Comparison of neuroprotective effects of erythropoietin (EPO) and carbamylerythropoietin (CEPO) against ischemia-like oxygen-glucose deprivation (OGD) and NMDA excitotoxicity in mouse hippocampal slice cultures.
In addition to its well-known hematopoietic effects, erythropoietin (EPO) also has neuroprotective properties. However, hematopoietic side effects are unwanted for neuroprotection, underlining the need for EPO-like compounds with selective neuroprotective actions. One such compound, devoid of hematopoietic bioactivity, is the chemically modified, EPO-derivative carbamylerythropoietin (CEPO). For comparison of the neuroprotective effects of CEPO and EPO, we subjected organotypic hippocampal slice cultures to oxygen-glucose deprivation (OGD) or N-methyl-d-aspartate (NMDA) excitotoxicity. Hippocampal slice cultures were pretreated for 24 h with 100 IU/ml EPO (=26 nM) or 26 nM CEPO before OGD or NMDA lesioning. Exposure to EPO and CEPO continued during OGD and for the next 24 h until histology, as well as during the 24 h exposure to NMDA. Neuronal cell death was quantified by cellular uptake of propidium iodide (PI), recorded before the start of OGD and NMDA exposure and 24 h after. In cultures exposed to OGD or NMDA, CEPO reduced PI uptake by 49+/-3 or 35+/-8%, respectively, compared to lesion-only controls. EPO reduced PI uptake by 33+/-5 and 15+/-8%, respectively, in the OGD and NMDA exposed cultures. To elucidate a possible mechanism involved in EPO and CEPO neuroprotection against OGD, the integrity of alpha-II-spectrin cytoskeletal protein was studied. Both EPO and CEPO significantly reduced formation of spectrin cleavage products in the OGD model. We conclude that CEPO is at least as efficient neuroprotectant as EPO when excitotoxicity is modeled in mouse hippocampal slice cultures.
Montero
Maria
M
Department of Anatomy and Neurobiology, Institute of Medical Biology, University of Southern Denmark, Winsløwparken 21 st, DK-5000 Odense C, Denmark. mmontero@health.sdu.dk
Poulsen
Frantz Rom
FR
Noraberg
Jens
J
Kirkeby
Agnete
A
van Beek
Johan
J
Leist
Marcel
M
Zimmer
Jens
J
D000818:Animals; D000831:Animals, Newborn; D002545:Brain Ischemia / Q000378:metabolism / Q000473:pathology; D002154:Calpain / Q000037:antagonists & inhibitors; D004396:Coloring Agents; D004921:Erythropoietin / Q000031:analogs & derivatives* / Q000494:pharmacology*; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology*; D005947:Glucose / Q000172:deficiency*; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002534:Hypoxia, Brain / Q000378:metabolism* / Q000473:pathology; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D016202:N-Methylaspartate / Q000494:pharmacology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011419:Propidium / Q000493:pharmacokinetics; D014048:Tolonium Chloride
D004396:Coloring Agents; D018690:Excitatory Amino Acid Agonists; D008869:Microtubule-Associated Proteins; D018696:Neuroprotective Agents; C488171:carbamylated erythropoietin; D004921:Erythropoietin; D014048:Tolonium Chloride; D011419:Propidium; D016202:N-Methylaspartate; D002154:Calpain; D005947:Glucose