Induction of serpinb1a by PACAP or NGF is required for PC12 cells survival after serum withdrawal

Authors

  • Tommy Seaborn,

    1. Neurotrophic Factor and Neuronal Differentiation Team, Inserm U982, DC2N, France
    2. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    3. Department of Pediatrics, Hôpital St-François d'Assise, Centre de Recherche du Centre Hospitalier Universitaire de Québec (CRCHUQ), Laval University, Québec, Canada
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    • These authors contributed equally to this work.
  • Aurélia Ravni,

    1. Neurotrophic Factor and Neuronal Differentiation Team, Inserm U982, DC2N, France
    2. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    3. PRIMACEN, IRIB, University of Rouen, Mont-Saint-Aignan, France
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    • These authors contributed equally to this work.
  • Ruby Au,

    1. Neurotrophic Factor and Neuronal Differentiation Team, Inserm U982, DC2N, France
    2. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    3. Department of Zoology, University of Hong Kong, Hong Kong, China
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  • Bill K. C. Chow,

    1. Department of Zoology, University of Hong Kong, Hong Kong, China
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  • Alain Fournier,

    1. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    2. Institut National de la Recherche Scientifique-Institut Armand Frappier, Université du Québec, Pointe-Claire, Canada
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  • Olivier Wurtz,

    1. Neurotrophic Factor and Neuronal Differentiation Team, Inserm U982, DC2N, France
    2. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    3. PRIMACEN, IRIB, University of Rouen, Mont-Saint-Aignan, France
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  • Hubert Vaudry,

    1. Neurotrophic Factor and Neuronal Differentiation Team, Inserm U982, DC2N, France
    2. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    3. PRIMACEN, IRIB, University of Rouen, Mont-Saint-Aignan, France
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  • Lee E. Eiden,

    1. Laboratory of Cellular and Molecular Regulation, National Institute of Mental Health, Bethesda, Maryland, USA
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  • David Vaudry

    Corresponding author
    1. Neurotrophic Factor and Neuronal Differentiation Team, Inserm U982, DC2N, France
    2. International Associated Laboratory Samuel de Champlain, Mont-Saint-Aignan, France
    3. PRIMACEN, IRIB, University of Rouen, Mont-Saint-Aignan, France
    • Address for correspondence and reprint requests to David Vaudry, Neurotrophic Factor and Neuronal Differentiation team, Inserm U982, University of Rouen, 76821 Mont-Saint-Aignan, France. E-mail: david.vaudry@univ-rouen.fr

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Abstract

PC12 cells are used to study the signaling mechanisms underlying the neurotrophic and neuroprotective activities of pituitary adenylate cyclase-activating polypeptide (PACAP) and nerve growth factor (NGF). Previous microarray experiments indicated that serpinb1a was the most induced gene after 6 h of treatment with PACAP or NGF. This study confirmed that serpinb1a is strongly activated by PACAP and NGF in a time-dependent manner with a maximum induction (~ 50-fold over control) observed after 6 h of treatment. Co-incubation with PACAP and NGF resulted in a synergistic up-regulation of serpinb1a expression (200-fold over control), suggesting that PACAP and NGF act through complementary mechanisms. Consistently, PACAP-induced serpinb1a expression was not blocked by TrkA receptor inhibition. Nevertheless, the stimulation of serpinb1a expression by PACAP and NGF was significantly reduced in the presence of extracellular signal-regulated kinase, calcineurin, protein kinase A, p38, and PI3K inhibitors, indicating that the two trophic factors share some common pathways in the regulation of serpinb1a. Finally, functional investigations conducted with siRNA revealed that serpinb1a is not involved in the effects of PACAP and NGF on PC12 cell neuritogenesis, proliferation or body cell volume but mediates their ability to block caspases 3/7 activity and to promote PC12 cell survival.

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Pituitary adenylate cyclase-activating polypeptide (PACAP) and nerve growth factor (NGF) induce a strong increase in serpinb1 a expression in PC12 cells. Functional investigations revealed that this increase in serpinb1a does not affect cell proliferation or differentiation but inhibits caspase 3 activity and promotes cell survival.

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