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Nitric oxide regulates AKT phosphorylation and nuclear translocation in cultured retinal cells

Telmo A Mejía-García1, Camila C Portugal, Thaísa G Encarnação

  • 1Program of Neurosciences and Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, RJ, Brazil.

Cellular Signalling
|August 21, 2013
PubMed

Insights

Nitric oxide (NO) activates AKT phosphorylation and nuclear translocation in retinal neurons, promoting cell survival. This pathway involves cyclic GMP/protein kinase G and phosphoinositide 3-kinase, crucial for central nervous system development.

Area of Science:

  • Neuroscience
  • Molecular Biology
  • Cell Signaling

Background:

  • Nitric oxide (NO) is known to inhibit retinal neuron apoptosis via cyclic GMP/protein kinase G (PKG) and kinase pathways like PI3k/AKT.
  • AKT and NO are crucial for central nervous system (CNS) development and plasticity, exhibiting survival-promoting properties.

Purpose of the Study:

  • To investigate the effects of exogenous NO (S-nitroso-N-acetylpenicillamin, SNAP) and endogenous NO (l-arginine) on AKT phosphorylation in cultured chick retinal neurons.
  • To elucidate the signaling pathways involved in NO-mediated AKT activation and its role in neuronal survival.

Main Methods:

  • Treatment of cultured chick retinal neurons with NO donors (SNAP) or precursors (l-arginine).
  • Assessment of AKT phosphorylation at serine-473 and threonine-308 using specific inhibitors (ODQ, LY83583, KT5823, LY294002, wortmannin, KU0063794) and activators (YC-1, 8-Bromo cyclic GMP, zaprinast).
  • Evaluation of NMDA receptor involvement and NO synthase inhibition. Analysis of AKT translocation to the nucleus and protection against hydrogen peroxide-induced cell death.

Main Results:

  • SNAP and l-arginine significantly enhanced AKT phosphorylation in a time- and concentration-dependent manner.
  • The observed effects were mediated by soluble guanylyl cyclase, PKG, phosphoinositide 3-kinase, and the mTORC2 complex.
  • Glutamate and NMDA promoted AKT phosphorylation, dependent on NO production. SNAP and l-arginine induced AKT nuclear translocation and protected retinal cells from oxidative stress.

Conclusions:

  • Nitric oxide, from both endogenous and exogenous sources, activates AKT signaling in retinal neurons.
  • NO-induced AKT activation involves the PKG and PI3k/mTORC2 pathways and leads to nuclear translocation of AKT.
  • This NO-mediated AKT activation pathway plays a significant role in neuronal survival, particularly during CNS development.
Keywords:
(5R,2S)-(1)5methyl-10,11-dihydro-5H-dibenzo [a,d] cyclohepten-5,10-imine hydrogen maleate1H-[1,2,4]oxadiazolo[4,3-aquinoxalin-1-one]3-(5′hydroxymethyl-2′-furyl)-1-1benzyl indazole5-(2-Propoxyphenyl)-1H-[1,2,3]triazolo[4,5-d]pyrimidin-7(4H)-oneAKTAMPACMFCREBChick retinaCyclic GMPEDTAERKKT5823KU0063794L-ArgL-NAMEMAPKMEMMK-801N-acetylcysteineN-methyl-d-aspartateN-nitro-l-arginine methyl esterNACNMDANMDA receptorNONOSODQPDPD98058PI3KPKGProtein kinase GS-nitroso-N-acetylpenicillaminSDS-PAGESNAPWorYC-1[1-oxo-9.12-epoxy-1H-diidolo[1,2,3-fg:3′,2′,1′kl]pyrrolo[3,4][1,6]benzodiazocine-10carboxylic acid methyl estera-amino-3-hydroxy-5-methylisoxazole-4-propionatecGMPcalcium and magnesium-free balanced salt solutioncyclic guanosine monophosphatecyclic nucleotide responsive element binding proteinethylene diamine tetraacetic acidextracellular signal-regulated kinasel-Argininel-argininemTORminimum essential mediummitogen-associated protein kinasenitric oxidenitric-oxide synthasephosphatidylinositol 3′ kinaseprotein kinase Bprotein kinase Grel-5-[2-[(2R,6S)-2,6-dimethyl-4-morpholinyl]-4-(4-morpholinyl)pyrido[2,3-d]pyrimidin-7-yl]-2-methoxybenzenemethanolsGCsodium-dodecyl sulfate polyacrylamide gel electrophoresissoluble guanylate cyclasewortmannin, (1S,6bR,9aS,11R,11bR) 11-(Acetyloxy)-1,6b,7,8,9a,10,11,11b-octahydro-1-(methoxymethyl)-9a,11b-dimethyl-3H-furo[4,3,2-de]indeno[4,5,-h]-2-h]-2-benzopyran-3,6,9-trionezaprinast

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