Modulation of calcium channels in human erythroblasts by erythropoietin

Joseph Y. Cheung, Xue Qian Zhang, Krister Bokvist, Douglas L. Tillotson, Barbara A. Miller

Research output: Contribution to journalArticle

44 Scopus citations

Abstract

Erythropoietin (Epo) induces a dose-dependent increase in intracellular free Ca2+ ([Ca2+](i)) in human erythroblasts, which is dependent on extracellular Ca2+ and blocked by high doses of nifedipine or Ni2+. In addition, pretreatment of human erythroblasts with mouse antihuman erythropoietin receptor antibody but not mouse immunopure IgG blocked the Epo-induced [Ca2+], increase, indicating the specificity of the Ca2+ response to Epo stimulation. In this study, the erythropoietin-regulated calcium channel was identified by single channel recordings. Use of conventional whole cell patch-clamp failed to detect Epo-induced whole cell Ca2+ current. To minimize washout of cytosolic constituents, we next used nystatin perforated patch, but did not find any Epo-induced whole cell Ca2+ current. Using Ba2+ (30 mmol/L) as charge carrier in cell-attached patches, we detected single channels with unitary conductance of 3.2 pS, reversal potential of +72 mV, and whose unitary current (at +10 mV) increased monotonically with increasing Ba2+ concentrations. Channel open probability did not appreciably change over the voltage range (-50 to +30 mV) tested. Epo (2 U/mL) increased both mean open time (from 4.27 ± 0.75 to 11.15 ± 1.80 ms) and open probability (from 0.26 ± 0.06 to 2.56 ± 0.59%) of this Ba2+permeable channel. Our data strongly support the conclusion that the Epo-induced [Ca2+](i) increase in human erythroblasts is mediated via Ca2+ entry through a voltage-independent Ca2+ channel.

Original languageEnglish (US)
Pages (from-to)92-100
Number of pages9
JournalBlood
Volume89
Issue number1
DOIs
StatePublished - Jan 1 1997

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Fingerprint Dive into the research topics of 'Modulation of calcium channels in human erythroblasts by erythropoietin'. Together they form a unique fingerprint.

  • Cite this