Acrylamide alters CREB and retinoic acid signalling pathways during differentiation of the human neuroblastoma SH-SY5Y cell line

Kristina Attoff; Ylva Johansson; Andrea Cediel-Ulloa; Jessica Lundqvist; Rajinder Gupta; Florian Caiment; Anda Gliga; Anna Forsby

doi:10.1038/s41598-020-73698-6

Acrylamide alters CREB and retinoic acid signalling pathways during differentiation of the human neuroblastoma SH-SY5Y cell line

Kristina Attoff, Ylva Johansson, Andrea Cediel-Ulloa, Jessica Lundqvist, Rajinder Gupta, Florian Caiment, Anda Gliga, Anna Forsby^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Acrylamide (ACR) is a known neurotoxicant which crosses the blood-brain barrier, passes the placenta and has been detected in breast milk. Hence, early-life exposure to ACR could lead to developmental neurotoxicity. The aim of this study was to elucidate if non-cytotoxic concentrations of ACR alter neuronal differentiation by studying gene expression of markers significant for neurodevelopment in the human neuroblastoma SH-SY5Y cell model. Firstly, by using RNASeq we identified two relevant pathways that are activated during 9 days of retinoic acid (RA) induced differentiation i.e. RA receptor (RAR) activation and the cAMP response element-binding protein (CREB) signalling pathways. Next, by qPCR we showed that 1 and 70 mu M ACR after 9 days exposure alter the expression of 13 out of 36 genes in the RAR activation pathway and 18 out of 47 in the CREB signalling pathway. Furthermore, the expression of established neuronal markers i.e. BDNF, STXBP2, STX3, TGFB1 and CHAT were down-regulated. Decreased protein expression of BDNF and altered ratio of phosphorylated CREB to total CREB were confirmed by western blot. Our results reveal that micromolar concentrations of ACR sustain proliferation, decrease neurite outgrowth and interfere with signalling pathways involved in neuronal differentiation in the SH-SY5Y cell model.

Original language	English
Article number	16714
Number of pages	15
Journal	Scientific Reports
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41598-020-73698-6
Publication status	Published - 7 Oct 2020

Keywords

FAMILY TRANSCRIPTION FACTOR
DEVELOPMENTAL NEUROTOXICITY
PROTEOMIC ANALYSIS
BINDING PROTEIN
EXPRESSION
INCREASES
EXPOSURE
GENES
BIOSYNTHESIS
METABOLISM

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@article{02573f7764714cff995f5d7bcec29457,

title = "Acrylamide alters CREB and retinoic acid signalling pathways during differentiation of the human neuroblastoma SH-SY5Y cell line",

abstract = "Acrylamide (ACR) is a known neurotoxicant which crosses the blood-brain barrier, passes the placenta and has been detected in breast milk. Hence, early-life exposure to ACR could lead to developmental neurotoxicity. The aim of this study was to elucidate if non-cytotoxic concentrations of ACR alter neuronal differentiation by studying gene expression of markers significant for neurodevelopment in the human neuroblastoma SH-SY5Y cell model. Firstly, by using RNASeq we identified two relevant pathways that are activated during 9 days of retinoic acid (RA) induced differentiation i.e. RA receptor (RAR) activation and the cAMP response element-binding protein (CREB) signalling pathways. Next, by qPCR we showed that 1 and 70 mu M ACR after 9 days exposure alter the expression of 13 out of 36 genes in the RAR activation pathway and 18 out of 47 in the CREB signalling pathway. Furthermore, the expression of established neuronal markers i.e. BDNF, STXBP2, STX3, TGFB1 and CHAT were down-regulated. Decreased protein expression of BDNF and altered ratio of phosphorylated CREB to total CREB were confirmed by western blot. Our results reveal that micromolar concentrations of ACR sustain proliferation, decrease neurite outgrowth and interfere with signalling pathways involved in neuronal differentiation in the SH-SY5Y cell model.",

keywords = "FAMILY TRANSCRIPTION FACTOR, DEVELOPMENTAL NEUROTOXICITY, PROTEOMIC ANALYSIS, BINDING PROTEIN, EXPRESSION, INCREASES, EXPOSURE, GENES, BIOSYNTHESIS, METABOLISM",

author = "Kristina Attoff and Ylva Johansson and Andrea Cediel-Ulloa and Jessica Lundqvist and Rajinder Gupta and Florian Caiment and Anda Gliga and Anna Forsby",

note = "Funding Information: The study has received funding from the Swedish Fund for Research Without Animal Experiments (N11/15 and N2018-0004), the Swedish Research Council (2018-03269) and from the European Union{\textquoteright}s Horizon 2020 research and innovation programme under the grant agreement No 681002 (EU-ToxRisk). Publisher Copyright: {\textcopyright} 2020, The Author(s).",

year = "2020",

month = oct,

day = "7",

doi = "10.1038/s41598-020-73698-6",

language = "English",

volume = "10",

journal = "Scientific Reports",

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TY - JOUR

T1 - Acrylamide alters CREB and retinoic acid signalling pathways during differentiation of the human neuroblastoma SH-SY5Y cell line

AU - Attoff, Kristina

AU - Johansson, Ylva

AU - Cediel-Ulloa, Andrea

AU - Lundqvist, Jessica

AU - Gupta, Rajinder

AU - Caiment, Florian

AU - Gliga, Anda

AU - Forsby, Anna

N1 - Funding Information: The study has received funding from the Swedish Fund for Research Without Animal Experiments (N11/15 and N2018-0004), the Swedish Research Council (2018-03269) and from the European Union’s Horizon 2020 research and innovation programme under the grant agreement No 681002 (EU-ToxRisk). Publisher Copyright: © 2020, The Author(s).

PY - 2020/10/7

Y1 - 2020/10/7

N2 - Acrylamide (ACR) is a known neurotoxicant which crosses the blood-brain barrier, passes the placenta and has been detected in breast milk. Hence, early-life exposure to ACR could lead to developmental neurotoxicity. The aim of this study was to elucidate if non-cytotoxic concentrations of ACR alter neuronal differentiation by studying gene expression of markers significant for neurodevelopment in the human neuroblastoma SH-SY5Y cell model. Firstly, by using RNASeq we identified two relevant pathways that are activated during 9 days of retinoic acid (RA) induced differentiation i.e. RA receptor (RAR) activation and the cAMP response element-binding protein (CREB) signalling pathways. Next, by qPCR we showed that 1 and 70 mu M ACR after 9 days exposure alter the expression of 13 out of 36 genes in the RAR activation pathway and 18 out of 47 in the CREB signalling pathway. Furthermore, the expression of established neuronal markers i.e. BDNF, STXBP2, STX3, TGFB1 and CHAT were down-regulated. Decreased protein expression of BDNF and altered ratio of phosphorylated CREB to total CREB were confirmed by western blot. Our results reveal that micromolar concentrations of ACR sustain proliferation, decrease neurite outgrowth and interfere with signalling pathways involved in neuronal differentiation in the SH-SY5Y cell model.

AB - Acrylamide (ACR) is a known neurotoxicant which crosses the blood-brain barrier, passes the placenta and has been detected in breast milk. Hence, early-life exposure to ACR could lead to developmental neurotoxicity. The aim of this study was to elucidate if non-cytotoxic concentrations of ACR alter neuronal differentiation by studying gene expression of markers significant for neurodevelopment in the human neuroblastoma SH-SY5Y cell model. Firstly, by using RNASeq we identified two relevant pathways that are activated during 9 days of retinoic acid (RA) induced differentiation i.e. RA receptor (RAR) activation and the cAMP response element-binding protein (CREB) signalling pathways. Next, by qPCR we showed that 1 and 70 mu M ACR after 9 days exposure alter the expression of 13 out of 36 genes in the RAR activation pathway and 18 out of 47 in the CREB signalling pathway. Furthermore, the expression of established neuronal markers i.e. BDNF, STXBP2, STX3, TGFB1 and CHAT were down-regulated. Decreased protein expression of BDNF and altered ratio of phosphorylated CREB to total CREB were confirmed by western blot. Our results reveal that micromolar concentrations of ACR sustain proliferation, decrease neurite outgrowth and interfere with signalling pathways involved in neuronal differentiation in the SH-SY5Y cell model.

KW - FAMILY TRANSCRIPTION FACTOR

KW - DEVELOPMENTAL NEUROTOXICITY

KW - PROTEOMIC ANALYSIS

KW - BINDING PROTEIN

KW - EXPRESSION

KW - INCREASES

KW - EXPOSURE

KW - GENES

KW - BIOSYNTHESIS

KW - METABOLISM

U2 - 10.1038/s41598-020-73698-6

DO - 10.1038/s41598-020-73698-6

M3 - Article

C2 - 33028897

SN - 2045-2322

VL - 10

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 16714

ER -