Upregulation of the receptor-interacting protein 3 expression and involvement in neural tissue damage after spinal cord injury in mice

BMC Neuroscience

Eiji Itoi¹  Kenichiro Yahata¹  Satoshi Tateda¹  Hiroshi Ozawa¹  Haruo Kanno¹ 

[1] Department of Orthopaedic Surgery, Tohoku University School of Medicine, Seiryo-machi, Aoba-ku, Sendai 980-8574, Miyagi, Japan

关键词: Apoptosis;    Necrosis;    Cell death;    Necroptosis;    Spinal cord injury;    Receptor-interacting protein 3;   

Others  :  1230645
DOI  :  10.1186/s12868-015-0204-0

 received in 2014-11-22, accepted in 2015-10-01,  发布年份 2015

【 摘 要 】

Background

Necroptosis is a newly identified type of programmed cell death that differs from apoptosis. Recent studies have demonstrated that necroptosis is involved in multiple pathologies of various human diseases. Receptor-interacting protein 3 (RIP3) is known to be a critical regulator of necroptosis. This study investigated alterations in the RIP3 expression and the involvement in neural tissue damage after spinal cord injury (SCI) in mice.

Results

Immunohistochemical analysis demonstrated that the RIP3 expression was significantly increased in the lesion site after spinal cord hemisection. The increased expression of RIP3 started at 24 h, peaked at 3 days and lasted for at least 21 days after hemisection. The RIP3 expression was observed in neurons, astrocytes and oligodendrocytes. Western blot analysis also demonstrated the RIP3 protein expression significantly upregulated in the injured spinal cord. RIP3 staining using propidium iodide (PI)-labeled sections showed most of the PI-labeled cells were observed as RIP3-positive. Double staining of TUNEL and RIP3 demonstrated that TUNEL-positive cells exhibiting shrunken or fragmented nuclei, as generally observed in apoptotic cells, rarely expressed RIP3.

Conclusions

The present study first demonstrated that the expression of RIP3 is dramatically upregulated in various neural cells in the injured spinal cord and peaked at 3 days after injury. Additionally, most of the PI-labeled cells expressed RIP3 in response to neural tissue damage after SCI. The present study suggested that the upregulation of the RIP3 expression may play a role as a novel molecular mechanism in secondary neural tissue damage following SCI. However, further study is needed to clarify the specific molecular mechanism underlying the relationship between the RIP3 expression and cell death in the injured spinal cord.

【 授权许可】

   
2015 Kanno et al.

【 图 表 】

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【 参考文献 】

• [1]Clarke PG: Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol 1990, 181(3):195-213.
• [2]Puyal J, Ginet V, Clarke P: Multiple interacting cell death mechanisms in the mediation of excitotoxicity and ischemic brain damage: A challenge for neuroprotection. Prog Neurobiol 2013, 105:24-48.
• [3]Vandenabeele P, Galluzzi L, Vanden B, Kroemer G: Molecular mechanisms of necroptosis: an ordered cellular explosion. Nat Rev Mol Cell Biol 2010, 11:700-714.
• [4]Hotchkiss RS, Strasser A, McDunn JE, Swanson PE: Cell death. N Engl J Med 2009, 361(16):1570-1583.
• [5]Kerr JF, Wyllie AH, Currie AR: Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 1972, 26(4):239-257.
• [6]Hitomi J, Christofferson D, Ng A, Yao J, Degterev A, Xavier R, Yuan J: Identification of a molecular signaling network that regulates a cellular necrotic cell death pathway. Cell 2008, 135:1311-1323.
• [7]Degterev A, Hitomi J, Germscheid M, Ch’en I, Korkina O, Teng X, Abbott D, Cuny G, Yuan C, Wagner G, et al.: Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol 2008, 4(5):313-321.
• [8]Galluzzi L, Vitale I, Abrams J, Alnemri E, Baehrecke E, Blagosklonny M, Dawson T, Dawson V, El-Deiry W, Fulda S, et al.: Molecular definitions of cell death subroutines: recommendations of the Nomenclature Committee on Cell Death 2012. Cell Death Differ 2012, 19:107-120.
• [9]Han J, Zhong C, Zhang D: Programmed necrosis: backup to and competitor with apoptosis in the immune system. Nat Immunol 2011, 12:1143-1149.
• [10]Yang Y, Ma J, Chen Y, Wu M: Nucleocytoplasmic shuttling of receptor-interacting protein 3 (RIP3): identification of novel nuclear export and import signals in RIP3. J Biol Chem 2004, 279(37):38820-38829.
• [11]Galluzzi L, Vanden B, Vanlangenakker N, Buettner S, Eisenberg T, Vandenabeele P, Madeo F, Kroemer G: Programmed necrosis from molecules to health and disease. Int Rev Cell Mol Biol 2011, 289:1-35.
• [12]Zhang DW, Shao J, Lin J, Zhang N, Lu BJ, Lin SC, Dong MQ, Han J: RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science 2009, 325(5938):332-336.
• [13]Moriwaki K, Chan FKM: RIP3: a molecular switch for necrosis and inflammation. Genes Dev 2013, 27:1640-1649.
• [14]Roychowdhury S, McMullen M, Pisano S, Liu X, Nagy L: Absence of receptor interacting protein kinase 3 prevents ethanol-induced liver injury. Hepatology 2013, 57:1773-1783.
• [15]Kim L: Caspase blockade induces RIP3-mediated programmed necrosis in Toll-like receptor-activated microglia. Cell Death Dis 2013, 4:e716.
• [16]Lin J, Li H, Yang M, Ren J, Huang Z, Han F, Huang J, Ma J, Zhang D, Zhang Z, et al.: A role of RIP3-mediated macrophage necrosis in atherosclerosis development. Cell Rep 2013, 3(1):200-210.
• [17]Vieira M, Fernandes J, Carreto L, Anuncibay-Soto B, Santos M, Han J, Fernandez-Lopez A, Duarte CB, Carvalho AL, Santos AE: Ischemic insults induce necroptotic cell death in hippocampal neurons through the up-regulation of endogenous RIP3. Neurobiol Dis. 2014, 68:26-36.
• [18]Khan N, Lawlor KE, Murphy JM, Vince JE: More to life than death: molecular determinants of necroptotic and non-necroptotic RIP3 kinase signaling. Curr Opin Immunol 2014, 26:76-89.
• [19]Upton JW, Kaiser WJ, Mocarski ES: Virus inhibition of RIP3-dependent necrosis. Cell Host Microbe 2010, 7(4):302-313.
• [20]Smith CCT, Yellon DM: Necroptosis, necrostatins and tissue injury. J Cell Mol Med 2011, 15:1797-1806.
• [21]Günther C, Martini E, Wittkopf N, Amann K, Weigmann B, Neumann H, Waldner M, Hedrick S, Tenzer S, Neurath M, et al.: Caspase-8 regulates TNF-α-induced epithelial necroptosis and terminal ileitis. Nature 2011, 477:335-339.
• [22]Bonnet M, Preukschat D, Welz P, van Loo G, Ermolaeva M, Bloch W, Haase I, Pasparakis M: The adaptor protein FADD protects epidermal keratinocytes from necroptosis in vivo and prevents skin inflammation. Immunity 2011, 35:572-582.
• [23]Linkermann A, Bräsen J, Darding M, Jin M, Sanz A, Heller J, De Zen F, Weinlich R, Ortiz A, Walczak H, et al.: Two independent pathways of regulated necrosis mediate ischemia-reperfusion injury. Proc Natl Acad Sci USA 2013, 110:12024-12029.
• [24]Huang J, Shang L, Zhang M, Wang H, Chen D, Tong J, Huang H, Yan X, Zeng L, Xiong K: Differential neuronal expression of receptor interacting protein 3 in rat retina: involvement in ischemic stress response. BMC Neurosci 2013, 14:16. BioMed Central Full Text
• [25]Kanno H, Ozawa H, Sekiguchi A, Yamaya S, Itoi E: Induction of autophagy and autophagic cell death in damaged neural tissue after acute spinal cord injury in mice. Spine 2011, 36(22):E1427-E1434.
• [26]Kanno H, Ozawa H, Sekiguchi A, Itoi E: Spinal cord injury induces upregulation of Beclin 1 and promotes autophagic cell death. Neurobiol Dis 2009, 33(2):143-148.
• [27]Dong H, Fazzaro A, Xiang C, Korsmeyer SJ, Jacquin MF, McDonald JW: Enhanced oligodendrocyte survival after spinal cord injury in Bax-deficient mice and mice with delayed Wallerian degeneration. J Neurosci Off J Soc Neurosci 2003, 23(25):8682-8691.
• [28]Kalderon N, Fuks Z: Structural recovery in lesioned adult mammalian spinal cord by x-irradiation of the lesion site. Proc Natl Acad Sci USA 1996, 93(20):11179-11184.
• [29]You Z, Savitz S, Yang J, Degterev A, Yuan J, Cuny G, Moskowitz M, Whalen M: Necrostatin-1 reduces histopathology and improves functional outcome after controlled cortical impact in mice. J Cereb Blood Flow Metab 2008, 28:1564-1573.
• [30]Zhu X, Tao L, Tejima-Mandeville E, Qiu J, Park J, Garber K, Ericsson M, Lo E, Whalen M: Plasmalemma permeability and necrotic cell death phenotypes after intracerebral hemorrhage in mice. Stroke 2012, 43:524-531.
• [31]Whalen M, Dalkara T, You Z, Qiu J, Bermpohl D, Mehta N, Suter B, Bhide P, Lo E, Ericsson M, et al.: Acute plasmalemma permeability and protracted clearance of injured cells after controlled cortical impact in mice. J Cereb Blood Flow Metab 2008, 28:490-505.
• [32]Citron BA, Arnold PM, Sebastian C, Qin F, Malladi S, Ameenuddin S, Landis ME, Festoff BW: Rapid upregulation of caspase-3 in rat spinal cord after injury: mRNA, protein, and cellular localization correlates with apoptotic cell death. Exp Neurol 2000, 166(2):213-226.
• [33]Lu J, Ashwell KW, Waite P: Advances in secondary spinal cord injury: role of apoptosis. Spine 2000, 25(14):1859-1866.
• [34]Yong C, Arnold PM, Zoubine MN, Citron BA, Watanabe I, Berman NE, Festoff BW: Apoptosis in cellular compartments of rat spinal cord after severe contusion injury. J Neurotrauma 1998, 15(7):459-472.
• [35]Beattie MS, Farooqui AA, Bresnahan JC: Review of current evidence for apoptosis after spinal cord injury. J Neurotrauma 2000, 17(10):915-925.
• [36]Degterev A, Huang Z, Boyce M, Li Y, Jagtap P, Mizushima N, Cuny G, Mitchison T, Moskowitz M, Yuan J: Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nat Chem Biol 2005, 1:112-119.
• [37]Xu X, Chua K, Chua C, Liu C, Hamdy R, Chua B: Synergistic protective effects of humanin and necrostatin-1 on hypoxia and ischemia/reperfusion injury. Brain Res 2010, 1355:189-194.
• [38]Northington F, Chavez-Valdez R, Graham E, Razdan S, Gauda E, Martin L: Necrostatin decreases oxidative damage, inflammation, and injury after neonatal HI. J Cereb Blood Flow Metab 2011, 31:178-189.
• [39]Pirooznia SK, Dawson VL, Dawson TM: Motor neuron death in ALS: programmed by astrocytes? Neuron. 2014, 81:961-963.
• [40]Re LV, Yu A, Politi P, Ikiz H, Koolen N, Papadimitriou N, Mitsumoto K, Wichterle H, Przedborski : necroptosis drives motor neuron death in models of both sporadic and familial ALS. Neuron 2014, 81:1001-1008.
• [41]Rosenbaum D, Degterev A, David J, Rosenbaum P, Roth S, Grotta J, Cuny G, Yuan J, Savitz S: Necroptosis, a novel form of caspase-independent cell death, contributes to neuronal damage in a retinal ischemia-reperfusion injury model. J Neurosci Res 2010, 88:1569-1576.
• [42]Tristão V, Gonçalves P, Dalboni M, Batista M, Durão M, Martins J: Nec-1 protects against nonapoptotic cell death in cisplatin-induced kidney injury. Ren Fail 2012, 34:373-377.
• [43]MacVicar BA, Thompson RJ: Non-junction functions of pannexin-1 channels. Trends Neurosci 2010, 33(2):93-102.
• [44]Laird M, Wakade C, Alleyne J, Dhandapani K: Hemin-induced necroptosis involves glutathione depletion in mouse astrocytes. Free Radic Biol Med 2008, 45:1103-1114.
• [45]Simenc L-S: Staurosporine induces different cell death forms in cultured rat astrocytes. Radiol Oncol 2012, 46:312-320.
• [46]Kim S, Dayani L, Rosenberg P, Li J: RIP1 kinase mediates arachidonic acid-induced oxidative death of oligodendrocyte precursors. Int J Physiol Pathophysiol Pharmacol 2010, 2:137-147.