mTOR/S6 kinase pathway contributes to astrocyte survival during ischemia.
Neurons are highly dependent on astrocyte survival during brain damage. To identify genes involved in astrocyte function during ischemia, we performed mRNA differential display in astrocytes after oxygen and glucose deprivation (OGD). We detected a robust down-regulation of S6 kinase 1 (S6K1) mRNA that was accompanied by a sharp decrease in protein levels and activity. OGD-induced apoptosis was increased by the combined deletion of S6K1 and S6K2 genes, as well as by treatment with rapamycin that inhibits S6K1 activity by acting on the upstream regulator mTOR (mammalian target of rapamycin). Astrocytes lacking S6K1 and S6K2 (S6K1;S6K2-/-) displayed a defect in BAD phosphorylation and in the expression of the anti-apoptotic factors Bcl-2 and Bcl-xL. Furthermore reactive oxygen species were increased while translation recovery was impaired in S6K-deficient astrocytes following OGD. Rescue of either S6K1 or S6K2 expression by adenoviral infection revealed that protective functions were specifically mediated by S6K1, because this isoform selectively promoted resistance to OGD and reduction of ROS levels. Finally, "in vivo" effects of S6K suppression were analyzed in the permanent middle cerebral artery occlusion model of ischemia, in which absence of S6K expression increased mortality and infarct volume. In summary, this article uncovers a protective role for astrocyte S6K1 against brain ischemia, indicating a functional pathway that senses nutrient and oxygen levels and may be beneficial for neuronal survival.
Pastor
María Dolores
MD
Department of Medical Sciences, Medical School, Universidad de Castilla La Mancha, 02006 Albacete, Spain.
García-Yébenes
Isaac
I
Department of Pharmacology, Medical School, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Fradejas
Noelia
N
Department of Medical Sciences, Medical School, Universidad de Castilla La Mancha, 02006 Albacete, Spain.
Pérez-Ortiz
José Manuel
JM
Department of Medical Sciences, Medical School, Universidad de Castilla La Mancha, 02006 Albacete, Spain.
Mora-Lee
Silvia
S
Department of Medical Sciences, Medical School, Universidad de Castilla La Mancha, 02006 Albacete, Spain.
Tranque
Pedro
P
Department of Medical Sciences, Medical School, Universidad de Castilla La Mancha, 02006 Albacete, Spain.
Moro
María Ángeles
MÁ
Department of Pharmacology, Medical School, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Pende
Mario
M
INSERM U845, Université Paris Descartes, 75015 Paris, France.
Calvo
Soledad
S
Department of Medical Sciences, Medical School, Universidad de Castilla La Mancha, 02006 Albacete, Spain.
D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002470:Cell Survival; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism; D007511:Ischemia / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D010100:Oxygen / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism*; D058570:TOR Serine-Threonine Kinases; D051020:bcl-X Protein / Q000378:metabolism
C494812:Bcl2l1 protein, mouse; D002352:Carrier Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051020:bcl-X Protein; D017853:Phosphotransferases (Alcohol Group Acceptor); C546843:mTOR protein, mouse; D019893:Ribosomal Protein S6 Kinases; D058570:TOR Serine-Threonine Kinases; D005947:Glucose; D010100:Oxygen