Articles by Rosa Gómez-Villafuertes in JoVE
Live Imaging Followed by Single Cell Tracking to Monitor Cell Biology and the Lineage Progression of Multiple Neural Populations Rosa Gómez-Villafuertes*1,2,3, Lucía Paniagua-Herranz*1,2,3, Sergio Gascon*4,5, David de Agustín-Durán1,2,3, María de la O Ferreras1,2,3, Juan Carlos Gil-Redondo1,2,3, María José Queipo1,2,3, Aida Menendez-Mendez1,2,3, Ráquel Pérez-Sen1,2,3, Esmerilda G. Delicado1,2,3, Javier Gualix1,2,3, Marcos R. Costa6, Timm Schroeder7, María Teresa Miras-Portugal1,2,3, Felipe Ortega1,2,3 1Biochemistry and Molecular Biology Department, Faculty of Veterinary medicine, Complutense University, 2University Institute for Neurochemistry Research (IUIN), 3Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC), 4Institute of Stem Cell Research, Helmholtz Center Munich, Neuherberg/Munich, Germany Physiological Genomics, Biomedical Center, Ludwig-Maximilians University Munich, 5Toxicology and Pharmacology Department, Faculty of Veterinary medicine, Complutense University, 6Brain Institute, Federal University of Rio Grande do Norte, 7Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zurich A robust protocol to monitor neural populations by time-lapse video-microscopy followed by software-based post-processing is described. This method represents a powerful tool to identify biological events in a selected population during live imaging experiments.
Other articles by Rosa Gómez-Villafuertes on PubMed
Role of P2X7 and P2Y2 Receptors on α-secretase-dependent APP Processing: Control of Amyloid Plaques Formation "in Vivo" by P2X7 Receptor Computational and Structural Biotechnology Journal. | Pubmed ID: 25848496 Amyloid precursor protein (APP) is expressed in a large variety of neural and non-neural cells. The balance between non-pathogenic and pathologic forms of APP processing, mediated by α-secretase and β-secretase respectively, remains a crucial step to understand β-amyloid, Aβ42 peptide, formation and aggregation that are at the origin of the senile plaques in the brain, a characteristic hallmark of Alzheimer's disease (AD). In Neuro-2a, a neuroblastoma cell line that constitutively expresses APP, activation of the P2X7 receptor leads to reduction of α-secretase activity, the opposite effect being obtained by P2Y2 receptor activation. The in vivo approach was made possible by the use of J20 mice, a transgenic mouse model of familial Alzheimer's disease (FAD) expressing human APP mutant protein. This animal exhibits prominent amyloid plaques by six months of age. In vivo inhibition of the P2X7 receptor induced a significant decrease in the number and size of hippocampal amyloid plaques. This reduction is mediated by an increase in the proteolytic processing of APP through α-secretase activity, which correlates with an increase in the phosphorylated form of GSK-3, a less active form of this enzyme. The in vivo findings corroborate the therapeutic potential of P2X7 antagonists in the treatment of FAD.
Age-related Nuclear Translocation of P2X6 Subunit Modifies Splicing Activity Interacting with Splicing Factor 3A1 PloS One. | Pubmed ID: 25874565 P2X receptors are ligand-gated ion channels sensitive to extracellular nucleotides formed by the assembling of three equal or different P2X subunits. In this work we report, for the first time, the accumulation of the P2X6 subunit inside the nucleus of hippocampal neurons in an age-dependent way. This location is favored by its anchorage to endoplasmic reticulum through its N-terminal domain. The extracellular domain of P2X6 subunit is the key to reach the nucleus, where it presents a speckled distribution pattern and is retained by interaction with the nuclear envelope protein spectrin α2. The in vivo results showed that, once inside the nucleus, P2X6 subunit interacts with the splicing factor 3A1, which ultimately results in a reduction of the mRNA splicing activity. Our data provide new insights into post-transcriptional regulation of mRNA splicing, describing a novel mechanism that could explain why this process is sensitive to changes that occur with age.
Expression and Function of the Metabotropic Purinergic P2Y Receptor Family in Experimental Seizure Models and Patients with Drug-refractory Epilepsy Epilepsia. | Pubmed ID: 28733972 ATP is released into the extracellular space during pathologic processes including increased neuronal firing. Once released, ATP acts on P2 receptors including ionotropic P2X and metabotropic P2Y receptors, resulting in changes to glial function and neuronal network excitability. Evidence suggests an involvement of P2Y receptors in the pathogenesis of epilepsy, but there has been no systematic effort to characterize the expression and function of the P2Y receptor family during seizures and in experimental and human epilepsy.