title: Acrylamide alters neurotransmitter induced calcium responses in murine ESC-derived and primary neurons.
abstract: Stem cell-derived specialized cell types are of interest as an alternative cell system to identify and research neurotoxic effects and modes of action. Developmental toxicity may be studied during differentiation, while organ-specific toxicity may be assessed in fully functional cells, such as neurons. In this study we tested if fully differentiated neurons derived from murine embryonic stem cells (ESCN) could be used to investigate the effects of the well characterized neurotoxic model compound acrylamide (ACR) and if ESCN behave similar to murine primary cortical neurons (pCN) from 16 days old embryos. We characterized the differentiation process of cryopreserved ESC-derived neural precursor cells (NPC) differentiating to ESCN. During the differentiation process (days 11-20) a strong increase in calcium responses to glutamate, acetylcholine and GABA were observed. Moreover, neuron specific marker proteins, β-III-tubulin, MAP2, Tau, Rbfox3 and synaptophysin showed similar patterns to pCN. In ESCN and pCN the neuronal structure, e.g. neurites, was not affected by low concentrations of ACR [0.5-1.6mM]. However, 24h incubation periods with 0.5-1.6mM ACR led to a reduction of acetylcholine and glutamate induced calcium responses. In conclusion, we show that non-cytotoxic concentrations of ACR alter neurotransmission in ESCN as well as pCN.
authors.affiliation: IfADo - Leibniz Research Center for Working Environment and Human Factors, Dortmund, Germany. Electronic address: julia.sisnaiske@rub.de.
authors.forename: Julia
authors.initials: J
authors.lastname: Sisnaiske
authors.affiliation: IfADo - Leibniz Research Center for Working Environment and Human Factors, Dortmund, Germany.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Hausherr
authors.affiliation: University of Konstanz, Doerenkamp-Zbinden Chair for In Vitro Toxicology and Biomedicine, Konstanz, Germany.
authors.forename: Anne K
authors.initials: AK
authors.lastname: Krug
authors.affiliation: University of Konstanz, Doerenkamp-Zbinden Chair for In Vitro Toxicology and Biomedicine, Konstanz, Germany; Center for Stem Cell Biology, Sloan-Kettering Institute for Cancer Research, New York City, USA; Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, New York City, USA.
authors.forename: Bastian
authors.initials: B
authors.lastname: Zimmer
authors.affiliation: IfADo - Leibniz Research Center for Working Environment and Human Factors, Dortmund, Germany.
authors.forename: Jan G
authors.initials: JG
authors.lastname: Hengstler
authors.affiliation: University of Konstanz, Doerenkamp-Zbinden Chair for In Vitro Toxicology and Biomedicine, Konstanz, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Leist
authors.affiliation: IfADo - Leibniz Research Center for Working Environment and Human Factors, Dortmund, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: van Thriel
mesh_terms: D020106:Acrylamide / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism; D016501:Neurites / Q000187:drug effects; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism*
chemical_list: D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; D020106:Acrylamide; D002118:Calcium
keywords: 3R; Acrylamide; Neuronal function; Neurotoxicity testing; Stem cell