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Identification of Proteins Regulated by PACAP in PC12 Cells by 2D Gel Electrophoresis Coupled to Mass Spectrometry

Alexis Lebon 1 Damien Seyer 2 Pascal Cosette 3 Laurent Coquet 4 Thierry Jouenne 4 Philippe Chan 5 Jérôme Leprince 5, 6, 7 Alain Fournier 8 Hubert Vaudry 5 Bruno Gonzalez 9 David Vaudry 6
1 PRIMACEN - Plate-Forme de Recherche en Imagerie Cellulaire de Haute-Normandie
IRIB - Institute for Research and Innovation in Biomedicine, UNIROUEN - Université de Rouen Normandie, HeRacLeS - High-tech Research Infrastructures for Life Sciences
2 ERRMECe - Equipe de recherche sur les relations matrice extracellulaire-cellules
3 PBS - Polymères Biopolymères Surfaces
4 PBM - Polymères, biopolymères, membranes
5 Neuroendocrinologie cellulaire et moléculaire
6 DC2N - Différenciation et communication neuronale et neuroendocrine
7 DC2N - Différenciation et communication neuronale et neuroendocrine
8 MONTREAL HEART INSTITUTE - Montreal Heart Institute
9 Néovasc - Endothélium microcirculatoire cérébral et lésions du système nerveux central au cours du développement
Abstract : The rat pheochromocytoma PC12 cell line has been widely used as a model to study neuronal differentiation. In particular, after serum depletion, PC12 cells stop to proliferate and undergo apoptosis. Under such conditions, treatment with pituitary adenylate cyclase-activating polypeptide (PACAP) promotes cell survival and induces neurite outgrowth. The identification of the proteins regulated by PACAP in PC12 cells under apoptotic conditions should provide valuable information concerning the mechanisms controlling neuronal cell survival and differentiation. To this aim, PC12 cells cultured in serum-free medium were treated with PACAP (10(-7) M), proteins were extracted, separated by two-dimensional gel electrophoresis (2-DE), and identified by MALDI-ToF mass spectrometry. The comparison between 16 2-DE maps led to the characterization of 110 proteins regulated by PACAP among which 22 have been identified by automatic query of the Mascot, Aldente, and Profound servers with the ProGeR-CDD database. Seventy-six percent of these proteins, including the p17 subunit of caspase-3, the heat shock protein hsp60, and the GTPase ran were found to be repressed whereas the others notably hsp27, tubulin beta-5, and calmodulin were overexpressed. Investigation of the putative functions indicated that some of the proteins regulated by PACAP and identified in the present article could control cell survival or differentiation.
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https://hal-normandie-univ.archives-ouvertes.fr/hal-01962805
Contributor : Jérôme Leprince Connect in order to contact the contributor
Submitted on : Thursday, December 20, 2018 - 6:56:04 PM
Last modification on : Tuesday, May 10, 2022 - 2:48:02 PM

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COMUE-NORMANDIE | CNRS | UNIV-CERGY | INSA-ROUEN | UNIROUEN | DC2N | INSA-GROUPE | PBS | ENSICAEN | UNILEHAVRE | UNICAEN | ERRMECE | I-MAT | CY-TECH-SE | INC-CNRS

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Alexis Lebon, Damien Seyer, Pascal Cosette, Laurent Coquet, Thierry Jouenne, et al.. Identification of Proteins Regulated by PACAP in PC12 Cells by 2D Gel Electrophoresis Coupled to Mass Spectrometry. Annals of the New York Academy of Sciences, Wiley, 2006, 1070 (1), pp.380-387. ⟨10.1196/annals.1317.049⟩. ⟨hal-01962805⟩

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