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Molecular Medicine

, Volume 21, Issue 1, pp 803–815 | Cite as

Widespread Expression of Erythropoietin Receptor in Brain and Its Induction by Injury

  • Christoph Ott
  • Henrik Martens
  • Imam Hassouna
  • Barbara Oliveira
  • Christian Erck
  • Maria-Patapia Zafeiriou
  • Ulla-Kaisa Peteri
  • Dörte Hesse
  • Simone Gerhart
  • Bekir Altas
  • Tekla Kolbow
  • Herbert Stadler
  • Hiroshi Kawabe
  • Wolfram-Hubertus Zimmermann
  • Klaus-Armin Nave
  • Walter Schulz-Schaeffer
  • Olaf Jahn
  • Hannelore Ehrenreich
Research Article

Abstract

Erythropoietin (EPO) exerts potent neuroprotective, neuroregenerative and procognitive functions. However, unequivocal demonstration of erythropoietin receptor (EPOR) expression in brain cells has remained difficult since previously available anti-EPOR antibodies (EPOR-AB) were unspecific. We report here a new, highly specific, polyclonal rabbit EPOR-AB directed against different epitopes in the cytoplasmic tail of human and murine EPOR and its characterization by mass spectrometric analysis of immunoprecipitated endogenous EPOR, Western blotting, immunostaining and flow cytometry. Among others, we applied genetic strategies including overexpression, Lentivirus-mediated conditional knockout of EpoR and tagged proteins, both on cultured cells and tissue sections, as well as intracortical implantation of EPOR-transduced cells to verify specificity. We show examples of EPOR expression in neurons, oligodendroglia, astrocytes and microglia. Employing this new EPOR-AB with double-labeling strategies, we demonstrate membrane expression of EPOR as well as its localization in intracellular compartments such as the Golgi apparatus. Moreover, we show injury-induced expression of EPOR. In mice, a stereotactically applied stab wound to the motor cortex leads to distinct EpoR expression by reactive GFAP-expressing cells in the lesion vicinity. In a patient suffering from epilepsy, neurons and oligodendrocytes of the hippocampus strongly express EPOR. To conclude, this new analytical tool will allow neuroscientists to pinpoint EPOR expression in cells of the nervous system and to better understand its role in healthy conditions, including brain development, as well as under pathological circumstances, such as upregulation upon distress and injury.

Notes

Acknowledgments

This work was supported by ZIM-BWMi, the Max Planck Society, the Deutsche Forschungsgemeinschaft (Center for Nanoscale Microscopy and Molecular Physiology of the Brain, SFB-TRR43), EXTRABRAIN EU-FP7 and the Niedersachsen-Research Network on Neuroinfectiology (N-RENNT). The EPO-dependent megakaryoblastic leukemia UT-7 cell line was a kind gift from Drorit Neumann of Tel Aviv University in Israel. The pMOWS vector, encoding N-terminally HA-tagged full-length human EPOR plus puromycinresistant cassette, was a kind gift from Ursula Klingmüller, DKFZ, Heidelberg, Germany. The rat anti-NG2 was a kind gift from Jacqueline Trotter, University of Mainz, Germany. We thank Marina Uecker and Thomas Liepold for their expert technical help.

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© The Author(s) 2015

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Authors and Affiliations

  • Christoph Ott
    • 1
  • Henrik Martens
    • 2
  • Imam Hassouna
    • 1
    • 3
  • Barbara Oliveira
    • 1
  • Christian Erck
    • 2
  • Maria-Patapia Zafeiriou
    • 4
  • Ulla-Kaisa Peteri
    • 1
  • Dörte Hesse
    • 5
  • Simone Gerhart
    • 1
  • Bekir Altas
    • 6
  • Tekla Kolbow
    • 2
  • Herbert Stadler
    • 2
  • Hiroshi Kawabe
    • 6
  • Wolfram-Hubertus Zimmermann
    • 4
  • Klaus-Armin Nave
    • 7
    • 8
  • Walter Schulz-Schaeffer
    • 9
  • Olaf Jahn
    • 5
    • 8
  • Hannelore Ehrenreich
    • 1
    • 8
  1. 1.Clinical NeuroscienceMax Planck Institute of Experimental MedicineGöttingenGermany
  2. 2.Synaptic Systems GmbHGöttingenGermany
  3. 3.Physiology Unit, Zoology Department, Faculty of ScienceMenoufia UniversityMenoufiaEgypt
  4. 4.Institute of PharmacologyUniversity Medicine GöttingenGöttingenGermany
  5. 5.Proteomics GroupMax Planck Institute of Experimental MedicineGöttingenGermany
  6. 6.Molecular NeurobiologyMax Planck Institute of Experimental MedicineGöttingenGermany
  7. 7.NeurogeneticsMax Planck Institute of Experimental MedicineGöttingenGermany
  8. 8.DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB)GöttingenGermany
  9. 9.Department of NeuropathologyUniversity Medicine GöttingenGöttingenGermany

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