Molecular cloning and identification of a rabbit saphenous vein 5-ht1Dβ receptor gene

Thierry Wurch, Claudie Cathala, Christiane Palmier, Jean Pierre Valentin, Gareth W. John, Francis C. Colpaert, Petrus J. Pauwels

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

The molecular identity of the serotonin (5-HT) receptor subtypes that mediate contraction in the rabbit isolated saphenous vein remains unclear. In order to identify a 5-HT1-like receptor subtype in this tissue, three sets of oligonucleotide primers were designed according to the human 5-HT1Dβ receptor gene sequence for use in reverse transcription-polymerase chain reaction (RT-PCR). Amplification of specific PCR-products was obtained with rabbit saphenous vein total RNA reverse-transcribed into single-stranded cDNA.The PCR-amplified products were used to screen a rabbit genomic library. Sequencing of PCR-products and of a library clone revealed an open reading frame of 1173 base pairs. The deduced amino acid sequence is 91 to 93 % homologous to the human 5-HT1Dβ, rat 5-HT1B and mouse 5-HT1B receptor subtypes. A Thr-355 was found in trans-membrane domain VII as for the human 5-HT1Dβ receptor. Transient expression of this rabbit saphenous vein gene in Cos-7 cells yielded the following receptor binding profile : 5-Carboxamidotryptamine > 5-HT > Methiothepin > Naratriptan ≥ Zolmitriptan > MK-462 ≥ Sumatriptan > (+)-8-OH-DPAT > CP 93,129. This binding profile together with its amino acid sequence indicate that this rabbit gene encodes a 5-ht1Dβ receptor.

Original languageEnglish
Pages (from-to)155-162
Number of pages8
JournalNeuroscience Research Communications
Volume18
Issue number3
DOIs
StatePublished - 1996
Externally publishedYes

Keywords

  • 5-HT receptor
  • Binding profile
  • Gene cloning
  • RT-PCR
  • Rabbit saphenous vein
  • Transient expression

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