caa a22 4500 606177248 CHVBK 20210128100751.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00018-015-1927-x doi (NATIONALLICENCE)springer-10.1007/s00018-015-1927-x Apaf1-deficient cortical neurons exhibit defects in axonal outgrowth [Elektronische Daten] [Daniela De Zio, Francesca Molinari, Salvatore Rizza, Lucia Gatta, Maria Ciotti, Anna Salvatore, Søs Mathiassen, Andrzej Cwetsch, Giuseppe Filomeni, Giuseppe Rosano, Elisabetta Ferraro] The establishment of neuronal polarity and axonal outgrowth are key processes affecting neuronal migration and synapse formation, their impairment likely leading to cognitive deficits. Here we have found that the apoptotic protease activating factor 1 (Apaf1), apart from its canonical role in apoptosis, plays an additional function in cortical neurons, where its deficiency specifically impairs axonal growth. Given the central role played by centrosomes and microtubules in the polarized extension of the axon, our data suggest that Apaf1-deletion affects axonal outgrowth through an impairment of centrosome organization. In line with this, centrosomal protein expression, as well as their centrosomal localization proved to be altered upon Apaf1-deletion. Strikingly, we also found that Apaf1-loss affects trans-Golgi components and leads to a robust activation of AMP-dependent protein kinase (AMPK), this confirming the stressful conditions induced by Apaf1-deficiency. Since AMPK hyper-phosphorylation is known to impair a proper axon elongation, our finding contributes to explain the effect of Apaf1-deficiency on axogenesis. We also discovered that the signaling pathways mediating axonal growth and involving glycogen synthase kinase-3β, liver kinase B1, and collapsing-response mediator protein-2 are altered in Apaf1-KO neurons. Overall, our results reveal a novel non-apoptotic role for Apaf1 in axonal outgrowth, suggesting that the neuronal phenotype due to Apaf1-deletion could not only be fully ascribed to apoptosis inhibition, but might also be the result of defects in axogenesis. The discovery of new molecules involved in axonal elongation has a clinical relevance since it might help to explain neurological abnormalities occurring during early brain development. Springer Basel, 2015 Centrosome nationallicence Golgi nationallicence Rab GTPases nationallicence NF1 nationallicence Mitochondria nationallicence Neuro-rehabilitation nationallicence ACC : Acetyl-CoA carboxylase nationallicence AMPK : AMP-dependent protein kinase nationallicence Apaf1 : Apoptotic protease activating factor 1 nationallicence CRMP2 : Collapsing-response mediator protein-2 nationallicence DIV : Day in vitro nationallicence Diva : Death inducer binding to vBcl2 and Apaf1 nationallicence ETNA : Embryonic telencephalic naïve Apaf1 nationallicence Gap43 : Growth associated protein 43 nationallicence GDI : GDP dissociation inhibitor nationallicence GM130 : cis-Golgi marker nationallicence GSK3β : Glycogen synthase kinase-3β nationallicence HCA66 : Hepatocellular carcinoma-associated antigen 66 nationallicence I-MEFs : Immortalized mouse embryonic fibroblasts nationallicence LKB1 : Liver kinase B1 nationallicence MAP2 : Microtubule-associated protein 2 nationallicence MAPs : Microtubule-associated proteins nationallicence MARK : Microtubule affinity-regulating kinase nationallicence NEDD1 : Neural precursor cell expressed developmentally down-regulated protein 1 nationallicence NF1 : Neurofibromatosis type I nationallicence PCN : Primary cortical neurons nationallicence PSD95 : Postsynaptic density protein 95 nationallicence Rab8 : Ras-related in brain 8 nationallicence Rab10 : Ras-related in brain 10 nationallicence SMI312 : Pan-axonal neurofilament marker nationallicence Tau : Tau protein nationallicence Tom20 : Translocase of outer membrane 20 nationallicence Tubb3 : Tubulin, beta 3 class III nationallicence De Zio Daniela Department of Biology, "Tor Vergata” University of Rome, Via della Ricerca Scientifica, 00133, Rome, Italy aut Molinari Francesca Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut Rizza Salvatore Department of Biology, "Tor Vergata” University of Rome, Via della Ricerca Scientifica, 00133, Rome, Italy aut Gatta Lucia Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut Ciotti Maria Institute of Cell Biology and Neurobiology (IBCN), National Research Council (CNR), Rome, Italy aut Salvatore Anna Institute of Neurobiology and Molecular Medicine, National Research Council (CNR), Rome, Italy aut Mathiassen Søs Cell Stress and Survival Unit, Danish Cancer Society Research Center, Strandboulevarden 49, 2100, Copenhagen, Denmark aut Cwetsch Andrzej Department of Neuroscience and Brain Technologies, Italian Institute of Technology (IIT), via Morego 30, 16163, Genoa, Italy aut Filomeni Giuseppe Department of Biology, "Tor Vergata” University of Rome, Via della Ricerca Scientifica, 00133, Rome, Italy aut Rosano Giuseppe Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut Ferraro Elisabetta Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut Cellular and Molecular Life Sciences Springer Basel 72/21(2015-11-01), 4173-4191 1420-682X 72:21<4173 2015 72 18 https://doi.org/10.1007/s00018-015-1927-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1927-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- De Zio Daniela Department of Biology, "Tor Vergata” University of Rome, Via della Ricerca Scientifica, 00133, Rome, Italy aut NATIONALLICENCE 700 1- Molinari Francesca Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut NATIONALLICENCE 700 1- Rizza Salvatore Department of Biology, "Tor Vergata” University of Rome, Via della Ricerca Scientifica, 00133, Rome, Italy aut NATIONALLICENCE 700 1- Gatta Lucia Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut NATIONALLICENCE 700 1- Ciotti Maria Institute of Cell Biology and Neurobiology (IBCN), National Research Council (CNR), Rome, Italy aut NATIONALLICENCE 700 1- Salvatore Anna Institute of Neurobiology and Molecular Medicine, National Research Council (CNR), Rome, Italy aut NATIONALLICENCE 700 1- Mathiassen Søs Cell Stress and Survival Unit, Danish Cancer Society Research Center, Strandboulevarden 49, 2100, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Cwetsch Andrzej Department of Neuroscience and Brain Technologies, Italian Institute of Technology (IIT), via Morego 30, 16163, Genoa, Italy aut NATIONALLICENCE 700 1- Filomeni Giuseppe Department of Biology, "Tor Vergata” University of Rome, Via della Ricerca Scientifica, 00133, Rome, Italy aut NATIONALLICENCE 700 1- Rosano Giuseppe Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut NATIONALLICENCE 700 1- Ferraro Elisabetta Laboratory of Skeletal Muscle Development and Metabolism, IRCCS San Raffaele Pisana, Via di Val Cannuta 247, 00166, Rome, Italy aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/21(2015-11-01), 4173-4191 1420-682X 72:21<4173 2015 72 18