Abstract
In the central nervous system, astrocytes produce erythropoietin (Epo) and neurons express its receptor. To examine whether or not the brain Epo protects the in vitro cultured neurons from glutamate-induced cell death, we established rat astrocyte cell lines containing the plasmid for production of recombinant rat Epo. Epo partially purified from the culture medium showed a neuroprotective effect similar to that of rat Epo produced by Chinese hamster ovary (CHO) cells. Comparison was made in some other properties between Epo produced by these astrocyte cell lines and that by CHO cells.
Similar content being viewed by others
References
Benveniste H, Drejer J, Schousboe A and Diemer NH (1984) Elevation of the extracellular concentrations of glutamate and aspartate in rat hippocampus during transient cerebral ischemia monitored by intracerebral microdialysis. J Neurochem 43: 1369–1374.
Choi DW (1985) Glutamate neurotoxicity in cortical cell culture is calcium dependent. Neurosci Lett 58: 293–297.
Choi DW (1987) Ionic dependence of glutamate neurotoxicity. J Neurosci 7: 369–379.
Choi DW (1988) Glutamate neurotoxicity and diseases of the nervous system. Neuron 1: 623–634.
Choi DW and Rothman SM (1990) The role of glutamate neurotoxicity in hypoxic-ischemic neuronal death. Annu Rev Neurosci 13: 171–182.
Digicaylioglu M, Bichet S, Marti HH, Wenger RH, Rivas LA, Bauer C and Gassmann M (1995) Localization of specific erythropoietin binding sites in defined areas of the mouse brain. Proc Natl Acad Sci USA 92: 3717–3720.
Goto M, Akai K, Murakami A, Hashimoto C, Tsuda E, Ueda M, Kawanishi G, Takahashi N, Ishimoto A, Chiba H and Sasaki R (1988) Production of recombinant human erythropoietin in mammalian cells: Host-cell dependency of the biological activity of the cloned glycoprotein. Bio/Technology 6: 67–71.
Goto M, Murakami A, Akai K, Kawanishi G, Ueda M, Chiba H and Sasaki R (1989) Characterization and use of monoclonal antibodies directed against human erythropoietin that recognize different antigenic determinants. Blood 74: 1415–1423.
Jacobs K, Shoemaker C, Rudersdorf R, Neill SD, Kaufman RJ, Mufson A, Seehra J, Jones SS, Hewick R, Fritsch EF, Kawakita M, Shimizu T and Miyake T (1985) Isolation and characterization of genomic and cDNA clones of human erythropoietin. Nature 313: 806–810.
Jelkmann W (1992) Erythropoietin: structure, control of production, and function. Physiol Rev 72: 449–489.
Kambe T, Tada J, Chikuma M, Masuda S, Nagao M, Tsuchiya T, Ratcliffe PJ and Sasaki R (1998) Embryonal carcinoma P19 cells produce erythropoietin constitutively but express lactate dehydrogenase in an oxygen-dependent manner. Blood 91: 1185–1195.
Kirino T (1982) Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res 239: 57–69.
Krantz SB (1991) Erythropoietin. Blood 77: 419–434.
Lin FK, Suggs S, Lin CH, Browne JK, Smalling R, Egrie JC, Chen KK, Fox GM, Martin F, Stabinsky Z, Badrawi SM, Lai PH and Goldwasser E (1985) Cloning and expression of the human erythropoietin gene. Proc Natl Acad Sci USA 82: 7580–7584.
Lin CS, Lim SK, D'Agati V and Costantini F (1996) Differential effects of an erythropoietin receptor gene disruption on primitive and definitive erythropoiesis. Genes and Develop 10: 154–164.
Liu C, Shen K, Liu Z and Noguchi CT (1997) Regulated human erythropoietin receptor expression in mouse brain. J Biol Chem 272: 32395–32400.
Masuda S, Nagao M, Takahata K, Konishi Y, Gallyas Jr F, Tabira T and Sasaki R (1993) Functional erythropoietin receptor of the cells with neural characteristics: Comparison with receptor properties of erythroid cells. J Biol Chem 268: 11208–11216.
Masuda S, Okano M, Yamagishi K, Nagao M, Ueda M and Sasaki R (1994) A novel site of erythropoietin production: Oxygen-dependent production in cultured rat astrocytes. J Biol Chem 269: 19488–19493.
Masuda S, Chikuma M and Sasaki R (1997) Insulin-like growth factors and insulin stimulate erythropoietin production in primary cultured astrocytes. Brain Res 746: 63–70.
Marti HH, Wenger RH, Rivas LA, Straumann U, Digicaylioglu M, Henn V, Yonekawa Y, Bauer C and Gassmann M (1996) Erythropoietin gene expression in human, monkey and murine brain. Eur J Neurosci 8: 666–676.
Morishita E, Narita H, Nishida M, Kawashima N, Yamagishi K, Masuda S, Nagao M, Hatta H and Sasaki R (1996) Anti-erythropoietin receptor monoclonal antibody: Epitope mapping, quantification of the soluble receptor, and detection of the solubilized transmembrane receptor and the receptor-expressing cells. Blood 88: 465–471.
Morishita E, Masuda S, Nagao M, Yasuda Y and Sasaki R (1997) Erythropoietin receptor is expressed in rat hippocampal and cerebral cortical neurons, and erythropoietin prevents in vitro glutamate-induced neuronal death. Neuroscience 76: 105–116.
Nagao M, Masuda S, Abe S, Ueda M and Sasaki R (1992) Production and ligand-binding characteristics of the soluble form of murine erythropoietin receptor. Biochem Biophys Res Commun 188: 888–897.
Nagao M, Suga H, Okano M, Masuda S, Narita H, Ikura K and Sasaki R (1992) Nucleotide sequence of rat erythropoietin. Biochim Biophys Acta 1171: 99–102.
Ohashi H, Maruyama K, Liu YC and Yoshimura A. (1994) Ligand-induced activation of chimeric receptors between the erythropoietin receptor and receptor tyrosine kinases. Proc Natl Acad Sci USA 91: 158–162.
Pulsinelli WA, Brierly JB and Plum F (1982) Temporal profile of neuronal damage in a model of transient forebrain ischemia. Ann Neurol 11: 491–498.
Sakanaka M, Wen TC, Matsuda S, Masuda S, Morishita E, Nagao M and Sasaki R (1998) In vivo evidence that erythropoietin protects neurons from ischemic damage. Proc Natl Acad Sci USA 95: 4635–4640.
Siesjo BK (1988) Historical overview: Calcium, ischemia and death of brain cells. Ann NY Acad Sci 522: 638–661.
Tsuda E, Kawanishi G, Ueda M, Masuda S and Sasaki R (1990) The role of carbohydrate in recombinant human erythropoietin. Eur J Biochem 188: 405–411.
Wu H, Liu X, Jaenisch R and Lodish HF (1995) Generation of committed erythroid BFU-E and CFU-E progenitors does not require erythropoietin or the erythropoietin receptor. Cell 83: 59–67.
Yamaguchi K, Akai K, Kawanishi G, Ueda M, Masuda S and Sasaki R (1991) Effects of site-directed removal of N-glycosylation sites in human erythropoietin on its production and biological properties. J Biol Chem 266: 20434–20439.
Youssoufian H, Longmore G, Neumann D, Yoshimura A and Lodish HF (1993) Structure, function, and activation of the erythropoietin receptor. Blood 81: 2223–2236.
Rights and permissions
About this article
Cite this article
Masuda, S., Kada, E., Nagao, M. et al. In vitro neuroprotective action of recombinant rat erythropoietin produced by astrocyte cell lines and comparative studies with erythropoietin produced by Chinese hamster ovary cells. Cytotechnology 31, 179–184 (1999). https://doi.org/10.1023/A:1008028423510
Issue Date:
DOI: https://doi.org/10.1023/A:1008028423510