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Differential amplification of intron-containing transcripts reveals long term potentiation-associated up-regulation of specific Pde10A phosphodiesterase splice variants

Differential amplification of intron-containing transcripts reveals long term potentiation-associated up-regulation of specific Pde10A phosphodiesterase splice variants
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 transcriptas, 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.
0021-9258
15841-15849
O'Connor, Vincent
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Genin, Alexis
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Davis, Sabrina
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Karishma, K.K.
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Doyère, Valerie
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De Zeeuw, Chris I.
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Sanger, Gareth
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Hunt, Stephen P.
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Richter-Levin, Gal
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Mallet, Jaques
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Laroche, Serge
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Bliss, T.V.P.
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French, Pim J.
df74bc1d-2be7-404f-a0a2-5edcb18c9651
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Genin, Alexis
8302af23-8cfe-4669-b5d8-dda64652abc3
Davis, Sabrina
e4c365b2-1d89-46d5-b060-291d47e3bc2d
Karishma, K.K.
742f8afe-99e0-4aad-8fe8-b8f2138c0219
Doyère, Valerie
57e5f0a7-a738-4a0b-b8b5-15ae4f7c4e92
De Zeeuw, Chris I.
477968b8-ba18-4415-9663-7918aaab571a
Sanger, Gareth
73865ecb-6c3b-46be-9175-a6168d401533
Hunt, Stephen P.
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Richter-Levin, Gal
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Mallet, Jaques
3fd455a1-de83-4cd4-9741-7cb282132126
Laroche, Serge
95d15616-f140-4257-8cdf-87c62fd8089f
Bliss, T.V.P.
a448eacf-9563-4c65-93b0-a967ac4f3d30
French, Pim J.
df74bc1d-2be7-404f-a0a2-5edcb18c9651

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. (2004) 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), 15841-15849. (doi:10.1074/jbc.M312500200).

Record type: Article

Abstract

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 transcriptas, 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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Published date: 16 April 2004

Identifiers

Local EPrints ID: 24575
URI: http://eprints.soton.ac.uk/id/eprint/24575
ISSN: 0021-9258
PURE UUID: b9b041e9-8295-438f-8dae-a02eee295592
ORCID for Vincent O'Connor: ORCID iD orcid.org/0000-0003-3185-5709

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Date deposited: 05 Apr 2006
Last modified: 29 Mar 2022 01:36

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Contributors

Author: Alexis Genin
Author: Sabrina Davis
Author: K.K. Karishma
Author: Valerie Doyère
Author: Chris I. De Zeeuw
Author: Gareth Sanger
Author: Stephen P. Hunt
Author: Gal Richter-Levin
Author: Jaques Mallet
Author: Serge Laroche
Author: T.V.P. Bliss
Author: Pim J. French

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