Alpha 1-adrenoceptor subtypes involved in increased 86Rb+ influx rate and inositol 1,4,5-trisphosphate mass in adult rat cardiomyocytes

Eur J Pharmacol. 1997 Apr 4;323(2-3):269-75. doi: 10.1016/s0014-2999(97)00042-3.

Abstract

The aim of the present study was to identify the receptor subtypes involved in the alpha 1-adrenoceptor-mediated increase in 86Rb+ influx rate and in inositol 1,4,5-trisphosphate (IP3) accumulation in isolated ventricular cardiomyocytes from adult rat heart, in order to identify a possible response pattern compatible with a causative relationship. Subtype-selective receptor antagonists used were: 5-methylurapidil (alpha 1A), WB 4101 [2([2,6-dimethoxyphenoxy-ethyl]aminomethyl)-1,4-benzodioxane] (alpha 1A), chloroethylclonidine (alpha 1B) and BMY 7378 [8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4,5]dec ane-7,9-dione] (alpha 1D). The basal 86Rb+ influx rate was 0.22 +/- 0.01 ml/g protein x min. At 15 min, 5 x 10(-5) mol/l noradrenaline in the presence of 3 x 10(-5) mol/l timolol increased the 86Rb+ influx rate by 33 +/- 1%. This response was not affected by either chloroethylclonidine or BMY 7378 at concentrations up to 10(-5) mol/l. 5-Methylurapidil dose dependently inhibited the response to 5 x 10(-5) mol/l noradrenaline with a -logIC50 value of 5.27 +/- 0.12 and 5.61 +/- 0.27 in the presence and absence of 10(-5) mol/l chloroethylclonidine, respectively. WB 4101 in the presence of 10(-5) mol/l chloroethylclonidine dose dependently inhibited the response to 5 x 10(-5) mol/l noradrenaline with a -logIC50 value of 6.10 +/- 0.14. Noradrenaline in the presence of 10(-5) mol/l chloroethylclonidine dose dependently increased the 86Rb+ uptake rate with a -logEC50 value of 6.19 +/- 0.35. The basal IP3 level was 2.15 +/- 0.19 pmol/mg protein. Incubation with 10(-5) mol/l noradrenaline for 2 min increased this by 65 +/- 7% of control levels. 10(-5) mol/l chloroethylclonidine and 10(-4) mol/l 5-methylurapidil reduced the response to 27 +/- 6% and 18 +/- 9% of control level, respectively. BMY 7378 dose dependently inhibited the IP3 response at relatively high concentrations, and it was completely eliminated at 10(-5) mol/l BMY 7378. The combination of chloroethylclonidine and 5-methylurapidil or 3 x 10(-6) mol/l prazosin alone completely abolished the hormone-induced effect. We conclude that whereas the alpha 1-adrenoceptor-stimulated increase in 86Rb+ influx rate is mediated via the alpha 1A-adrenoceptor subtype only, both alpha 1A- and alpha 1B-adrenoceptor subtypes are involved in the increase in IP3 mass. Furthermore, a contribution from the alpha 1D-adrenoceptor in the IP3 response cannot be excluded. Thus there does not appear to be a simple causative relationship between an increase in 86Rb+ influx rate and an increase in IP3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenergic alpha-1 Receptor Antagonists*
  • Adrenergic alpha-2 Receptor Antagonists
  • Adrenergic alpha-Antagonists / pharmacology
  • Animals
  • Cells, Cultured
  • Clonidine / analogs & derivatives
  • Clonidine / pharmacology
  • Dioxanes / pharmacology
  • Heart Ventricles / metabolism
  • Inositol 1,4,5-Trisphosphate / metabolism*
  • Myocardium / cytology
  • Myocardium / metabolism*
  • Piperazines / pharmacology
  • Rats
  • Receptors, Adrenergic, alpha-1
  • Receptors, Adrenergic, alpha-2
  • Rubidium Radioisotopes / metabolism*

Substances

  • Adra1a protein, rat
  • Adra1d protein, rat
  • Adra2a protein, rat
  • Adrenergic alpha-1 Receptor Antagonists
  • Adrenergic alpha-2 Receptor Antagonists
  • Adrenergic alpha-Antagonists
  • Dioxanes
  • Piperazines
  • Receptors, Adrenergic, alpha-1
  • Receptors, Adrenergic, alpha-2
  • Rubidium Radioisotopes
  • 5-methylurapidil
  • chlorethylclonidine
  • Inositol 1,4,5-Trisphosphate
  • (2-(2',6'-dimethoxy)phenoxyethylamino)methylbenzo-1,4-dioxane
  • BMY 7378
  • Clonidine