title: The nitric oxide-cGKII system relays death and survival signals during embryonic retinal development via AKT-induced CREB1 activation.
abstract: During early neurogenesis, retinal neuronal cells display a conserved differentiation program in vertebrates. Previous studies established that nitric oxide (NO) and cGMP accumulation regulate essential events in retinal physiology. Here we used pharmacological and genetic loss-of-function to investigate the effects of NO and its downstream signaling pathway in the survival of developing avian retinal neurons in vitro and in vivo. Six-day-old (E6) chick retinal cells displayed increased calcium influx and produced higher amounts of NO when compared with E8 cells. L-arginine (substrate for NO biosynthesis) and S-nitroso-N-acetyl-D,L-penicillamine (SNAP; a nitrosothiol NO donor) promoted extensive cell death in E6 retinas, whereas in E8 both substances decreased apoptosis. The effect of NO at both periods was mediated by soluble guanylyl cyclase (sGC) and cGMP-dependent kinase (cGK) activation. In addition, shRNA-mediated cGKII knockdown prevented NO-induced cell death (E6) and cell survival (E8). This, NO-induced cell death or cell survival was not correlated with an early inhibition of retinal cell proliferation. E6 cells also responded differentially from E8 neurons regarding cyclic AMP-responsive element-binding protein (CREB) activation in the retina in vivo. NO strongly decreased nuclear phospho-CREB staining in E6 but it robustly enhanced CREB phosphorylation in the nuclei of E8 neurons, an effect that was completely abrogated by cGKII shRNAs at both embryonic stages. The ability of NO in regulating CREB differentially during retinal development relied on the capacity of cGKII in decreasing (E6) or increasing (E8) nuclear AKT (V-Akt murine thymoma viral oncogene) activation. Accordingly, inhibiting AKT prevented both cGKII shRNA-mediated CREB upregulation in E6 and SNAP-induced CREB activation in E8. Furthermore, shRNA-mediated in vivo cGKII or in vitro CREB1 knockdown confirmed that NO/cGKII dualistically regulated the downstream CREB1 pathway and caspase activation in the chick retina to modulate neuronal viability. These data demonstrate that NO-mediated cGKII signaling may function to control the viability of neuronal cells during early retinal development via AKT/CREB1 activity.
authors: Program of Neurosciences, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil.
authors: R
authors: R
authors: Socodato
authors: Program of Neurosciences, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil.
authors: R
authors: R
authors: Brito
authors: Program of Neurosciences, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil.
authors: C C
authors: CC
authors: Portugal
authors: Program of Neurosciences, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil.
authors: N A
authors: NA
authors: de Oliveira
authors: 1] Program of Neurosciences, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil [2] Department of Neurobiology, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil.
authors: K C
authors: KC
authors: Calaza
authors: 1] Program of Neurosciences, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil [2] Department of Neurobiology, Institute of Biology, Fluminense Federal University, Rio de Janeiro, Brazil.
authors: R
authors: R
authors: Paes-de-Carvalho
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001120:Arginine / Q000378:metabolism; D002470:Cell Survival / Q000235:genetics; D002642:Chick Embryo; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism*; D062967:Cyclic GMP-Dependent Protein Kinase Type II / Q000235:genetics / Q000378:metabolism*; D047108:Embryonic Development; D055495:Neurogenesis / Q000235:genetics; D009569:Nitric Oxide / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D012160:Retina / Q000254:growth & development* / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics
chemical_list: D017362:Cyclic AMP Response Element-Binding Protein; D009569:Nitric Oxide; D001120:Arginine; D051057:Proto-Oncogene Proteins c-akt; D062967:Cyclic GMP-Dependent Protein Kinase Type II