PMID- 24746856 OWN - NLM STAT- MEDLINE DCOM- 20150511 LR - 20161125 IS - 1095-953X (Electronic) IS - 0969-9961 (Linking) VI - 68 DP - 2014 Aug TI - Ischemic insults induce necroptotic cell death in hippocampal neurons through the up-regulation of endogenous RIP3. PG - 26-36 LID - S0969-9961(14)00084-9 [pii] LID - 10.1016/j.nbd.2014.04.002 [doi] AB - Global cerebral ischemia induces selective acute neuronal injury of the CA1 region of the hippocampus. The type of cell death that ensues may include different programmed cell death mechanisms namely apoptosis and necroptosis, a recently described type of programmed necrosis. We investigated whether necroptosis contributes to hippocampal neuronal death following oxygen-glucose deprivation (OGD), an in vitro model of global ischemia. We observed that OGD induced a death receptor (DR)-dependent component of necroptotic cell death in primary cultures of hippocampal neurons. Additionally, we found that this ischemic challenge upregulated the receptor-interacting protein kinase 3 (RIP3) mRNA and protein levels, with a concomitant increase of the RIP1 protein. Together, these two related proteins form the necrosome, the complex responsible for induction of necroptotic cell death. Interestingly, we found that caspase-8 mRNA, a known negative regulator of necroptosis, was transiently decreased following OGD. Importantly, we observed that the OGD-induced increase in the RIP3 protein was paralleled in an in vivo model of transient global cerebral ischemia, specifically in the CA1 area of the hippocampus. Moreover, we show that the induction of endogenous RIP3 protein levels influenced neuronal toxicity since we found that RIP3 knock-down (KD) abrogated the component of OGD-induced necrotic neuronal death while RIP3 overexpression exacerbated neuronal death following OGD. Overexpression of RIP1 also had deleterious effects following the OGD challenge. Taken together, our results highlight that cerebral ischemia activates transcriptional changes that lead to an increase in the endogenous RIP3 protein level which might contribute to the formation of the necrosome complex and to the subsequent component of necroptotic neuronal death that follows ischemic injury. CI - Copyright (c) 2014 Elsevier Inc. All rights reserved. FAU - Vieira, M AU - Vieira M AD - Center for Neuroscience and Cell Biology, University of Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Portugal. FAU - Fernandes, J AU - Fernandes J AD - Center for Neuroscience and Cell Biology, University of Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Portugal. FAU - Carreto, L AU - Carreto L AD - RNA Biology Laboratory, Department of Biology and CESAM, University of Aveiro, Portugal. FAU - Anuncibay-Soto, B AU - Anuncibay-Soto B AD - Area Biologia Celular, Instituto de Biomedicina, Universidad de Leon, Leon, Spain. FAU - Santos, M AU - Santos M AD - RNA Biology Laboratory, Department of Biology and CESAM, University of Aveiro, Portugal. FAU - Han, J AU - Han J AD - The Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, China. FAU - Fernandez-Lopez, A AU - Fernandez-Lopez A AD - Area Biologia Celular, Instituto de Biomedicina, Universidad de Leon, Leon, Spain. FAU - Duarte, C B AU - Duarte CB AD - Center for Neuroscience and Cell Biology, University of Coimbra, Portugal; Department of Life Sciences, Faculty of Science and Technology, University of Coimbra, Portugal. FAU - Carvalho, A L AU - Carvalho AL AD - Center for Neuroscience and Cell Biology, University of Coimbra, Portugal; Department of Life Sciences, Faculty of Science and Technology, University of Coimbra, Portugal. Electronic address: alc@cnc.uc.pt. FAU - Santos, A E AU - Santos AE AD - Center for Neuroscience and Cell Biology, University of Coimbra, Portugal; Faculty of Pharmacy, University of Coimbra, Portugal. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140416 PL - United States TA - Neurobiol Dis JT - Neurobiology of disease JID - 9500169 RN - 0 (Antibodies) RN - 0 (Imidazoles) RN - 0 (Indoles) RN - 0 (Neuroprotective Agents) RN - 0 (Tumor Necrosis Factor-alpha) RN - 0 (necrostatin-1) RN - 6LR8C1B66Q (Dizocilpine Maleate) RN - EC 1.1.1.27 (L-Lactate Dehydrogenase) RN - EC 2.7.11.1 (RIPK3 protein, human) RN - EC 2.7.11.1 (Receptor-Interacting Protein Serine-Threonine Kinases) RN - IY9XDZ35W2 (Glucose) SB - IM MH - Animals MH - Antibodies/pharmacology MH - Apoptosis/drug effects/*physiology MH - Brain Ischemia/metabolism/*pathology MH - Cells, Cultured MH - Disease Models, Animal MH - Dizocilpine Maleate/pharmacology MH - Embryo, Mammalian MH - Glucose/deficiency MH - Hippocampus/cytology/*pathology MH - Hypoxia/*metabolism/pathology MH - Imidazoles/pharmacology MH - Indoles/pharmacology MH - L-Lactate Dehydrogenase/metabolism MH - Neurons/drug effects/*pathology MH - Neuroprotective Agents/pharmacology MH - Rats MH - Rats, Sprague-Dawley MH - Receptor-Interacting Protein Serine-Threonine Kinases/genetics/*metabolism MH - Tumor Necrosis Factor-alpha/immunology/metabolism MH - Up-Regulation/drug effects/*physiology OTO - NOTNLM OT - Cerebral ischemia OT - Necroptosis OT - Necrostatin-1 OT - Oxygen-glucose deprivation OT - Receptor-interacting Protein Kinase 3 EDAT- 2014/04/22 06:00 MHDA- 2015/05/12 06:00 CRDT- 2014/04/22 06:00 PHST- 2013/12/05 00:00 [received] PHST- 2014/04/01 00:00 [revised] PHST- 2014/04/08 00:00 [accepted] PHST- 2014/04/22 06:00 [entrez] PHST- 2014/04/22 06:00 [pubmed] PHST- 2015/05/12 06:00 [medline] AID - S0969-9961(14)00084-9 [pii] AID - 10.1016/j.nbd.2014.04.002 [doi] PST - ppublish SO - Neurobiol Dis. 2014 Aug;68:26-36. doi: 10.1016/j.nbd.2014.04.002. Epub 2014 Apr 16.