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Proteomic dissection of substrate nanotopography-sensitive mechanotransductive signalling hubs that foster neuronal differentiation.

ABSTRACT: Neuronal cells are competent in precisely sensing nanotopographical features of their microenvironment and the intrinsic information impacts on neuronal functioning and differentiation “interpreted” by mechanotransductive processes. Attempts to influence and direct neuronal differentiation behaviour by engineering substrates that mimic appropriate extracellular matrix (ECM) topographies are hampered by the difficulty since many details of mechanosensing/-transduction are still elusive. Introducing omics methods into these biomaterial approaches has the potential to provide a deeper insight into the molecular processes and signalling cascades underlying mechanosensing/-transduction but their exigence in cellular material is often opposed by technical limitations of the biomaterial fabrication methods. Supersonic cluster beam deposition (SCBD) of zirconia nanoparticles allows instead the realisation of large macroscopic areas with biocompatible nanostructured films equipped with controllable and reproducible ECM-like nanoroughness that have been recently shown to foster neuron differentiation and maturation. Exploiting this capacity of SCBD, we challenged mechanosensing/-transduction and differentiative behaviour of neuron-like PC12 cells interacting with diverse nanotopographies and/or by changing their biomechanical status, to analyse their phosphoproteomic profiles in these settings. Versatile proteins were found to be affected that can be associated to significant processes along the mechanotransductive signal sequence, i.e. cell/cell interaction, glycocalyx and ECM, membrane/f-actin linkage and integrin activation, cell/substrate interaction, IAC, actomyosin organisation/cellular mechanics, nuclear organisation and transcription. The phosphoproteomic data suggested furthermore an involvement of ILK, mTor, Wnt and calcium signalling in these nanotopography- and/or cell mechanics-related processes. Altogether, potential nanotopography-sensitive, mechanotransductive signalling hubs participating to neuronal differentiation were dissected.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Cell Culture

SUBMITTER: gabriella tedeschi

LAB HEAD: Gabriella Tedeschi

PROVIDER: PXD007644 | Pride | 2018-01-24

REPOSITORIES: Pride

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Publications

Methionine supplementation stimulates mitochondrial respiration.

Tripodi Farida F Castoldi Andrea A Nicastro Raffaele R Reghellin Veronica V Lombardi Linda L Airoldi Cristina C Falletta Ermelinda E Maffioli Elisa E Scarcia Pasquale P Palmieri Luigi L Alberghina Lilia L Agrimi Gennaro G Tedeschi Gabriella G Coccetti Paola P

Biochimica et biophysica acta. Molecular cell research 20181002 12

Mitochondria play essential metabolic functions in eukaryotes. Although their major role is the generation of energy in the form of ATP, they are also involved in maintenance of cellular redox state, conversion and biosynthesis of metabolites and signal transduction. Most mitochondrial functions are conserved in eukaryotic systems and mitochondrial dysfunctions trigger several human diseases. By using multi-omics approach, we investigate the effect of methionine supplementation on yeast cellular ...[more]

PMID: 30290237

Publication: 1/2

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