Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons

Abstract A finely tuned Ca2+ homeostasis in restricted cell domains is of fundamental importance for neurons, where transient Ca2+ oscillations direct the proper coordination of electro-chemical signals and overall neuronal metabolism. Once such a precise regulation is unbalanced, however, neuronal...

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Autores principales:	Rosa Pia Norante, Maria Lina Massimino, Paolo Lorenzon, Agnese De Mario, Caterina Peggion, Mattia Vicario, Mattia Albiero, Maria Catia Sorgato, Raffaele Lopreiato, Alessandro Bertoli
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8e299e8e280d44439975e49e38da4983
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spelling	oai:doaj.org-article:8e299e8e280d44439975e49e38da49832021-12-02T11:40:30ZGeneration and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons10.1038/s41598-017-06919-02045-2322https://doaj.org/article/8e299e8e280d44439975e49e38da49832017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06919-0https://doaj.org/toc/2045-2322Abstract A finely tuned Ca2+ homeostasis in restricted cell domains is of fundamental importance for neurons, where transient Ca2+ oscillations direct the proper coordination of electro-chemical signals and overall neuronal metabolism. Once such a precise regulation is unbalanced, however, neuronal functions and viability are severely compromised. Accordingly, disturbed Ca2+ metabolism has often been claimed as a major contributor to different neurodegenerative disorders, such as amyotrophic lateral sclerosis that is characterised by selective motor neuron (MN) damage. This notion highlights the need for probes for the specific and precise analysis of local Ca2+ dynamics in MNs. Here, we generated and functionally validated adeno-associated viral vectors for the expression of gene-encoded fluorescent Ca2+ indicators targeted to different cell domains, under the transcriptional control of a MN-specific promoter. We demonstrated that the probes are specifically expressed, and allow reliable local Ca2+ measurements, in MNs from murine primary spinal cord cultures, and can also be expressed in spinal cord MNs in vivo, upon systemic administration to newborn mice. Preliminary analyses using these novel vectors have shown larger cytosolic Ca2+ responses following stimulation of AMPA receptors in the cytosol of primary cultured MNs from a murine genetic model of ALS compared to the healthy counterpart.Rosa Pia NoranteMaria Lina MassiminoPaolo LorenzonAgnese De MarioCaterina PeggionMattia VicarioMattia AlbieroMaria Catia SorgatoRaffaele LopreiatoAlessandro BertoliNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rosa Pia Norante Maria Lina Massimino Paolo Lorenzon Agnese De Mario Caterina Peggion Mattia Vicario Mattia Albiero Maria Catia Sorgato Raffaele Lopreiato Alessandro Bertoli Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons
description	Abstract A finely tuned Ca2+ homeostasis in restricted cell domains is of fundamental importance for neurons, where transient Ca2+ oscillations direct the proper coordination of electro-chemical signals and overall neuronal metabolism. Once such a precise regulation is unbalanced, however, neuronal functions and viability are severely compromised. Accordingly, disturbed Ca2+ metabolism has often been claimed as a major contributor to different neurodegenerative disorders, such as amyotrophic lateral sclerosis that is characterised by selective motor neuron (MN) damage. This notion highlights the need for probes for the specific and precise analysis of local Ca2+ dynamics in MNs. Here, we generated and functionally validated adeno-associated viral vectors for the expression of gene-encoded fluorescent Ca2+ indicators targeted to different cell domains, under the transcriptional control of a MN-specific promoter. We demonstrated that the probes are specifically expressed, and allow reliable local Ca2+ measurements, in MNs from murine primary spinal cord cultures, and can also be expressed in spinal cord MNs in vivo, upon systemic administration to newborn mice. Preliminary analyses using these novel vectors have shown larger cytosolic Ca2+ responses following stimulation of AMPA receptors in the cytosol of primary cultured MNs from a murine genetic model of ALS compared to the healthy counterpart.
format	article
author	Rosa Pia Norante Maria Lina Massimino Paolo Lorenzon Agnese De Mario Caterina Peggion Mattia Vicario Mattia Albiero Maria Catia Sorgato Raffaele Lopreiato Alessandro Bertoli
author_facet	Rosa Pia Norante Maria Lina Massimino Paolo Lorenzon Agnese De Mario Caterina Peggion Mattia Vicario Mattia Albiero Maria Catia Sorgato Raffaele Lopreiato Alessandro Bertoli
author_sort	Rosa Pia Norante
title	Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons
title_short	Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons
title_full	Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons
title_fullStr	Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons
title_full_unstemmed	Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons
title_sort	generation and validation of novel adeno-associated viral vectors for the analysis of ca2+ homeostasis in motor neurons
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/8e299e8e280d44439975e49e38da4983
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Generation and validation of novel adeno-associated viral vectors for the analysis of Ca2+ homeostasis in motor neurons

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