Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 317, Issue 4, pp 294–301 | Cite as

Desensitization of the β-adrenoceptor-adenylate cyclase system of immature erythrocytes by in-vivo treatment of rats with isoprenaline

  • G. Wiemer
  • G. Kaiser
  • J. Dietz
  • M. Reinhardt
  • A. Wellstein
  • D. Palm


Rats pretreated with 1-acetyl-2-phenylhydrazide, to induce reticulocytosis, were treated with (±)isoprenaline (4×30 mg/kg in 6 h intervals within 24 h before blood sampling) in order to desensitize the β-adrenoceptor-adenylate cyclase-system of circulating red cells.

  1. 1.

    In membrane preparations Vmax-values of (-)isoprenaline-sensitive adenylate cyclase activity declined by about 50% without significant alterations of the apparent Km-values. Basal activity, as well as enzyme activity stimulated maximally by guanylyl-imidodiphosphate and by fluoride were also decreased to the same extent.

  2. 2.

    In intact cells, maximal synthesis of cAMP stimulated by 10−5 M (-)isoprenaline was decreased by about 50% compared to cell suspensions from control animals in the presence of Ro 20-1724 or papaverine, inhibitors of phosphodiesterase. Without inhibition of phosphodiesterase, (-)isoprenaline stimulated cAMP synthesis in cell suspensions from desensitized animals exceeded that from control animals more than 10-fold. A 20–30% decrease of phosphodiesterase activity, measured in membrane and cytoplasmic fractions explains this unexpected result.

  3. 3.

    In membrane preparations from untreated animals, Bmax-values for the antagonist ligand [3H] dihydroalprenolol ([3H] DHA) were (0.98±0.16 pmoles/mg protein (n=7). For the agonist-ligand [3H] hydroxybenzylisoprenaline ([3H]-HBI) only 0.23±0.024 pmoles/mg protein (n=12) were obtained. After pretreatment of the animals with (±)isoprenaline, the Bmax-value for [3H] HBI was decreased by about 50%; whereas, that for [3H] DHA was decreased only by about 10%. No change in KD-values for both ligands occurred.

  4. 4.

    It is proposed that β-adrenergic desensitization in immature red cells, i.e. the decrease of (-)isoprenalinesensitive adenylate cyclase activity, results from loss of high affinity β-adrenoceptor sites. This might be induced by uncoupling the adrenoceptors from the nucleotide binding protein. A functional impairment or loss of this regulatory protein may be responsible for the apparent loss of adenylate cyclase activity. Since [3H] HBI, in contrast to [3H] DHA, labels predominantly β-adrenoceptors in the high affinity state it is a more sensitive marker to detect alterations at the adrenoceptor level following desensitization.


Key words

Desensitization β-Adrenoceptors Adenylate cyclase Agonist binding Rat reticulocytes 


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Copyright information

© Springer-Verlag 1981

Authors and Affiliations

  • G. Wiemer
    • 1
  • G. Kaiser
    • 1
  • J. Dietz
    • 1
  • M. Reinhardt
    • 1
  • A. Wellstein
    • 1
  • D. Palm
    • 1
  1. 1.Zentrum der PharmakologieKlinikum der Johann Wolfgang Goethe-UniversitätFrankfurt/MainGermany

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