Archives of Toxicology

, Volume 93, Issue 7, pp 1993–2006 | Cite as

Proteomic analysis of hippocampal proteins in acrylamide-exposed Wistar rats

  • Daichi Nagashima
  • Lingyi Zhang
  • Yuki Kitamura
  • Sahoko Ichihara
  • Eri Watanabe
  • Cai Zong
  • Yuko Yamano
  • Toshihiro Sakurai
  • Shinji Oikawa
  • Gaku IchiharaEmail author
Organ Toxicity and Mechnisms


Acrylamide has been used industrially and also found in certain foods cooked at high temperatures. Previous reports described acrylamide-related human intoxication who presented with ataxia, memory impairment, and/or illusion. The aim of this study was to characterize the molecular mechanisms of neurotoxicity of acrylamide by analyzing the expression levels of various proteins in the hippocampus of rats exposed to acrylamide. Male Wistar rats were administered acrylamide by gavage at 0, 2, and 20 mg/kg for 1 week or 0, 0.2, 2, and 20 mg/kg for 5 weeks. At the end of the experiment, the hippocampus was dissected out and proteins were extracted for two-dimensional difference gel electrophoresis combined with matrix-assisted laser-desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF/MS). MALDI-TOF/TOF/MS identified significant changes in two proteins in the 1-week and 22 proteins in the 5-week exposure groups. These changes were up-regulation in 9 and down-regulation in 13 proteins in the hippocampus of rats exposed to acrylamide at 20 mg/kg for 5 weeks. PANTHER overrepresentation test based on the GO of biological process showed significant overrepresentation in proteins annotated to nicotinamide nucleotide metabolic process, coenzyme biosynthetic process, pyruvate metabolic process, and carbohydrate metabolic process. The test also showed significant overrepresentation in proteins annotated to creatinine kinase activity for the GO of molecular function as well as myelin sheath, cytoplasmic part, and cell body for the GO of cellular component. Comparison with a previous proteomic study on hippocampal proteins in rats exposed to 1-bromopropane identified triosephosphate isomerase, mitochondrial creatine kinase U-type, creatine kinase β-type and proteasome subunit α type-1 as proteins affected by exposure to acrylamide and 1-bromopropane, suggesting a common mechanism of neurotoxicity for soft electrophiles.


Acrylamide Proteomic analysis Neurotoxicology Triosephosphate isomerase Soft electrophiles 



This work was supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science 26293148, 16H02965, 17H06396, and 17H07105. The authors thank Ms. Yoshiko Murakata for the generous help in proteomic analysis, Dr. Xiao Zhang for help in molecular analysis, and Ms. Yurina Wakayama, Mr. Kyo Morita, Ms. Hana Katano, and Ms. Mami Sakai for their generous help in the experiment. We thank Ms. Satoko Arai for the excellent secretarial support. We also acknowledge the help and support of Dr. Toshio Nakadate throughout the study.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

204_2019_2484_MOESM1_ESM.pptx (92 kb)
Supplementary Fig. 1 Changes in body weight of rats exposed by gavage to acrylamide at 0, 2, 20 mg/kg bw for 1 week or 0, 0.2, 2, 20 mg/kg bw for 5 weeks. Daily exposure to acrylamide at 20mg/kg bw was associated with a significant body weight loss from Day 30. The value is the mean ± SD (n=5 rats in each group), *p < 0.05, compared with the control by one-way ANOVA followed by Dunnett’s test. Supplementary Fig. 2 a, b, c, d) Changes in weight of different body organs during the 1-week study period. Numbers on the abscissa represent the dose of acrylamide (0 mg/kg, control group; 2 mg/kg group; 20 mg/kg group. n=6 in each group). e, f, g, h, I, j) Changes in weight of different body organs during the 5-week study period. Numbers on the abscissa represent the dose of acrylamide (0 mg/kg, control group, n =5; 0.2 mg/kg group, n=3; 2 mg/kg group, n=5; 20 mg/kg group, n=5). The weight of the brain, adrenal gland, liver, kidney and spleen at day 35 was significantly lower in the 20 mg/kg acrylamide group compared with the control. Data are mean ± SD, *p<0.05, compared with the control, by one-way ANOVA followed by Dunnett’s test (PPTX 91 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Daichi Nagashima
    • 1
    • 2
  • Lingyi Zhang
    • 1
  • Yuki Kitamura
    • 3
    • 5
  • Sahoko Ichihara
    • 4
  • Eri Watanabe
    • 1
  • Cai Zong
    • 1
  • Yuko Yamano
    • 2
  • Toshihiro Sakurai
    • 1
  • Shinji Oikawa
    • 3
  • Gaku Ichihara
    • 1
    Email author
  1. 1.Department of Occupational and Environmental Health, Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan
  2. 2.Department of Hygiene and Preventive MedicineShowa UniversityShinagawaJapan
  3. 3.Department of Environmental and Molecular MedicineMie University Graduate School of MedicineMieJapan
  4. 4.Department of Environmental and Preventive MedicineJichi Medical University School of MedicineShimotsukeJapan
  5. 5.College of PharmacyKinjo Gakuin UniversityAichiJapan

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