Repetitive prefrontal tDCS activates VTA dopaminergic neurons, resulting in attenuation of Alzheimer’s Disease-like deficits in Tg2576 mice Maria Luisa De Paolis, Gilda Loffredo, Paraskevi Krashia, Livia La Barbera, Annalisa Nobili, Emma Cauzzi, Lucy Babicola, Matteo Di Segni, Roberto Coccurello, Stefano Puglisi-Allegra, Emanuele Claudio Latagliata, Marcello D’Amelio Alzheimer S Research and Therapy, 2025 BACKGROUND: Emerging evidence implicates early dysfunction of dopaminergic neurons in the Ventral Tegmental Area (VTA) as a key contributor to Alzheimer's Disease (AD) pathophysiology. Specifically, the VTA dopaminergic neurodegeneration and the consequent reduction of dopamine (DA) in mesocorticolimbic targets are associated with the onset of cognitive impairments and neuropsychiatric-like manifestations in AD animal models. Moreover, decreased midbrain volume and functional VTA disconnection are identified as predictors of accelerated progression from Mild Cognitive Impairment to AD-dementia in clinical populations. Given these findings, interventions capable of directly modulating VTA activity and augmenting DA release, despite the ongoing neurodegeneration, may hold therapeutic potential for mitigating DA-related deficits in AD. This study aims at evaluating the therapeutic potential of prefrontal transcranial Direct Current Stimulation (tDCS) in the Tg2576 mouse model of AD, exhibiting early VTA dopaminergic neurodegeneration. METHODS: Repeated tDCS was applied to assess its ability to activate VTA DA neurons. We also evaluated tDCS effects on synaptic plasticity, cognitive and non-cognitive behaviours and AD-related pathology. Hippocampal DA release and Nucleus Accumbens (NAc) DA transporter (DAT) expression were measured. With immunohistochemistry we examined microglial density and morphological complexity at different disease stages. Additionally, intracellular amyloid-β (Aβ) levels and plaque burden were evaluated to determine the impact of tDCS on AD pathology. RESULTS: Prefrontal tDCS enhanced the activity of VTA dopaminergic neurons, leading to increased hippocampal DA release and higher DAT levels in the NAc. The enhanced DA outflow is associated with restored CA3-CA1 synaptic plasticity and improvements in recognition memory and motivational behaviours. tDCS reduced microglial numbers and morphological complexity in Tg2576 mice at both pre-plaque stage (7-months) and at an advanced stage characterized by plaque accumulation (12-months). Notably, tDCS also decreased Aβ plaque burden, although no changes in intracellular Aβ levels were observed in younger Tg2576 mice. CONCLUSIONS: These findings highlight the multifaceted therapeutic potential of prefrontal tDCS in targeting key AD pathophysiological hallmarks, including dopaminergic dysfunction, synaptic impairments, neuroinflammation and plaque deposition. As a non-invasive neuromodulatory approach, prefrontal tDCS emerges as a promising early intervention strategy to complement existing AD treatments, with the potential to improve patient outcomes and quality of life.
Midbrain degeneration triggers astrocyte reactivity and tau pathology in experimental Alzheimer’s Disease Livia La Barbera, Paraskevi Krashia, Gilda Loffredo, Emma Cauzzi, Maria Luisa De Paolis, Martina Montanari, Luana Saba, Elena Spoleti, Serena Ficchì, Claudio Zaccone, Marco De Bardi, Claudia Palazzo, Ramona Marino, Emanuele Claudio Latagliata, Stefano Puglisi-Allegra, Giovanna Borsellino, Flavio Keller, Luisa Lo Iacono, Maria Teresa Viscomi, Annalisa Nobili, Marcello D’Amelio Molecular Neurodegeneration, 2025 Background Smaller midbrain volumes predict Alzheimer’s Disease (AD) progression and faster conversion from Mild Cognitive Impairment (MCI) to dementia. Along with this, various midbrain-target areas are characterized by neuroinflammation since the MCI stage. The concomitance of neuroinflammation, Αβ and tau appears to be a strong predictor for conversion from MCI to dementia. Yet, how midbrain degeneration could cause disease progression, and what mechanisms are involved in triggering neuroinflammation in midbrain-target areas such as the hippocampus remain unexplored. Methods Using adult C57BL/6N mice we generated a new mouse model carrying lesions in three midbrain nuclei, the dopaminergic Ventral Tegmental Area (VTA) and Substantia Nigra pars compacta (SNpc) and the serotonergic Interpeduncular Nucleus (IPN), to evaluate the consequences of dopamine and serotonin deprivation in midbrain-target areas. We characterized this model by performing stereological cell counts, analysis of monoaminergic fibers, monoamine levels, electrophysiology and behavioral tests. We then assessed hippocampal neuroinflammation by analyzing glia cell count, changes in morphology, NLRP3 inflammasome activation and cytokine levels, and microglia transcriptional profiling. In a separate set of experiments, we induced experimental midbrain lesion in Tg2576 transgenic mice overexpressing the Swedish mutant amyloid precursor protein, to evaluate the effect of monoamine deprivation on the hippocampus in concomitance with amyloid-β (Aβ) accumulation. The lesion performed in Tg2576 mice, as opposed to that in C57BL/6N mice, provides valuable insights into how neuroinflammation is influenced by Aβ accumulation versus the exclusive impact of impaired monoaminergic signaling. Results The concomitant depletion of dopaminergic and serotonergic inputs within the hippocampus of C57BL/6N mice provokes a pronounced activation of microglia via the NLRP3-inflammasome pathway, accompanied by increased IL-1β expression. Pharmacological intervention with either dopaminergic (L-DOPA or A68930) or serotonergic (fluoxetine) agents abrogates this neuroinflammatory response. In the Tg2576 transgenic mouse model of amyloid pathology, which exhibits progressive Aβ deposition, superimposed midbrain degeneration markedly amplifies AD-like neuropathology. This includes exacerbation of microglial reactivity, robust astrocyte response, precocious Aβ plaque burden, and induction of pathological tau hyperphosphorylation. Notably, administration of L-DOPA or fluoxetine significantly attenuates both the astrocyte reactivity and tau hyperphosphorylation in the lesioned Tg2576 cohort. Conclusions These results highlight the pivotal role of midbrain damage for the amplification of neuroinflammatory cascades and AD pathology. Moreover, they offer mechanistic insight into the faster progression to dementia in patients with midbrain deficits. By translating these findings into clinical practice, we can advance towards a precision medicine approach in disease management.
In situ stoichiometry amounts of p62 and poly-ubiquitin exceed the increase of alpha-synuclein during degeneration of catecholamine cells induced by autophagy inhibition in vitro Paola Lenzi, Gloria Lazzeri, Michela Ferrucci, Carla Letizia Busceti, Stefano Puglisi-Allegra, Francesco Fornai Journal of Neural Transmission, 2024 Neurodegenerative disorders are typically featured by the occurrence of neuronal inclusions. In the case of Parkinson’s disease (PD) these correspond to Lewy bodies (LBs), which are routinely defined as proteinaceous inclusions composed of alpha-synuclein (alpha-syn). In turn, alpha-syn is considered to be the key protein in producing PD and fostering its progression. Recent studies challenged such a concept and emphasized the occurrence of other proteins such as p62 and poly-ubiquitin (Poly-ub) in the composition of LBs, which are also composed of large amounts of tubulo-vesicular structures. All these components, which accumulate within the cytosol of affected neurons in PD, may be the consequence of a dysfunction of major clearing pathways. In fact, autophagy-related systems are constantly impaired in inherited PD and genetic models of PD. The present study was designed to validate whether a pharmacological inhibition of autophagy within catecholamine cells produces cell damage and accumulation of specific proteins and tubulo-vesicular structures. The stoichiometry counts of single proteins, which accumulate within catecholamine neurons was carried out along with the area of tubulo-vesicular structures. In these experimental conditions p62 and Poly-ub accumulation exceeded at large the amounts of alpha-syn. In those areas where Poly-ub and p62 were highly expressed, tubulo-vesicular structures were highly represented compared with surrounding cytosol. The present study confirms new vistas about LBs composition and lends substance to the scenario that autophagy inhibition rather than a single protein dysfunction as key determinant of PD.
Methamphetamine-Induced Blood Pressure Sensitization Correlates with Morphological Alterations within A1/C1 Catecholamine Neurons Carla Letizia Busceti, Domenico Bucci, Antonio Damato, Massimiliano De Lucia, Eleonora Venturini, Michela Ferrucci, Gloria Lazzeri, Stefano Puglisi-Allegra, Mariarosaria Scioli, Albino Carrizzo, Ferdinando Nicoletti, Carmine Vecchione, Francesco Fornai International Journal of Molecular Sciences, 2024 Methamphetamine (METH) is a drug of abuse, which induces behavioral sensitization following repeated doses. Since METH alters blood pressure, in the present study we assessed whether systolic and diastolic blood pressure (SBP and DBP, respectively) are sensitized as well. In this context, we investigated whether alterations develop within A1/C1 neurons in the vasomotor center. C57Bl/6J male mice were administered METH (5 mg/kg, daily for 5 consecutive days). Blood pressure was measured by tail-cuff plethysmography. We found a sensitized response both to SBP and DBP, along with a significant decrease of catecholamine neurons within A1/C1 (both in the rostral and caudal ventrolateral medulla), while no changes were detected in glutamic acid decarboxylase. The decrease of catecholamine neurons was neither associated with the appearance of degeneration-related marker Fluoro-Jade B nor with altered expression of α-synuclein. Rather, it was associated with reduced free radicals and phospho-cJun and increased heat shock protein-70 and p62/sequestosome within A1/C1 cells. Blood pressure sensitization was not associated with altered arterial reactivity. These data indicate that reiterated METH administration may increase blood pressure persistently and may predispose to an increased cardiovascular response to METH. These data may be relevant to explain cardiovascular events following METH administration and stressful conditions.
RAB32 mutation in Parkinson's disease [2] Edoardo Monfrini, Raffaella Minardi, Franco Valzania, Giovanna Calandra-Buonaura, Paola Mandich, Alessio Di Fonzo, Valerio Carelli, Andrea Gaudio, Agata Fant, Ilaria Palmieri, Valentina Fioravanti, Francesco Cavallieri, Francesca Di Biasio, Alessia Fiorentino, Manuela Vecchi, Paola Origone, Sara Trova, Giovanni Rizzo, Giulia Giannini, Stefano Gambardella, Micol Avenali, Stefano Gustincich, Pietro Cortelli, Fabio Blandini, Enza Maria Valente Lancet Neurology, 2024
Methamphetamine Increases Tubulo-Vesicular Areas While Dissipating Proteins from Vesicles Involved in Cell Clearance Gloria Lazzeri, Paola Lenzi, Carla L. Busceti, Stefano Puglisi-Allegra, Michela Ferrucci, Francesco Fornai International Journal of Molecular Sciences, 2024 Cytopathology induced by methamphetamine (METH) is reminiscent of degenerative disorders such as Parkinson’s disease, and it is characterized by membrane organelles arranged in tubulo-vesicular structures. These areas, appearing as clusters of vesicles, have never been defined concerning the presence of specific organelles. Therefore, the present study aimed to identify the relative and absolute area of specific membrane-bound organelles following a moderate dose (100 µM) of METH administered to catecholamine-containing PC12 cells. Organelles and antigens were detected by immunofluorescence, and they were further quantified by plain electron microscopy and in situ stoichiometry. This analysis indicated an increase in autophagosomes and damaged mitochondria along with a decrease in lysosomes and healthy mitochondria. Following METH, a severe dissipation of hallmark proteins from their own vesicles was measured. In fact, the amounts of LC3 and p62 were reduced within autophagy vacuoles compared with the whole cytosol. Similarly, LAMP1 and Cathepsin-D within lysosomes were reduced. These findings suggest a loss of compartmentalization and confirm a decrease in the competence of cell clearing organelles during catecholamine degeneration. Such cell entropy is consistent with a loss of energy stores, which routinely govern appropriate subcellular compartmentalization.
Combined light and electron microscopy (CLEM) to quantify methamphetamine-induced alpha-synuclein-related pathology Michela Ferrucci, Paola Lenzi, Gloria Lazzeri, Carla L. Busceti, Alessandro Frati, Stefano Puglisi-Allegra, Francesco Fornai Journal of Neural Transmission, 2024 Methamphetamine (METH) produces a cytopathology, which is rather specific within catecholamine neurons both in vitro and ex vivo, in animal models and chronic METH abusers. This led some authors to postulate a sort of parallelism between METH cytopathology and cell damage in Parkinson’s disease (PD). In fact, METH increases and aggregates alpha-syn proto-fibrils along with producing spreading of alpha-syn. Although alpha-syn is considered to be the major component of aggregates and inclusions developing within diseased catecholamine neurons including classic Lewy body (LB), at present, no study provided a quantitative assessment of this protein in situ, neither following METH nor in LB occurring in PD. Similarly, no study addressed the quantitative comparison between occurrence of alpha-syn and other key proteins and no investigation measured the protein compared with non-protein structure within catecholamine cytopathology. Therefore, the present study addresses these issues using an oversimplified model consisting of a catecholamine cell line where the novel approach of combined light and electron microscopy (CLEM) was used measuring the amount of alpha-syn, which is lower compared with p62 or poly-ubiquitin within pathological cell domains. The scenario provided by electron microscopy reveals unexpected findings, which are similar to those recently described in the pathology of PD featuring packing of autophagosome-like vesicles and key proteins shuttling autophagy substrates. Remarkably, small seed-like areas, densely packed with p62 molecules attached to poly-ubiquitin within wide vesicular domains occurred. The present data shed new light about quantitative morphometry of catecholamine cell damage in PD and within the addicted brain.
Pain in Parkinson's disease: a neuroanatomy-based approach Domiziana Nardelli, Francesco Gambioli, Maria Ilenia De Bartolo, Romina Mancinelli, Francesca Biagioni, Simone Carotti, Emma Falato, Giorgio Leodori, Stefano Puglisi-Allegra, Giorgio Vivacqua, Francesco Fornai Brain Communications, 2024
Biological determinants of blood-based cytokines in the Alzheimer's disease clinical continuum Alessandro Galgani, Andrea Vergallo, Nicole Campese, Francesco Lombardo, Nicola Pavese, Lucia Petrozzi, Annalisa LoGerfo, Maria Franzini, Denise Cecchetti, Stefano Puglisi‐Allegra, Carla L. Busceti, Gabriele Siciliano, Gloria Tognoni, Filippo Baldacci, Simone Lista, Harald Hampel, Francesco Fornai, Filippo S. Giorgi Journal of Neurochemistry, 2022
Neuroprotective effects of curcumin in methamphetamine-induced toxicity Larisa Ryskalin, Stefano Puglisi-Allegra, Gloria Lazzeri, Francesca Biagioni, Carla L. Busceti, Linda Balestrini, Andrea Fornasiero, Stefano Leone, Elena Pompili, Michela Ferrucci, Francesco Fornai Molecules, 2021
From Traumatic Childhood to Cocaine Abuse: The Critical Function of the Immune System Luisa Lo Iacono, Clarissa Catale, Alessandro Martini, Alessandro Valzania, Maria Teresa Viscomi, Valerio Chiurchiù, Ezia Guatteo, Silvia Bussone, Fabiana Perrone, Paola Di Sabato, Eleonora Aricò, Alberto D’Argenio, Alfonso Troisi, Nicola B. Mercuri, Mauro Maccarrone, Stefano Puglisi-Allegra, Pietro Casella, Valeria Carola Biological Psychiatry, 2018
Cerebellar BDNF Promotes Exploration and Seeking for Novelty Daniela Laricchiuta, Diego Andolina, Francesco Angelucci, Francesca Gelfo, Erica Berretta, Stefano Puglisi-Allegra, Laura Petrosini International Journal of Neuropsychopharmacology, 2018
A new therapy prevents intellectual disability in mouse with phenylketonuria Tiziana Pascucci, Luigia Rossi, Marco Colamartino, Claudia Gabucci, Claudia Carducci, Alessandro Valzania, Valeria Sasso, Noemi Bigini, Francesca Pierigè, Maria Teresa Viscomi, Rossella Ventura, Simona Cabib, Mauro Magnani, Stefano Puglisi-Allegra, Vincenzo Leuzzi Molecular Genetics and Metabolism, 2018
miR-34b/c Regulates Wnt1 and Enhances Mesencephalic Dopaminergic Neuron Differentiation Roberto De Gregorio, Salvatore Pulcrano, Claudia De Sanctis, Floriana Volpicelli, Ezia Guatteo, Lars von Oerthel, Emanuele Claudio Latagliata, Roberta Esposito, Rosa Maria Piscitelli, Carla Perrone-Capano, Valerio Costa, Dario Greco, Stefano Puglisi-Allegra, Marten P. Smidt, Umberto di Porzio, Massimiliano Caiazzo, Nicola Biagio Mercuri, Meng Li, Gian Carlo Bellenchi Stem Cell Reports, 2018
Dopamine neuronal loss contributes to memory and reward dysfunction in a model of Alzheimer's disease Annalisa Nobili, Emanuele Claudio Latagliata, Maria Teresa Viscomi, Virve Cavallucci, Debora Cutuli, Giacomo Giacovazzo, Paraskevi Krashia, Francesca Romana Rizzo, Ramona Marino, Mauro Federici, Paola De Bartolo, Daniela Aversa, Maria Concetta Dell’Acqua, Alberto Cordella, Marco Sancandi, Flavio Keller, Laura Petrosini, Stefano Puglisi-Allegra, Nicola Biagio Mercuri, Roberto Coccurello, Nicola Berretta, Marcello D’Amelio Nature Communications, 2017
GABA content within the ventromedial prefrontal cortex is related to trait anxiety Stefano Delli Pizzi, Caterina Padulo, Alfredo Brancucci, Giovanna Bubbico, Richard A. Edden, Antonio Ferretti, Raffaella Franciotti, Valerio Manippa, Daniele Marzoli, Marco Onofrj, Gianna Sepede, Armando Tartaro, Luca Tommasi, Stefano Puglisi-Allegra, Laura Bonanni Social Cognitive and Affective Neuroscience, 2016
Alpha-Synuclein Produces Early Behavioral Alterations via Striatal Cholinergic Synaptic Dysfunction by Interacting With GluN2D N-Methyl-D-Aspartate Receptor Subunit Alessandro Tozzi, Antonio de Iure, Vincenza Bagetta, Michela Tantucci, Valentina Durante, Ana Quiroga-Varela, Cinzia Costa, Massimiliano Di Filippo, Veronica Ghiglieri, Emanuele Claudio Latagliata, Michal Wegrzynowicz, Mickael Decressac, Carmela Giampà, Jeffrey W. Dalley, Jing Xia, Fabrizio Gardoni, Manuela Mellone, Omar Mukhtar El-Agnaf, Mustafa Taleb Ardah, Stefano Puglisi-Allegra, Anders Björklund, Maria Grazia Spillantini, Barbara Picconi, Paolo Calabresi Biological Psychiatry, 2016
Adversity in childhood and depression: Linked through SIRT1 L Lo Iacono, F Visco-Comandini, A Valzania, M T Viscomi, M Coviello, A Giampà, L Roscini, E Bisicchia, A Siracusano, A Troisi, S Puglisi-Allegra, V Carola Translational Psychiatry, 2015
Family-based association study of ITGB3 in autism spectrum disorder and its endophenotypes Valerio Napolioni, Federica Lombardi, Roberto Sacco, Paolo Curatolo, Barbara Manzi, Riccardo Alessandrelli, Roberto Militerni, Carmela Bravaccio, Carlo Lenti, Monica Saccani, Cindy Schneider, Raun Melmed, Tiziana Pascucci, Stefano Puglisi-Allegra, Karl-Ludvig Reichelt, Francis Rousseau, Patricia Lewin, Antonio M Persico European Journal of Human Genetics, 2011
Clinical, Morphological, and Biochemical Correlates of Head Circumference in Autism Roberto Sacco, Roberto Militerni, Alessandro Frolli, Carmela Bravaccio, Antonella Gritti, Maurizio Elia, Paolo Curatolo, Barbara Manzi, Simona Trillo, Carlo Lenti, Monica Saccani, Cindy Schneider, Raun Melmed, Karl-Ludvig Reichelt, Tiziana Pascucci, Stefano Puglisi-Allegra, Antonio M. Persico Biological Psychiatry, 2007
Enhanced APOE2 transmission rates in families with autistic probands A. M. Persico, L. D'Agruma, L. Zelante, R. Militerni, C. Bravaccio, C. Schneider, R. Melmed, S. Trillo, F. Montecchi, M. Elia, M. Palermo, D. Rabinowitz, T. Pascucci, S. Puglisi-Allegra, K.-L. Reichelt, L. Muscarella, V. Guarnieri, J.-M. Melgari, M. Conciatori, F. Keller Psychiatric Genetics, 2004
Association between the HOXA1 A218G polymorphism and increased head circumference in patients with autism Monica Conciatori, Christopher J Stodgell, Susan L Hyman, Melanie O'Bara, Roberto Militerni, Carmela Bravaccio, Simona Trillo, Francesco Montecchi, Cindy Schneider, Raun Melmed, Maurizio Elia, Lori Crawford, Sarah J Spence, Lucianna Muscarella, Vito Guarnieri, Leonardo D'Agruma, Alessandro Quattrone, Leopoldo Zelante, Daniel Rabinowitz, Tiziana Pascucci, Stefano Puglisi-Allegra, Karl-Ludvig Reichelt, Patricia M Rodier, Antonio M Persico Biological Psychiatry, 2004
The phenylketonuria mouse model: A meeting review J David McDonald, Maria Andriolo, Francesco Calı̀, Mario Mirisola, Stefano Puglisi-Allegra, Valentino Romano, Christineh N Sarkissian, Carolyn B Smith Molecular Genetics and Metabolism, 2002
Reelin gene alleles and haplotypes as a factor predisposing to autistic disorder A M Persico, L D'Agruma, N Maiorano, A Totaro, R Militerni, C Bravaccio, T H Wassink, C Schneider, R Melmed, S Trillo, F Montecchi, M Palermo, T Pascucci, S Puglisi-Allegra, K-L Reichelt, M Conciatori, R Marino, C C Quattrocchi, A Baldi, L Zelante, P Gasparini, F Keller Molecular Psychiatry, 2001
Adenosine deaminase alleles and autistic disorder: Case-control and family-based association studies Antonio M. Persico, Roberto Militerni, Carmela Bravaccio, Cindy Schneider, Raun Melmed, Simona Trillo, Francesco Montecchi, Mark T. Palermo, Tiziana Pascucci, Stefano Puglisi-Allegra, Karl-Ludvig Reichelt, Monica Conciatori, Alfonso Baldi, Flavio Keller American Journal of Medical Genetics Neuropsychiatric Genetics, 2000
Behavioural data on dermorphins in mice Stefano Puglisi-Allegra, Claudio Castellano, Umberto Filibeck, Alberto Oliverio, Pietro Melchiorri European Journal of Pharmacology, 1982
