학술논문
Assessment of neurotransmitter release in human iPSC-derived neuronal/glial cells: a missing in vitro assay for regulatory developmental neurotoxicity testing.
Document Type
Academic Journal
Author
Cervetto C; Department of Pharmacy (DIFAR), Section of Pharmacology and Toxicology, University of Genoa, Italy; Interuniversity Center for the Promotion of the 3Rs Principles in Teaching and Research, Centro 3R, Pisa, Italy. Electronic address: Chiara.Cervetto@unige.it.; Pistollato F; European Commission, Joint Research Centre, JRC, Ispra, Italy.; Amato S; Department of Pharmacy (DIFAR), Section of Pharmacology and Toxicology, University of Genoa, Italy.; Mendoza-de Gyves E; European Commission, Joint Research Centre, JRC, Ispra, Italy.; Bal-Price A; European Commission, Joint Research Centre, JRC, Ispra, Italy. Electronic address: Anna.PRICE@ec.europa.eu.; Maura G; Department of Pharmacy (DIFAR), Section of Pharmacology and Toxicology, University of Genoa, Italy.; Marcoli M; Department of Pharmacy (DIFAR), Section of Pharmacology and Toxicology, University of Genoa, Italy; Interuniversity Center for the Promotion of the 3Rs Principles in Teaching and Research, Centro 3R, Pisa, Italy. Electronic address: Manuela.Marcoli@unige.it.
Source
Publisher: Pergamon In Cooperation With The Reproductive Toxicology Center Country of Publication: United States NLM ID: 8803591 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-1708 (Electronic) Linking ISSN: 08906238 NLM ISO Abbreviation: Reprod Toxicol Subsets: MEDLINE
Subject
Language
English
Abstract
Human induced pluripotent stem cell (hiPSC)-derived neural stem cells (NSCs) and their differentiated neuronal/glial derivatives have been recently considered suitable to assess in vitro developmental neurotoxicity (DNT) triggered by exposure to environmental chemicals. The use of human-relevant test systems combined with in vitro assays specific for different neurodevelopmental events, enables a mechanistic understanding of the possible impact of environmental chemicals on the developing brain, avoiding extrapolation uncertainties associated with in vivo studies. Currently proposed in vitro battery for regulatory DNT testing accounts for several assays suitable to study key neurodevelopmental processes, including NSC proliferation and apoptosis, differentiation into neurons and glia, neuronal migration, synaptogenesis, and neuronal network formation. However, assays suitable to measure interference of compounds with neurotransmitter release or clearance are at present not included, which represents a clear gap of the biological applicability domain of such a testing battery. Here we applied a HPLC-based methodology to measure the release of neurotransmitters in a previously characterized hiPSC-derived NSC model undergoing differentiation towards neurons and glia. Glutamate release was assessed in control cultures and upon depolarization, as well as in cultures repeatedly exposed to some known neurotoxicants (BDE47 and lead) and chemical mixtures. Obtained data indicate that these cells have the ability to release glutamate in a vesicular manner, and that both glutamate clearance and vesicular release concur in the maintenance of extracellular glutamate levels. In conclusion, analysis of neurotransmitter release is a sensitive readout that should be included in the envisioned battery of in vitro assays for DNT testing.
Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interests.
(Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interests.
(Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)