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G-protein-coupled receptors in normal human erythroid progenitor cells

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Abstract

Human erythroid progenitor cells were isolated from peripheral blood of healthy donors and amplified in a suspension culture system using recombinant growth factors (stem cell factor, interleukin-3, granulocyte-macrophage colony-stimulating factor and erythropoietin) as well as conditioned medium from a human bone marrow stroma cell line to support cell proliferation. After 6–8 days of culture, the cell population consisted mainly of erythroid colony-forming cells (burst-forming units, BFU-Es and colony-forming units, CFU-Es). In these cells, we studied ligand-induced changes in intracellular Ca 2+ concentration ([Ca2+]i) and cAMP formation as the primary effector systems of guanine nucleotide-binding protein (G protein)-coupled receptors. The results confirmed the functional expression of receptors for adenosine (type A2B), prostaglandin Et and isoprenaline (\-adrenoceptor), all of which stimulated adenylyl cyclase, as well as for ADP (purinoceptor types P2T and P2U), platelet-activating factor and thrombin all of which caused a transient increase in [Ca2+]j. The efficacy of adenosine and prostaglandin El in stimulating cAMP formation was more than 5 times higher than that of isoprenaline, suggesting a low \-adrenoceptor density. The response to adenosine and isoprenaline decreased by 80 and 55% respectively during maturation into the proerythroblast stage. Similarly, thapsigargin-sensitive intracellular Ca 2+ stores and ligand-induced Ca 2+ release declined by about 60% during the CFU-E-to-erythroblast transition. The overall functional expression pattern of G protein-coupled receptors differed from that in human erythroleukaemia cell lines or from that in platelets. Primary culture systems for nontransformed cells, such as the one presented here, thus will be indispensable for the study of the functional role of G protein-dependent signalling during haematopoiesis.

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Authors and Affiliations

  1. Department of Pharmacology, Universität Bern, Friedbühlstrasse 49, CH-3010, Bern, Switzerland

    H. Porzig, R. Gutknecht & G. Kostova

  2. Department of Experimental Haematology, GSF-Forschungszentrum, D-81377, München, Germany

    K. Thalmeier

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  1. H. Porzig
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  2. R. Gutknecht
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  3. G. Kostova
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  4. K. Thalmeier
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Porzig, H., Gutknecht, R., Kostova, G. et al. G-protein-coupled receptors in normal human erythroid progenitor cells. Naunyn-Schmiedeberg's Arch Pharmacol 353, 11–20 (1995). https://doi.org/10.1007/BF00168910

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