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

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)949-958
Number of pages10
JournalLife Sciences
Volume52
Issue number11
DOIs
StatePublished - 1993
Externally publishedYes

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Receptor, Serotonin, 5-HT1A
HeLa Cells
Rats
Cells
Cell Line
Hippocampus
Genes
Serotonin 5-HT1 Receptor Agonists
Buspirone
Spiperone
Cell membranes
Guanosine Triphosphate
GTP-Binding Proteins
Serotonin
Binding Sites
Cell Membrane
Pharmaceutical Preparations
Proteins

ASJC Scopus subject areas

  • Pharmacology

Cite this

Fujiwara, Y., Tomita, H., Hikiji, M., Kashihara, K., Otsuki, S., Ohnuki, T., ... Yamamura, H. I. (1993). Characterization of a cloned rat serotonin 5-HT1A receptor expressed in the HeLa cell line. Life Sciences, 52(11), 949-958. https://doi.org/10.1016/0024-3205(93)90530-G

Characterization of a cloned rat serotonin 5-HT1A receptor expressed in the HeLa cell line. / Fujiwara, Yutaka; Tomita, Hiroaki; Hikiji, Mitsuru; Kashihara, Kenichi; Otsuki, Saburo; Ohnuki, Tetsuo; Hamagishi, Yasutaro; Oki, Toshikazu; Sora, Ichiro; Roeske, William R.; Yamamura, Henry I.

In: Life Sciences, Vol. 52, No. 11, 1993, p. 949-958.

Research output: Contribution to journalArticle

Fujiwara, Y, Tomita, H, Hikiji, M, Kashihara, K, Otsuki, S, Ohnuki, T, Hamagishi, Y, Oki, T, Sora, I, Roeske, WR & Yamamura, HI 1993, 'Characterization of a cloned rat serotonin 5-HT1A receptor expressed in the HeLa cell line', Life Sciences, vol. 52, no. 11, pp. 949-958. https://doi.org/10.1016/0024-3205(93)90530-G
Fujiwara, Yutaka ; Tomita, Hiroaki ; Hikiji, Mitsuru ; Kashihara, Kenichi ; Otsuki, Saburo ; Ohnuki, Tetsuo ; Hamagishi, Yasutaro ; Oki, Toshikazu ; Sora, Ichiro ; Roeske, William R. ; Yamamura, Henry I. / Characterization of a cloned rat serotonin 5-HT1A receptor expressed in the HeLa cell line. In: Life Sciences. 1993 ; Vol. 52, No. 11. pp. 949-958.
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AU - Tomita, Hiroaki

AU - Hikiji, Mitsuru

AU - Kashihara, Kenichi

AU - Otsuki, Saburo

AU - Ohnuki, Tetsuo

AU - Hamagishi, Yasutaro

AU - Oki, Toshikazu

AU - Sora, Ichiro

AU - Roeske, William R.

AU - Yamamura, Henry I.

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