title: UTP affects the Schwannoma cell line proteome through P2Y receptors leading to cytoskeletal reorganisation.
abstract: Glial cells in the peripheral nervous system, such as Schwann cells, respond to nucleotides, which play an important role in axonal regeneration and myelination. Metabotropic P2Y receptor agonists are promising therapeutic molecules for peripheral neuropathies. Nevertheless, the proteomic mechanisms involved in nucleotide action on Schwann cells remain unknown. Here, we studied intracellular protein changes in RT4-D6P2T Schwann cells after treatment with nucleotides and Nucleo CMP Forte (CMPF), a nucleotide-based drug. After treatment with CMPF, 2-D DIGE revealed 11 differential gel spots, which were all upregulated. Among these, six different proteins were identified by MS. Some of these proteins are involved in actin remodelling (actin-related protein, Arp3), membrane vesicle transport (Rab GDP dissociation inhibitor β, Rab GDI), and the endoplasmic reticulum stress response (protein disulfide isomerase A3, PDI), which are hallmarks of a possible P2Y receptor signalling pathway. Expression of P2Y receptors in RT4-D6P2T cells was demonstrated by RT-PCR and a transient elevation of intracellular calcium measured in response to UTP. Actin reorganisation was visualized after UTP treatment using phalloidin-FITC staining and was blocked by the P2Y antagonist suramin, which also inhibited Arp3, Rab GDI, and PDI protein upregulation. Our data indicate that extracellular UTP interacts with Schwann P2Y receptors and activates molecular machinery that induces changes in the glial cell cytoskeleton.
authors: Faculty of Medicine and Health Sciences, Universitat Internacional de Catalunya, Josep Trueta s/n, Sant Cugat del Vallès, Spain.
authors: Tánia
authors: T
authors: Martiáñez
authors: Montserrat
authors: M
authors: Carrascal
authors: Aloa
authors: A
authors: Lamarca
authors: Mònica
authors: M
authors: Segura
authors: Núria
authors: N
authors: Durany
authors: Roser
authors: R
authors: Masgrau
authors: Joaquín
authors: J
authors: Abian
authors: Alejandro
authors: A
authors: Gella
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003568:Cytidine Monophosphate / Q000494:pharmacology; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression; D009442:Neurilemmoma; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D058912:Purinergic P2Y Receptor Agonists / Q000494:pharmacology*; D051381:Rats; D058470:Receptors, Purinergic P2Y / Q000235:genetics / Q000378:metabolism*; D057825:Two-Dimensional Difference Gel Electrophoresis; D015854:Up-Regulation / Q000187:drug effects; D014544:Uridine Triphosphate / Q000494:pharmacology*
chemical_list: D003598:Cytoskeletal Proteins; D020543:Proteome; D058912:Purinergic P2Y Receptor Agonists; D058470:Receptors, Purinergic P2Y; D003568:Cytidine Monophosphate; D014544:Uridine Triphosphate