Molecular cloning and identification of a rabbit saphenous vein 5-ht1Dβ receptor gene
✍ Scribed by Thierry Wurch; Claudie Cathala; Christiane Palmier; Jean-Pierre Valentin; Gareth W. John; Francis C. Colpaert; Petrus J. Pauwels
- Publisher
- John Wiley and Sons
- Year
- 1996
- Tongue
- English
- Weight
- 571 KB
- Volume
- 18
- Category
- Article
- ISSN
- 0893-6609
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✦ Synopsis
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-HT -like receptor subtype in this tissue, three sets of oligonucleotide primers were designed according to t fi e human 5-HT1 gene sequence for use in reverse transcription-polymerase chain reaction (RT-PCR). Amp i cation 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 S-HTID~, rat 5-HT B and mouse ~-H T ~B receptor subtypes. A Thr-355 was found in trans-membrane domain VII as 1 or the human 5 -H T l ~p 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 2 Zolmitriptan > MK-462 2 Sumatriptan > (+)-8-OH-DPAT > CP 93,129. This binding profile together with its amino acid sequence indicate that this rabbit gene encodes a 5 -h t l ~n receptor.