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J. Biol. Chem., Vol. 279, Issue 16, 15841-15849, April 16, 2004

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Differential Amplification of Intron-containing Transcripts Reveals Long Term Potentiation-associated Up-regulation of Specific Pde10A Phosphodiesterase Splice Variants^*

From the ^aSchool of Biological Sciences, University of Southampton, Southampton SO16 7PX, United Kingdom, ^dHôpital Pitié Salpetrière, UMR CNRS 9923, Laboratoire de Genetique Moleculaire de la Neurotransmission et des Processus Neurodegeneratifs, F-75013 Paris, France, ^eLaboratoire de Neurobiologie de l'Apprentisage, de la Mémoire et de la Communication, CNRS UMR 8620, Université Paris Sud, Orsay, France, ^bDivision of Neurophysiology, National Institute for Medical Research, London NW7 1AA, United Kingdom, ^gGlaxo SmithKline, Third Avenue, Harlow CM19 5AW, United Kingdom, ^hDepartment of Anatomy, University College London, London WC1E 6BT, United Kingdom, ⁱDepartment of Psychology, University of Haifa, Haifa IL-31905, Israel, and ^fDepartment of Neuroscience, Erasmus MC, 3000 DR Rotterdam, The Netherlands

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.

Received for publication, November 14, 2003 , and in revised form, January 27, 2004.

^* This work was supported by CNRS, the Medical Research Council, European Union Grant 4CT 960478 under the Framework IV program, the Hope Wessex Medical Trust, and Dutch Science Foundation Grant NWO-ALW 810-38.002. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY462091–AY462095

^c These authors contributed equally to this work.

^j To whom correspondence should be addressed. Present address: Dept. of Neurology, Erasmus MC, 3000 DR Rotterdam, The Netherlands.

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