It has been recently demonstrated that the exposure of naive neuronal cells to light—at the basis of optogenetic techniques and calcium imaging measurements—may alter neuronal firing. Indeed, understanding the effect of light on nongenetically modified neurons is crucial for a correct interpretation of calcium imaging and optogenetic experiments. Here we investigated the effect of continuous visible LED light exposure (490 nm, 0.18−1.3 mW/mm2) on spontaneous activity of primary neuronal networks derived from the early postnatal mouse cortex. We demonstrated, by calcium imaging and patch clamp experiments, that illumination higher than 1.0 mW/mm2 causes an enhancement of network activity in cortical cultures. We investigated the possible origin of the phenomena by blocking the transient receptor potential vanilloid 4 (TRPV4) channel, demonstrating a complex connection between this temperature-dependent channel and the measured effect. The results presented here shed light on an exogenous artifact, potentially present in all calcium imaging experiments, that should be taken into account in the analysis of fluorescence data.

Optonongenetic enhancement of activity in primary cortical neurons / Ghirga, S.; Pagani, F.; Rosito, M.; Angelantonio, S. D.; Ruocco, G.; Leonetti, M.. - In: JOURNAL OF THE OPTICAL SOCIETY OF AMERICA. A, OPTICS, IMAGE SCIENCE, AND VISION. - ISSN 1084-7529. - 37:4(2020), pp. 643-652. [10.1364/JOSAA.385832]

Optonongenetic enhancement of activity in primary cortical neurons

Ghirga S.
Investigation
;
Pagani F.
Investigation
;
Rosito M.
Investigation
;
Angelantonio S. D.
Supervision
;
Ruocco G.
Conceptualization
;
2020

Abstract

It has been recently demonstrated that the exposure of naive neuronal cells to light—at the basis of optogenetic techniques and calcium imaging measurements—may alter neuronal firing. Indeed, understanding the effect of light on nongenetically modified neurons is crucial for a correct interpretation of calcium imaging and optogenetic experiments. Here we investigated the effect of continuous visible LED light exposure (490 nm, 0.18−1.3 mW/mm2) on spontaneous activity of primary neuronal networks derived from the early postnatal mouse cortex. We demonstrated, by calcium imaging and patch clamp experiments, that illumination higher than 1.0 mW/mm2 causes an enhancement of network activity in cortical cultures. We investigated the possible origin of the phenomena by blocking the transient receptor potential vanilloid 4 (TRPV4) channel, demonstrating a complex connection between this temperature-dependent channel and the measured effect. The results presented here shed light on an exogenous artifact, potentially present in all calcium imaging experiments, that should be taken into account in the analysis of fluorescence data.
2020
neuron; cortical; imaging; patch clamp; network
01 Pubblicazione su rivista::01a Articolo in rivista
Optonongenetic enhancement of activity in primary cortical neurons / Ghirga, S.; Pagani, F.; Rosito, M.; Angelantonio, S. D.; Ruocco, G.; Leonetti, M.. - In: JOURNAL OF THE OPTICAL SOCIETY OF AMERICA. A, OPTICS, IMAGE SCIENCE, AND VISION. - ISSN 1084-7529. - 37:4(2020), pp. 643-652. [10.1364/JOSAA.385832]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1390202
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