title: Essential roles of neutral ceramidase and sphingosine in mitochondrial dysfunction due to traumatic brain injury.
abstract: In addition to immediate brain damage, traumatic brain injury (TBI) initiates a cascade of pathophysiological events producing secondary injury. The biochemical and cellular mechanisms that comprise secondary injury are not entirely understood. Herein, we report a substantial deregulation of cerebral sphingolipid metabolism in a mouse model of TBI. Sphingolipid profile analysis demonstrated increases in sphingomyelin species and sphingosine concurrently with up-regulation of intermediates of de novo sphingolipid biosynthesis in the brain. Investigation of intracellular sites of sphingosine accumulation revealed an elevation of sphingosine in mitochondria due to the activation of neutral ceramidase (NCDase) and the reduced activity of sphingosine kinase 2 (SphK2). The lack of change in gene expression suggested that post-translational mechanisms are responsible for the shift in the activities of both enzymes. Immunoprecipitation studies revealed that SphK2 is complexed with NCDase and cytochrome oxidase (COX) subunit 1 in mitochondria and that brain injury hindered SphK2 association with the complex. Functional studies showed that sphingosine accumulation resulted in a decreased activity of COX, a rate-limiting enzyme of the mitochondrial electron transport chain. Knocking down NCDase reduced sphingosine accumulation in mitochondria and preserved COX activity after the brain injury. Also, NCDase knockdown improved brain function recovery and lessened brain contusion volume after trauma. These studies highlight a novel mechanism of secondary TBI involving a disturbance of sphingolipid-metabolizing enzymes in mitochondria and suggest a critical role for mitochondrial sphingosine in promoting brain injury after trauma.
authors.affiliation: From the Ralph H. Johnson Veterans Affairs Medical Center, Charleston, South Carolina 29401.
authors.forename: Sergei A
authors.initials: SA
authors.lastname: Novgorodov
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Riley
authors.forename: Jin
authors.initials: J
authors.lastname: Yu
authors.forename: Keith T
authors.initials: KT
authors.lastname: Borg
authors.forename: Yusuf A
authors.initials: YA
authors.lastname: Hannun
authors.forename: Richard L
authors.initials: RL
authors.lastname: Proia
authors.forename: Mark S
authors.initials: MS
authors.lastname: Kindy
authors.forename: Tatyana I
authors.initials: TI
authors.lastname: Gudz
mesh_terms: D055574:Alkaline Ceramidase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D001930:Brain Injuries / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003576:Electron Transport Complex IV / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism; D013110:Sphingosine / Q000235:genetics / Q000378:metabolism*; D000099157:Sphingosine Kinase
chemical_list: D055574:Alkaline Ceramidase; D003576:Electron Transport Complex IV; D009419:Nerve Tissue Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); D013110:Sphingosine; D000099157:Sphingosine Kinase
keywords: Brain; Cell Death; Mitochondria; Respiratory Chain; Sphingolipid