Characterization of ai-adrenoceptors expressed in a novel vascular smooth muscle cell line cloned from p53 knockout mice, P53LMAC01 (AC01) cells

Kazuhiro Ohmi, Hitomi Shinoura, Yasuhisa Nakayama, Nobuhito Goda, Gozoh Tsujimoto

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We pharmacologically studied the a,-adrenoceptor (AR) subtype(s) involved in receptormediated signalling in a novel vascular smooth muscle cell line cloned from p53 knockout mice, P53LMAC01 (AC01) cells. 2 Radioligand binding studies with [125I]-HEAT showed the existence of a homogeneous population of binding site with an affinity (K, value) of 0.4 nM and a maximum number of binding .sites (Bmm) of lOOfmolmg"1 protein. Catecholamines competed for [125I]-HEAT binding stereospecifically and with the characteristic oi-AR potency series. 3 Displacement curves for BMY-7378 and KMD-3213 best fitted a one-site model with a pKj value (-logio (equilibrium inhibition constant)) of 6.06 and 7.07, respectively. 4 Reverse transcription-polymerase chain reaction (RT-PCR) assay detected m- and aiD-AR, but not IA-AR transcript. 5 Chlorethylclonidine (CEC) treatment nearly abolished (-)noradrenaline (NA) (10 /(M)-induced inositol[l,4,5]trisphosphate (IP3) production, and BMY-7378 inhibited the response with a K, value of 0.3 nM, which value was similar to that obtained in the cells expressing 1D-AR. In both AC01 cells and cells expressing ociD-AR, BMY-7378 protected i-ARs from CEC alkylation while it had little protective effect on CEC alkylation and NA-induced IP3 production in cells expressing a,B-AR. 6 The results indicate that AC01 cells contain predominantly a]B-ARs and a small population of a1D-ARs; however, phosphoinositide (PI)/Ca2 + signalling is mainly mediated through the minor population of a)D-ARs, rather than the iB-ARs.

Original languageEnglish
Pages (from-to)756-762
Number of pages7
JournalBritish Journal of Pharmacology
Volume127
Issue number3
Publication statusPublished - 1999
Externally publishedYes

Fingerprint

Vascular Smooth Muscle
Knockout Mice
Adrenergic Receptors
Smooth Muscle Myocytes
Cell Line
Alkylation
Norepinephrine
Population
Inositol
Phosphatidylinositols
Reverse Transcription
Catecholamines
Binding Sites
Polymerase Chain Reaction
chlorethylclonidine
BMY 7378
Proteins

Keywords

  • Aradrenoceptor
  • Noradrenaline
  • Vascular smooth muscle cells

ASJC Scopus subject areas

  • Pharmacology

Cite this

Characterization of ai-adrenoceptors expressed in a novel vascular smooth muscle cell line cloned from p53 knockout mice, P53LMAC01 (AC01) cells. / Ohmi, Kazuhiro; Shinoura, Hitomi; Nakayama, Yasuhisa; Goda, Nobuhito; Tsujimoto, Gozoh.

In: British Journal of Pharmacology, Vol. 127, No. 3, 1999, p. 756-762.

Research output: Contribution to journalArticle

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abstract = "We pharmacologically studied the a,-adrenoceptor (AR) subtype(s) involved in receptormediated signalling in a novel vascular smooth muscle cell line cloned from p53 knockout mice, P53LMAC01 (AC01) cells. 2 Radioligand binding studies with [125I]-HEAT showed the existence of a homogeneous population of binding site with an affinity (K, value) of 0.4 nM and a maximum number of binding .sites (Bmm) of lOOfmolmg{"}1 protein. Catecholamines competed for [125I]-HEAT binding stereospecifically and with the characteristic oi-AR potency series. 3 Displacement curves for BMY-7378 and KMD-3213 best fitted a one-site model with a pKj value (-logio (equilibrium inhibition constant)) of 6.06 and 7.07, respectively. 4 Reverse transcription-polymerase chain reaction (RT-PCR) assay detected m- and aiD-AR, but not IA-AR transcript. 5 Chlorethylclonidine (CEC) treatment nearly abolished (-)noradrenaline (NA) (10 /(M)-induced inositol[l,4,5]trisphosphate (IP3) production, and BMY-7378 inhibited the response with a K, value of 0.3 nM, which value was similar to that obtained in the cells expressing 1D-AR. In both AC01 cells and cells expressing ociD-AR, BMY-7378 protected i-ARs from CEC alkylation while it had little protective effect on CEC alkylation and NA-induced IP3 production in cells expressing a,B-AR. 6 The results indicate that AC01 cells contain predominantly a]B-ARs and a small population of a1D-ARs; however, phosphoinositide (PI)/Ca2 + signalling is mainly mediated through the minor population of a)D-ARs, rather than the iB-ARs.",
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AU - Ohmi, Kazuhiro

AU - Shinoura, Hitomi

AU - Nakayama, Yasuhisa

AU - Goda, Nobuhito

AU - Tsujimoto, Gozoh

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N2 - We pharmacologically studied the a,-adrenoceptor (AR) subtype(s) involved in receptormediated signalling in a novel vascular smooth muscle cell line cloned from p53 knockout mice, P53LMAC01 (AC01) cells. 2 Radioligand binding studies with [125I]-HEAT showed the existence of a homogeneous population of binding site with an affinity (K, value) of 0.4 nM and a maximum number of binding .sites (Bmm) of lOOfmolmg"1 protein. Catecholamines competed for [125I]-HEAT binding stereospecifically and with the characteristic oi-AR potency series. 3 Displacement curves for BMY-7378 and KMD-3213 best fitted a one-site model with a pKj value (-logio (equilibrium inhibition constant)) of 6.06 and 7.07, respectively. 4 Reverse transcription-polymerase chain reaction (RT-PCR) assay detected m- and aiD-AR, but not IA-AR transcript. 5 Chlorethylclonidine (CEC) treatment nearly abolished (-)noradrenaline (NA) (10 /(M)-induced inositol[l,4,5]trisphosphate (IP3) production, and BMY-7378 inhibited the response with a K, value of 0.3 nM, which value was similar to that obtained in the cells expressing 1D-AR. In both AC01 cells and cells expressing ociD-AR, BMY-7378 protected i-ARs from CEC alkylation while it had little protective effect on CEC alkylation and NA-induced IP3 production in cells expressing a,B-AR. 6 The results indicate that AC01 cells contain predominantly a]B-ARs and a small population of a1D-ARs; however, phosphoinositide (PI)/Ca2 + signalling is mainly mediated through the minor population of a)D-ARs, rather than the iB-ARs.

AB - We pharmacologically studied the a,-adrenoceptor (AR) subtype(s) involved in receptormediated signalling in a novel vascular smooth muscle cell line cloned from p53 knockout mice, P53LMAC01 (AC01) cells. 2 Radioligand binding studies with [125I]-HEAT showed the existence of a homogeneous population of binding site with an affinity (K, value) of 0.4 nM and a maximum number of binding .sites (Bmm) of lOOfmolmg"1 protein. Catecholamines competed for [125I]-HEAT binding stereospecifically and with the characteristic oi-AR potency series. 3 Displacement curves for BMY-7378 and KMD-3213 best fitted a one-site model with a pKj value (-logio (equilibrium inhibition constant)) of 6.06 and 7.07, respectively. 4 Reverse transcription-polymerase chain reaction (RT-PCR) assay detected m- and aiD-AR, but not IA-AR transcript. 5 Chlorethylclonidine (CEC) treatment nearly abolished (-)noradrenaline (NA) (10 /(M)-induced inositol[l,4,5]trisphosphate (IP3) production, and BMY-7378 inhibited the response with a K, value of 0.3 nM, which value was similar to that obtained in the cells expressing 1D-AR. In both AC01 cells and cells expressing ociD-AR, BMY-7378 protected i-ARs from CEC alkylation while it had little protective effect on CEC alkylation and NA-induced IP3 production in cells expressing a,B-AR. 6 The results indicate that AC01 cells contain predominantly a]B-ARs and a small population of a1D-ARs; however, phosphoinositide (PI)/Ca2 + signalling is mainly mediated through the minor population of a)D-ARs, rather than the iB-ARs.

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KW - Noradrenaline

KW - Vascular smooth muscle cells

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