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30 results on '"Franchino, Claudio"'

1. Enhancing the Study of Quantal Exocytotic Events: Combining Diamond Multi-Electrode Arrays with Amperometric PEak Analysis (APE) an Automated Analysis Code

Author

Tomagra, Giulia, Re, Alice, Varzi, Veronica, Aprà, Pietro, Britel, Adam, Franchino, Claudio, Sturari, Sofia, Amine, Nour-Hanne, Westerink, Remco H. S., Carabelli, Valentina, and Picollo, Federico

Subjects
Physics - Biological Physics
Abstract

MicroGraphited-Diamond-Multi Electrode Arrays ({\mu}G-D-MEAs) can be successfully used to reveal, in real time, quantal exocytotic events occurring from many individual neurosecretory cells and/or from many neurons within a network. As {\mu}G-D-MEAs arrays are patterned with up to 16 sensing microelectrodes, each of them recording large amounts of data revealing the exocytotic activity, the aim of this work was to support an adequate analysis code to speed up the signal detection. The cutting-edge technology of microGraphited-Diamond-Multi Electrode Arrays ({\mu}G-D-MEAs) has been implemented with an automated analysis code (APE, Amperometric Peak Analysis) developed using Matlab R2022a software to provide easy and accurate detection of amperometric spike parameters, including the analysis of the pre-spike foot that sometimes precedes the complete fusion pore dilatation. Data have been acquired from cultured PC12 cells, either collecting events during spontaneous exocytosis or after L-DOPA incubation. Validation of the APE code was performed by comparing the acquired spike parameters with those obtained using Quanta Analysis (Igor macro) by Mosharov et al.

Published
2023
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7. Enhancing the Study of Quantal Exocytotic Events: Combining Diamond Multi-Electrode Arrays with Amperometric PEak Analysis (APE) an Automated Analysis Code

Author

Tomagra, Giulia, primary, Re, Alice, additional, Varzi, Veronica, additional, Aprà, Pietro, additional, Britel, Adam, additional, Franchino, Claudio, additional, Sturari, Sofia, additional, Amine, Nour-Hanne, additional, Westerink, Remco H. S., additional, Carabelli, Valentina, additional, and Picollo, Federico, additional

Published
2023
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9. Alpha-synuclein oligomers alter the spontaneous firing discharge of cultured midbrain neurons

Author

Tomagra, Giulia, Franchino, Claudio, Cesano, Federico, Chiarion, Giovanni, de Iure, Antonio, Carbone, Emilio, Calabresi, Paolo, Mesin, Luca, Picconi, Barbara, Marcantoni, Andrea, Carabelli, Valentina, Calabresi, Paolo (ORCID:0000-0003-0326-5509), Tomagra, Giulia, Franchino, Claudio, Cesano, Federico, Chiarion, Giovanni, de Iure, Antonio, Carbone, Emilio, Calabresi, Paolo, Mesin, Luca, Picconi, Barbara, Marcantoni, Andrea, Carabelli, Valentina, and Calabresi, Paolo (ORCID:0000-0003-0326-5509)

Abstract

The aim of this work was to monitor the effects of extracellular alpha-synuclein on the firing activity of midbrain neurons dissociated from substantia nigra TH-GFP mice embryos and cultured on microelectrode arrays (MEA). We monitored the spontaneous firing discharge of the network for 21 days after plating and the role of glutamatergic and GABAergic inputs in regulating burst generation and network synchronism. Addition of GABA(A), AMPA and NMDA antagonists did not suppress the spontaneous activity but allowed to identify three types of neurons that exhibited different modalities of firing and response to applied L-DOPA: high-rate (HR) neurons, low-rate pacemaking (LR-p), and low-rate non-pacemaking (LR-np) neurons. Most HR neurons were insensitive to L-DOPA, while the majority of LR-p neurons responded with a decrease of the firing discharge; less defined was the response of LR-np neurons. The effect of exogenous alpha-synuclein (alpha-syn) on the firing discharge of midbrain neurons was then studied by varying the exposure time (0-48 h) and the alpha-syn concentration (0.3-70 mu M), while the formation of alpha-syn oligomers was monitored by means of AFM. Independently of the applied concentration, acute exposure to alpha-syn monomers did not exert any effect on the spontaneous firing rate of HR, LR-p, and LR-np neurons. On the contrary, after 48 h exposure, the firing activity was drastically altered at late developmental stages (14 days in vitro, DIV, neurons): alpha-syn oligomers progressively reduced the spontaneous firing discharge (IC50 = 1.03 mu M), impaired burst generation and network synchronism, proportionally to the increased oligomer/monomer ratio. Different effects were found on early-stage developed neurons (9 DIV), whose firing discharge remained unaltered, regardless of the applied alpha-syn concentration and the exposure time. Our findings unravel, for the first time, the variable effects of exogenous alpha-syn at different stages of midbrain n

Published
2023

10. Corrigendum: Alpha-synuclein oligomers alter the spontaneous firing discharge of cultured midbrain neurons

Author

Tomagra, Giulia, Franchino, Claudio, Cesano, Federico, Chiarion, Giovanni, de Iure, Antonio, Carbone, Emilio, Calabresi, Paolo, Mesin, Luca, Picconi, Barbara, Marcantoni, Andrea, Carabelli, Valentina, Calabresi, Paolo (ORCID:0000-0003-0326-5509), Tomagra, Giulia, Franchino, Claudio, Cesano, Federico, Chiarion, Giovanni, de Iure, Antonio, Carbone, Emilio, Calabresi, Paolo, Mesin, Luca, Picconi, Barbara, Marcantoni, Andrea, Carabelli, Valentina, and Calabresi, Paolo (ORCID:0000-0003-0326-5509)

Abstract

[This corrects the article DOI: 10.3389/fncel.2023.1078550.]. N/A

Published
2023

17. Two firing modes and well-resolved Na + , K + , and Ca 2+ currents at the cell-microelectrode junction of spontaneously active rat chromaffin cell on MEAs

Author

Marcantoni, Andrea, Chiantia, Giuseppe, Tomagra, Giulia, Hidisoglu, Enis, Franchino, Claudio, Carabelli, Valentina, and Carbone, Emilio

Subjects
AP-clamp recording, Cell-electrode interface, Extracellular action potentials, Ion channel blockers, Na+, Ca2+, and K+ currents, Ca2+, Physiology, Physiology (medical), Clinical Biochemistry, Na+, and K+ currents
Abstract

We recorded spontaneous extracellular action potentials (eAPs) from rat chromaffin cells (CCs) at 37°C using microelectrode-arrays (MEAs) and compared them with intracellularly recorded APs (iAPs) through conventional patch-clamp recordings at 22°C. We show the existence of two distinct firing modes on MEAs: a ~ 4 Hz irregular continuous firing and a frequent intermittent firing mode where periods of high-intraburst frequency (~ 8 Hz) of ~ 7 s duration are interrupted by silent periods of ~ 12 s. eAPs occurred either as negative- or positive-going signals depending on the contact between cell and microelectrode: either predominantly controlled by junction-membrane ion channels (negative-going) or capacitive/ohmic coupling (positive-going).Negative-going eAPs were found to represent the trajectory of the Na+, Ca2+ and K+ currents passing through the cell area in tight contact with the microelectrode during an AP (point-contact junction). The inward Nav component of eAPs was blocked by TTX in a dose-dependent manner (IC50 ~ 10nM) while the outward component was almost fully attenuated by the BK channel blocker paxilline (200nM) or TEA (5mM). The SK channel blocker apamin (200nM) had no effect on eAPs. Inward Nav and Cav currents were well-resolved after block of Kv and BK channels in cells exhibiting no evident outward K+ currents. Unexpectedly, on the same type of cells, we could also resolve inward L-type currents after adding nifedipine (3µM). In conclusion, MEAs furnish a direct way to record different firing modes of rat CCs and to estimate the Na+, Ca2+ and K+ currents that sustain cell firing and spontaneous catecholamines secretion.

Published
2022
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20. Two firing modes and well-resolved Na+, K+, and Ca2+ currents at the cell-microelectrode junction of spontaneously active rat chromaffin cell on MEAs.

Author

Marcantoni, Andrea, Chiantia, Giuseppe, Tomagra, Giulia, Hidisoglu, Enis, Franchino, Claudio, Carabelli, Valentina, and Carbone, Emilio

Subjects
CHROMAFFIN cells, ACTION potentials, ION channels, RATS, NET Asset Value
Abstract

We recorded spontaneous extracellular action potentials (eAPs) from rat chromaffin cells (CCs) at 37 °C using microelectrode arrays (MEAs) and compared them with intracellularly recorded APs (iAPs) through conventional patch clamp recordings at 22 °C. We show the existence of two distinct firing modes on MEAs: a ~ 4 Hz irregular continuous firing and a frequent intermittent firing mode where periods of high-intraburst frequency (~ 8 Hz) of ~ 7 s duration are interrupted by silent periods of ~ 12 s. eAPs occurred either as negative- or positive-going signals depending on the contact between cell and microelectrode: either predominantly controlled by junction-membrane ion channels (negative-going) or capacitive/ohmic coupling (positive-going). Negative-going eAPs were found to represent the trajectory of the Na+, Ca2+, and K+ currents passing through the cell area in tight contact with the microelectrode during an AP (point-contact junction). The inward Nav component of eAPs was blocked by TTX in a dose-dependent manner (IC50 ~ 10 nM) while the outward component was strongly attenuated by the BK channel blocker paxilline (200 nM) or TEA (5 mM). The SK channel blocker apamin (200 nM) had no effect on eAPs. Inward Nav and Cav currents were well-resolved after block of Kv and BK channels or in cells showing no evident outward K+ currents. Unexpectedly, on the same type of cells, we could also resolve inward L-type currents after adding nifedipine (3 μM). In conclusion, MEAs provide a direct way to record different firing modes of rat CCs and to estimate the Na+, Ca2+, and K+ currents that sustain cell firing and spontaneous catecholamines secretion. [ABSTRACT FROM AUTHOR]

Published
2023
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24. Super-resolution from single photon emission: toward biological application

Author

Forneris, Jacopo, primary, Guarina, Laura, primary, Picollo, Federico, primary, Olivero, Paolo, primary, Genovese, Marco, primary, Moreva, Ekaterina, primary, Traina, Paolo, primary, Ditalia Tchernij, Sviatoslav, primary, Franchino, Claudio, primary, Ruo Berchera, Ivano, primary, Brida, Giorgio, primary, Degiovanni, Ivo P., primary, and Carabelli, Valentina, primary

Published
2017
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26. EXPLORING THE AMYLOID BETA (ABETA) DEPENDENT METABOLIC FINGERPRINT OF HIPPOCAMPAL NEURONS BY TWO-DIMENSIONAL COMPREHENSIVE GAS CHROMATOGRAPHY: POTENTIALS IN DRUG DISCOVERY STUDIES

Author

Bressanello, Davide, Garella, Davide, Bertinaria, Massimo, Marcantoni, Andrea, Franchino, Claudio, Miglio, Gianluca, Reichenbach, Stephen E., Bicchi, Carlo, and Cordero, Chiara Emilia Irma

Subjects
metabolomics, Comprehensive two-dimensional gas chromatography (GC × GC), Alzheimer's Disease, drug discovery
Published
2014

30. Methodologies for Detecting Quantal Exocytosis in Adrenal Chromaffin Cells Through Diamond-Based MEAs.

Author

Tomagra G, Franchino C, Carbone E, Marcantoni A, Pasquarelli A, Picollo F, and Carabelli V

Subjects
Catecholamines, Cells, Cultured, Cysteamine, Exocytosis physiology, Chromaffin Cells, Diamond
Abstract

Diamond-based multiarray sensors are suitable to detect in real-time exocytosis and action potentials from cultured, spontaneously firing chromaffin cells, primary hippocampal neurons, and midbrain dopaminergic neurons. Here, we focus on how amperometric measurements of catecholamine release are performed on micrographitic diamond multiarrays (μG-D-MEAs) with high temporal and spatial resolution by 16 electrodes simultaneously., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Published
2023
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