Characterization of a cloned rat serotonin 5-HT1A receptor expressed in the HeLa cell line

Yutaka Fujiwara, Hiroaki Tomita, Mitsuru Hikiji, Kenichi Kashihara, Saburo Otsuki, Tetsuo Ohnuki, Yasutaro Hamagishi, Toshikazu Oki, Ichiro Sora, William R. Roeske, Henry I. Yamamura

研究成果: Article査読

15 被引用数 (Scopus)

抄録

We have previously isolated the rat serotonin (5-HT)1A receptor gene (G21Y2) and now report the expression and characterization of this receptor. The BamHl/Xbal fragment of this gene was cloned into Rc/RSV and stably transfected into HeLa cells by the calcium phosphate method. For determination of specific 5-HT1A receptor binding, [3H]8OH-DPAT was used as the radioligand and incubated with HeLa cell membranes. The cells expressed specific and saturable binding of [3H]8OH-DPAT with a Kd value of 0.3 nM and a Bmax value of 2 pmol/mg protein. GTP (50 μM) added to the incubation mixture increased the Kd value to 3 nM indicating that the expressed receptor is coupled to a G protein. The specific binding was inhibited by selective 5-HT1A partial agonists, such as buspirone, ipsapirone, gepirone, tandospirone, zalospirone and SUN8399 with Ki values of 1-30nM, whereas other neurotropic drugs except for spiperone (Ki=46 nM) and nemonapride (Ki=2.3 nM) were effective only at concentrations of more than 100 μM. The potencies of these compounds to inhibit [3H]8OH-DPAT from its specific binding sites were similar to their affinities determined in rat hippocampus binding studies. These data suggest that the expressed receptor is a 5-HT1A-type similar to 5-HT1A receptors in the rat hippocampus.

本文言語English
ページ(範囲)949-958
ページ数10
ジャーナルLife Sciences
52
11
DOI
出版ステータスPublished - 1993
外部発表はい

ASJC Scopus subject areas

  • 生化学、遺伝学、分子生物学(全般)
  • 薬理学、毒性学および薬学(全般)

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