BDNF Overexpression Enhances the Preconditioning Effect of Brief Episodes of Hypoxia,Promoting Survival of GABAergic Neurons

Hypoxia causes depression of synaptic plasticity,hyperexcitation of neuronal networks,and the death of specific populations of neurons.However,brief episodes of hypoxia can promote the adaptation of cells.Hypoxic preconditioning is well manifested in glutamatergic neurons,while this adaptive mechanism is virtually suppressed in GABAergic neurons.Here,we show that brain-derived neurotrophic factor(BDNF) overexpression in neurons enhances the preconditioning effect of brief episodes of hypoxia.The amplitudes of the NMDAR-and AMPARmediated Ca2+ responses of glutamatergic and GABAergic neurons gradually decreased after repetitive brief hypoxia/reoxygenation cycles in cell cultures transduced with the(AAV)-Syn-BDNF-EGFP virus construct.In contrast,the amplitudes of the responses of GABAergic neurons increased in non-transduced cultures after preconditioning.The decrease of the amplitudes in GABAergic neurons indicated the activation of mechanisms of hypoxic preconditioning.Preconditioning suppressed apoptotic or necrotic cell death.This effect was most pronounced in cultures with BDNF overe xpression.Knockdown of BDNF abolished the effect of preconditioning and promoted the death of GABAergic neurons.Moreover,the expression of the anti-apoptotic genes Stat3,Socs3,and Bcl-x1 substantially increased 24 h after hypoxic episodes in the transduced cultures compared to controls.The expression of genes encoding the pro-inflammatory cytokines IL-10 and IL-6 also increased.In turn,the expression of pro-apoptotic(Bax,Casp-3,and Fas) and proinflammatory(IL-1β and TNFα) genes decreased after hypoxic episodes in cultures with BDNF overexpression.Inhibition of vesicular BDNF release abolished its protective action targeting inhibition of the oxygen-glucose deprivation(OGD)-induced [Ca2+]i increase in GABAergic and glutamatergic neurons,thus promoting their death.Bafilomycin A1,Brefeldin A,and tetanus toxin suppressed vesicular release(including BDNF) and shifted the gene expression profile towards excitotoxicity,inflammation,and apoptosis.These inhibitors of vesicular release abolished the protective effects of hypoxic preconditioning in glutamatergic neurons24 h after hypoxia/reoxygenation cycles.This finding indicates a significant contribution of vesicular BDNF release to the development of the mechanisms of hypoxic preconditioning.Thus,our results demonstrate that BDNF plays a pivotal role in the activation and enhancement of the preconditioning effect of brief episodes of hypoxia and promotes tolerance of the most vulnerable populations of GABAergic neurons to hypoxia/ischemia.