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1 Movetis NV, Turnhout, Antwerpen, Belgium
2 Tibotec bvba, Mechelen, Antwerpen, Belgium
3 Department of Functional Genomics, Johnson & Johnson Pharmaceutical Research and Development, Beerse, Antwerpen, Belgium
4 Heymans Institute of Pharmacology, Ghent University, Gent, Oost-Vlaanderen, Belgium
* To whom correspondence should be addressed. E-mail: joris.demaeyer{at}movetis.com.
5-HT4 receptors are present in human and porcine atrial myocytes while they are absent from the heart of small laboratory animals. The pig is therefore the only available non-primate animal model to study cardiac 5-HT4 receptor function under physiological conditions. While several human splice variants of the 5-HT4 receptor have been described, the splicing behaviour of this receptor in porcine tissue is currently unknown. Here we report on the identification of nine novel C-terminal splice variants of the porcine 5-HT4 receptor, which were named 5-HT4(b2, j, k, l, m, o, p, q, r). The internal h-variant was found in combination with several C-terminal exons. In addition, splice variants were found that comprised duplicated exons fused to the common region of the 5-HT4 receptor, thereby providing evidence for a duplication of the porcine HTR4 gene. One of these variants putatively encoded a nine transmembrane-spanning domain homofusion receptor, 5-HT4(9TM); also the other variants with a duplicated region might translate into functional, transcriptionally fused dimeric 5-HT4 receptor variants. The elucidation of the genomic context confirmed that the variants were not genomic artefacts but originated from alternative splicing. This was further corroborated by a functional analysis of the variants 5-HT4(a), 5-HT4(r) and 5-HT4(9TM). To our knowledge, our data are the first to report on a functional GPCR with more than 7 predicted transmembrane domains. These findings urge for caution when interpreting data on 5-HT4 receptor-related pharmacology obtained in the pig; validation at the molecular level might be needed before extrapolating results to human.
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