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Laboratory of Pharmacology of Neurodegeneration  

lab Prof.ssa DiLuca

 

Responsible  Prof.ssa Monica DiLuca        monica.diluca@unimi.it  

Team

Fabrizio Gardoni Professor fabrizio.gardoni@unimi.it

Elena Marcello Researcher elena.marcello@unimi.it

Nicolò Carrano PhD Student nicolo.carrano@unimi.it

Annalisa Longhi Technician annalisa.longhi@unimi.it 

Manuela Mellone Post-Doc manuela.mellone@unimi.it

Silvia Pelucchi Post-Doc silvia.pelucchi@unimi.it 

Ana Da Rocha Ribeiro PhD Student ana.darocha@unimi.it 

Tanmoy Samaddar Post-Doc tanmoy.samaddar@unimi.it

Sebastien Therin PhD Student sebastien.therin@unimi.it

Lina Vandermeulen PhD Student lina.vandermeulen@unimi.it

Elisa Zianni Technician elisa.zianni@unimi.it

 

Research Interests

Study of the molecular mechanisms involved in the pathogenesis of Alzheimer´s Disease (AD) and identification of new pharmacological targets for therapeutic intervention in AD.

Study of NMDA receptor at corticostriatal synapses in experimental models of Parkinson's disease and L-DOPA induced dyskinesia. Modulation of NMDA receptor localization and activity at corticostriatal synapses as innovative pharmacological approach for treatment of L-DOPA induced dyskinesia.

Study of the functional and structural characterization of postsynaptic density proteins of glutamatergic neurons both in physiological conditions and in neurodegenerative disorders.

Modulation of NMDA receptor localization and activity at corticostriatal synapses as innovative pharmacological approach for treatment of L-DOPA induced dyskinesia.

The research activity is related to the study of the dynamic modulation of NMDA receptor composition within the excitatory corticostriatal synapse. In particular, the goal is to study the molecular mechanisms leading to alterations of the glutamatergic synapse in the pathogenesis of Parkinson's disease and L-DOPA induced dyskinesia. The outcomes of these studies provided the basis for designing new innovative therapeutic strategies, which specifically target the dysfunction of the NMDA receptor at corticostriatal synapses. To this, we developed the use of cell-permeable TAT peptides to regulate NMDA receptor subunits availability at synaptic sites and leading to a significant decrease of L-DOPA induced dyskinesia.

Study of the molecular mechanisms involved in the pathogenesis of Alzheimer´s Disease (AD) and identification of new pharmacological targets for therapeutic intervention in AD.

Research activity is focused to the identification of molecular mechanisms involved in AD pathogenesis both by using in vitro neuronal models and post-mortem brain tissue from AD patients. These studies lead to the molecular and functional characterization of the synaptic activity of ADAM10, a member of disintegrin and metalloprotease, responsible for alpha-secretase cleavage of amyloid precursor protein (APP). Modulation of the synaptic localization and activity of ADAM10 could represent an innovative strategy to prevent pathogenic events correlated to amyloid burden and leading to the onset of AD.

Study of the functional and structural characterization of postsynaptic density proteins of glutamatergic synapses.

The main aim of this line of research is the analysis of the molecular composition of NMDA receptor complex in the excitatory postsynaptic densities and its role in the induction of synaptic plasticity events. In particular, these studies aim to characterize how scaffolding proteins and other PSD-associated proteins could affect NMDA receptor subunit trafficking and clustering in the postsynaptic compartment both in physiological conditions (i.e. synaptic plasticity) and in central nervous system disorders.

Techniques

In vitro (primary hippocampal neurons, acute of organotypic hp slices) and in vivo models of Alzheimer disease. In vitro (corticostriatal slices) and in vivo models of Parkinson disease and L-DOPA induced dyskinesia. Cell lines (COS7, HEK293, CHO). Purification of neuronal subcellular fractions (microsome, synaptosomes, postsynaptic density). In vitro and in vivo use of cell-permeable peptides. Biochemical techniques (western blotting, surface assay, internalization assay, deglycosylation assay), analysis of protein-protein interaction (pull-down, immunoprecipitation, proximity ligation assay). Molecular biology techniques (PCR, cloning, transient and stable transfections, site-directed mutagenesis). Analysis of dendritic spine morphology. Confocal imaging techniques.

 

Publications

1. Dinamarca MC, Guzzetti F, Karpova A, Lim D, Mitro N, Musardo S, Mellone M, Marcello E, Stanic J, Samaddar T, Burguière A, Caldarelli A, Genazzani AA, Perroy J, Fagni L, Canonico PL, Kreutz MR, Gardoni F, Di Luca M. Ring finger protein 10 is a novel synaptonuclear messenger encoding activation of NMDA receptors in hippocampus. Elife. 2016 Mar 15;5:e12430. doi: 10.7554/eLife.12430.

2. Stanic J, Carta M, Eberini I, Pelucchi S, Marcello E, Genazzani AA, Racca C, Mulle C, Di Luca M, Gardoni F. Rabphilin 3A retains NMDA receptors at synaptic sites through interaction with GluN2A/PSD-95 complex. Nat Commun. 2015 Dec 18;6:10181. doi: 10.1038/ncomms10181.

3. Mellone M, Stanic J, Hernandez LF, Iglesias E, Zianni E, Longhi A, Prigent A, Picconi B, Calabresi P, Hirsch EC, Obeso JA, Di Luca M, Gardoni F. NMDA receptor GluN2A/GluN2B subunit ratio as synaptic trait of levodopa-induced dyskinesias: from experimental models to patients. Front Cell Neurosci. 2015 Jul 6;9: 245. doi: 10.3389/fncel.2015.00245.

4. Pasciuto E, Ahmed T, Wahle T, Gardoni F, D'Andrea L, Pacini L, Jacquemont S, Tassone F, Balschun D, Dotti CG, Callaerts-Vegh Z, D'Hooge R, Müller UC, Di Luca M, De Strooper B, Bagni C. Dysregulated ADAM10-Mediated Processing of APP during a Critical Time Window Leads to Synaptic Deficits in Fragile X Syndrome. Neuron. 2015 Jul 15;87(2):382-98. doi: 10.1016/j.neuron.2015.06.032.

5. Gardoni F, Di Luca M. Targeting glutamatergic synapses in Parkinson's disease. Curr Opin Pharmacol. 2015 Feb; 20:24-8. doi: 10.1016/j.coph.2014.10.011.

6. Saraceno C, Marcello E, Di Marino D, Borroni B, Claeysen S, Perroy J, Padovani A, Tramontano A, Gardoni F, Di Luca M. SAP97-mediated ADAM10 trafficking from Golgi outposts depends on PKC phosphorylation. Cell Death Dis. 2014 Nov 27;5:e1547. doi: 10.1038/cddis.2014.492.

7. Marcello E, Saraceno C, Musardo S, Vara H, de la Fuente AG, Pelucchi S, Di Marino D, Borroni B, Tramontano A, Pérez-Otaño I, Padovani A, Giustetto M, Gardoni F, Di Luca M. Endocytosis of synaptic ADAM10 in neuronal plasticity and Alzheimer's disease. J Clin Invest. 2013 Jun 3; 123(6): 2523-38.

8. Vastagh C, Gardoni F, Bagetta V, Stanic J, Zianni E, Giampà C, Picconi B, Calabresi P, Di Luca M. N-methyl-D-aspartate (NMDA) receptor composition modulates dendritic spine morphology in striatal medium spiny neurons. J Biol Chem. 2012 May; 287(22): 18103-14.

9. Gardoni F, Saraceno C, Malinverno M, Marcello E, Verpelli C, Sala C, Di Luca M. The neuropeptide PACAP38 induces dendritic spine remodeling through ADAM10-N-cadherin signaling pathway. J Cell Sci. 2012 Mar; 125(Pt 6).

10. Gardoni F, Sgobio C, Pendolino V, Calabresi P, Di Luca M, Picconi B. Targeting NR2A-containing NMDA receptors reduces L-DOPA-induced dyskinesias. Neurobiol Aging. 2012 Sep; 33(9): 2138-44.

11. Marcello E, Epis R, Saraceno C, Gardoni F, Borroni B, Cattabeni F, Padovani A, Di Luca M. SAP97-mediated local trafficking is altered in Alzheimer disease patients' hippocampus. Neurobiol Aging. 2012 Feb; 33(2): 422.e1-10.

12. Malinverno M, Carta M, Epis R, Marcello E, Verpelli C, Cattabeni F, Sala C, Mulle C, Di Luca M, Gardoni F. Synaptic localization and activity of ADAM10 regulate excitatory synapses through N-cadherin cleavage. J Neurosci. 2010 Dec 1; 30(48): 16343-55.

13. Epis R, Marcello E, Gardoni F, Vastagh C, Malinverno M, Balducci C, Colombo A, Borroni B, Vara H, Dell'Agli M, Cattabeni F, Giustetto M, Borsello T, Forloni G, Padovani A, Di Luca M. Blocking ADAM10 synaptic trafficking generates a model of sporadic Alzheimer's disease. Brain. 2010 Nov; 133(11): 3323-35.

14. Marcello E, Gardoni F, Di Luca M, Pérez-Otaño I. An arginine stretch limits ADAM10 exit from the endoplasmic reticulum. J Biol Chem. 2010 Apr; 285(14): 10376-84.

15. Gardoni F, Mauceri D, Malinverno M, Polli F, Costa C, Tozzi A, Siliquini S, Picconi B, Cattabeni F, Calabresi P, Di Luca M. Decreased NR2B subunit synaptic levels cause impaired long-term potentiation but not long-term depression.  J Neurosci. 2009 Jan; 29(3): 669-77.

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