Autor segons l'article: Polishchuk, Aleksandra; Cilleros-Mane, Victor; Balanya-Segura, Marta; Just-Borras, Laia; Fornies-Marine, Anton; Silvera-Simon, Carolina; Tomas, Marta; El Hirchi, Meryem Jami; Hurtado, Erica; Tomas, Josep; Lanuza, Maria A
Departament: Ciències Mèdiques Bàsiques
Autor/s de la URV: Balanya Segura, Marta / Cilleros Mañé, Víctor / Hurtado Caballero, Erica / Just Borràs, Laia / Lanuza Escolano, María Angel / POLISHCHUK, ALEKSANDRA / Tomás Ferré, José Maria / Tomas Marginet, Marta / Tomàs Porres, Josep
Paraules clau: Acetylcholine release; Acetylcholine-release; Animals; B receptors; Bdnf; Bdnf protein, rat; Brain-derived neurotrophic factor; Catalytic subunit; Cyclic amp-dependent protein kinases; Dependent phosphorylation; Differential phosphorylation; Male; Modulate transmitter release; Motor endplate; Muscarinic receptors; Neuromuscular junction; Neurotrophic factor; Phosphorylation; Pk; Pka; Protein-kinase-c; Rats; Rats, wistar; Receptor, trkb; Receptors, muscarinic; Serine kinases; Signal transduction; Snap25 protein, rat; Synapsins; Synaptosomal-associated protein 25; Trk; Trkb
Resum: BackgroundProtein kinase A (PKA) enhances neurotransmission at the neuromuscular junction (NMJ), which is retrogradely regulated by nerve-induced muscle contraction to promote Acetylcholine (ACh) release through the phosphorylation of molecules involved in synaptic vesicle exocytosis (SNAP-25 and Synapsin-1). However, the molecular mechanism of the retrograde regulation of PKA subunits and its targets by BDNF/TrkB pathway and muscarinic signalling has not been demonstrated until now. At the NMJ, retrograde control is mainly associated with BDNF/TrkB signalling as muscle contraction enhances BDNF levels and controls specific kinases involved in the neurotransmission. Neurotransmission at the NMJ is also highly modulated by muscarinic receptors M1 and M2 (mAChRs), which are related to PKA and TrkB signallings. Here, we investigated the hypothesis that TrkB, in cooperation with mAChRs, regulates the activity-dependent dynamics of PKA subunits to phosphorylate SNAP-25 and Synapsin-1.MethodsTo explore this, we stimulated the rat phrenic nerve at 1Hz (30 minutes), with or without subsequent contraction (abolished by mu -conotoxin GIIIB). Pharmacological treatments were conducted with the anti-TrkB antibody clone 47/TrkB for TrkB inhibition and exogenous h-BDNF; muscarinic inhibition with Pirenzepine-dihydrochloride and Methoctramine-tetrahydrochloride for M1 and M2 mAChRs, respectively. Diaphragm protein levels and phosphorylation' changes were detected by Western blotting. Location of the target proteins was demonstrated using immunohistochemistry.ResultsWhile TrkB does not directly impact the levels of PKA catalytic subunits C alpha and C beta, it regulates PKA regulatory subunits RI alpha and RII beta, facilitating the phosphorylation of critical exocytotic targets such as SNAP-25 and Synapsin-1. Furthermore, the muscarinic receptors pathway maintains a delicate balance in this regulatory process. These findings explain the dynamic interplay of PKA subunits influenced by BDNF/TrkB signalling, M1 and M2 mAChRs pathways, that are differently regulated by pre- and postsynaptic activity, demonstrating the specific roles of the BDNF/TrkB and muscarinic receptors pathway in retrograde regulation.ConclusionThis complex molecular interplay has the relevance of interrelating two fundamental pathways in PKA-synaptic modulation: one retrograde (neurotrophic) and the other autocrine (muscarinic). This deepens the fundamental understanding of neuromuscular physiology of neurotransmission that gives plasticity to synapses and holds the potential for identifying therapeutic strategies in conditions characterized by impaired neuromuscular communication.
Grup de recerca: Unitat d'Histologia i Neurobiologia
Àrees temàtiques: Biochemistry; Biotecnología; Cell biology; Ciências biológicas i; Ciências biológicas ii; Ciências biológicas iii; Medicina i; Medicina ii; Medicina iii; Molecular biology
Accès a la llicència d'ús: https://creativecommons.org/licenses/by/3.0/es/
Adreça de correu electrònic de l'autor: mariaangel.lanuza@urv.cat; laia.just@urv.cat; josepmaria.tomas@urv.cat; victor.cilleros@alumni.urv.cat; marta.balanya@urv.cat; aleksandra.polishchuk@urv.cat; josep.tomaspo@estudiants.urv.cat; erica.hurtado@urv.cat; aleksandra.polishchuk@urv.cat; marta.tomas@urv.cat; laia.just@urv.cat; marta.balanya@urv.cat
Data d'alta del registre: 2025-01-04
Versió de l'article dipositat: info:eu-repo/semantics/publishedVersion
Enllaç font original: https://biosignaling.biomedcentral.com/articles/10.1186/s12964-024-01735-2
Referència a l'article segons font original: Cell Communication And Signaling. 22 (1): 371-
Referència de l'ítem segons les normes APA: Polishchuk, Aleksandra; Cilleros-Mane, Victor; Balanya-Segura, Marta; Just-Borras, Laia; Fornies-Marine, Anton; Silvera-Simon, Carolina; Tomas, Marta; (2024). BDNF/TrkB signalling, in cooperation with muscarinic signalling, retrogradely regulates PKA pathway to phosphorylate SNAP-25 and Synapsin-1 at the neuromuscular junction. Cell Communication And Signaling, 22(1), 371-. DOI: 10.1186/s12964-024-01735-2
URL Document de llicència: https://repositori.urv.cat/ca/proteccio-de-dades/
DOI de l'article: 10.1186/s12964-024-01735-2
Entitat: Universitat Rovira i Virgili
Any de publicació de la revista: 2024
Tipus de publicació: Journal Publications
Repositori URV