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 language | English |
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Pages (from-to) | 155-162 |
Number of pages | 8 |
Journal | Neuroscience Research Communications |
Volume | 18 |
Issue number | 3 |
DOIs | |
State | Published - 1996 |
Externally published | Yes |
Keywords
- 5-HT receptor
- Binding profile
- Gene cloning
- RT-PCR
- Rabbit saphenous vein
- Transient expression