticity / Q000235:genetics; D010727:Phosphoric Diester Hydrolases / Q000235:genetics*; D012326:RNA Splicing; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation / Q000235:genetics
D007527:Isoenzymes; C119843:PDE10A protein, human; C492284:Pde10a protein, mouse; D010727:Phosphoric Diester Hydrolases