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Application of micro electrode arrays (MEAs) as an emerging technology for domoic acid- induced developmental neurotoxicity evaluation in primary cultures of rat cortical neurons
Stockholm University, Faculty of Science, The Wenner-Gren Institute .
System toxicology Unit.
System toxicology Unit.
System toxicology Unit.
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Due to lack of knowledge only a few industrial chemicals have been identified as developmental neurotoxicants. Current developmental neurotoxicity (DNT) guidelines (OECD TG 426 and EPA712-C-98-239) are based entirely on in vivo studies that are both time consuming and costly. Consequently, there is a high demand to develop alternative in vitro methods for initial screening to prioritise chemicals for further DNT testing. One of the most promising tools for neurotoxicity assessment is the measurement of electrical activity using micro electrode arrays (MEA) that provides a functional and neuronal specific endpoint that until now mainly has been used to detect acute neurotoxicity. Here, electrical activity measurements were evaluated to be a suitable endpoint for the detection of potential developmental neurotoxicants. Initially, primary cortical neurons grown on MEA were characterized for different cell markers (neural precursor cells, neurons and astrocytes) over time using immunocytochemistry to evaluate if the model could be suitable for DNT testing. Indeed, our results show that primary cortical neurons could be a promising in vitro model for DNT testing since some of the most critical neurodevelopment processes such as progenitor cell commitment, proliferation and differentiation of astrocytes and maturation of neurons are present. To evaluate if electrical activity could be a suitable endpoint to detect chemicals with DNT effects primary cortical neurons grown on MEA were exposed to domoic acid (DA), a potential developmental neurotoxicant for up to 4 weeks. Long term exposure to a low concentration (50 nM) of DA increased the basal spontaneous electrical activity as measured by spike and burst rates, as compared to the control cultures. Moreover, the effect induced by the GABAA receptor antagonist bicuculline was significantly lower in the DA treated cultures than in the untreated ones. Obtained data indicates that electrical activity measurements can be used as a tool to detect chemicals with DNT potential. However, more DNT chemicals as well as non-neurotoxic chemicals (negative controls) should be tested to confirm the use of electrical activity measurements for initial DNT screening purposes.

URN: urn:nbn:se:su:diva-30077OAI: diva2:241180
Available from: 2009-10-01 Created: 2009-10-01 Last updated: 2010-01-14Bibliographically approved
In thesis
1. Developmental Neurotoxicity Testing Using In vitro Approaches
Open this publication in new window or tab >>Developmental Neurotoxicity Testing Using In vitro Approaches
2009 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

There is a great concern about children’s health as the developing brain in foetuses and children is much more vulnerable to injury caused by different classes of chemicals than the adult brain. This vulnerability is partly due to the fact that the adult brain is well protected against chemicals by the blood brain barrier (BBB) and children have increased absorption rates and diminished ability to detoxify many exogenous compounds, in comparison to that of adults. Moreover, the development of the central nervous system (CNS) is a very complex process involving several different important events, e.g. proliferation, migration and differentiation of cells. These events are occurring within a strictly controlled time frame and therefore create different windows of vulnerability. Furthermore, the brain consists of numerous different cell types (neuronal, glial and endothelial cells) that have specific functions. The development of each cell type occurs within a specific time window and is therefore susceptible to environmental disturbances at different time periods.

Evidence indicates that exposure to industrial chemicals, pesticides or drugs, contributes to the increasing incidence of neurodevelopment disorders. However, due to lack of studies only a few industrial chemicals have been identified as developmental neurotoxicants so far. The current developmental neurotoxicity (DNT) guidelines (OECD TG 426 and US EPA 712-C-98-239) are based entirely on in vivo studies that are time consuming, complex, costly and not suitable for the testing of a high number of chemicals. Applying alternative approaches such as in silico, in vitro and non-mammalian models as a part of an integrated test strategy, could speed up the process of DNT evaluation and reduce and refine animal usage. Both in vitro and non-mammalian test systems offer the possibility of providing an early screening for a large number of chemicals, and could be particularly useful in characterising the compound-induced mechanism of toxicity of various developmental processes.

This thesis has characterised two primary neuronal cultures (cerebellar granule cells (CGCs) and cortical neuronal cultures) and identified them as relevant models for DNT testing, since the key processes of brain development are present, such as cell proliferation, migration and neuronal/glial differentiation. Furthermore, two emerging technologies (gene expression and electrical activity) have been evaluated and were identified as promising tools for in vitro DNT assessment. In combination with other assays they could be included into a DNT intelligent testing strategy to speed up the process of DNT evaluation mainly by prioritising chemicals with DNT potential for further testing.

Place, publisher, year, edition, pages
Stockholm: The Wenner-Gren Institute, Stockholm University, 2009. 64 p.
National Category
Pharmacology and Toxicology
Research subject
urn:nbn:se:su:diva-30056 (URN)978-91-7155-941-8 (ISBN)
Public defence
2009-10-30, sal E306, Arrheniuslaboratorierna, Svante Arrhenius väg 20 C, Stockholm, 10:00 (English)
The work of this thesis was performed at ECVAM, European Commission, Italy. At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: In press. Paper 3: In progress. Paper 4: In progress. Available from: 2009-10-08 Created: 2009-09-30 Last updated: 2009-10-01Bibliographically approved

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