Widespread Expression of Erythropoietin Receptor in Brain and Its Induction by Injury.
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 immuno-precipitated endogenous EPOR, Western blotting, immunostaining and flow cytometry. Among others, we applied genetic strategies including overexpression, Lentivirus-mediated conditional knockout ofEpoRand tagged proteins, both on cultured cells and tissue sections, as well as intracortical implantation ofEPOR-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.
Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Christoph
C
Ott
Synaptic Systems GmbH, Göttingen, Germany.
Henrik
H
Martens
Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.|Physiology Unit, Zoology Department, Faculty of Science, Menoufia University, Egypt.
Imam
I
Hassouna
Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Bárbara
B
Oliveira
Synaptic Systems GmbH, Göttingen, Germany.
Christian
C
Erck
Institute of Pharmacology, University Medicine Göttingen, Göttingen, Germany.
Maria-Patapia
MP
Zafeiriou
Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Ulla-Kaisa
UK
Peteri
Proteomics Group, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Dörte
D
Hesse
Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Simone
S
Gerhart
Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Bekir
B
Altas
Synaptic Systems GmbH, Göttingen, Germany.
Tekla
T
Kolbow
Synaptic Systems GmbH, Göttingen, Germany.
Herbert
H
Stadler
Molecular Neurobiology, Max Planck Institute of Experimental Medicine, Göttingen, Germany.
Hiroshi
H
Kawabe
Institute of Pharmacology, University Medicine Göttingen, Göttingen, Germany.
Wolfram-Hubertus
WH
Zimmermann
Neurogenetics, Max Planck Institute of Experimental Medicine, Göttingen, Germany.|DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Göttingen, Germany.
Klaus-Armin
KA
Nave
Department of Neuropathology, University Medicine Göttingen, Göttingen, Germany.
Walter
W
Schulz-Schaeffer
Proteomics Group, Max Planck Institute of Experimental Medicine, Göttingen, Germany.|DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Göttingen, Germany.
Olaf
O
Jahn
Clinical Neuroscience, Max Planck Institute of Experimental Medicine, Göttingen, Germany.|DFG Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), Göttingen, Germany.
Hannelore
H
Ehrenreich