Neurochemical Research

, Volume 41, Issue 10, pp 2574–2584 | Cite as

Prenatal Exposure to Histone Deacetylase Inhibitors Affects Gene Expression of Autism-Related Molecules and Delays Neuronal Maturation

  • Takuya Kawanai
  • Yukio Ago
  • Ryo Watanabe
  • Aya Inoue
  • Atsuki Taruta
  • Yusuke Onaka
  • Shigeru Hasebe
  • Hitoshi Hashimoto
  • Toshio Matsuda
  • Kazuhiro TakumaEmail author
Original Paper


Valproic acid (VPA) is a multi-target drug and an inhibitor of histone deacetylase (HDAC). We have previously demonstrated that prenatal exposure to VPA at embryonic day 12.5 (E12.5), but not at E14.5, causes autism-like behavioral abnormalities in male mouse offspring. We have also found that prenatal VPA exposure causes transient histone hyperacetylation in the embryonic brain, followed by decreased neuronal cell numbers in the prefrontal and somatosensory cortices after birth. In the present study, we examined whether prenatal HDAC inhibition affects neuronal maturation in primary mouse cortical neurons. Pregnant mice were injected intraperitoneally with VPA (500 mg/kg) and the more selective HDAC inhibitor trichostatin A (TSA; 500 µg/kg) at E12.5 or E14.5, and primary neuronal cultures were prepared from the cerebral cortices of their embryos. Prenatal exposure to VPA at E12.5, but not at E14.5, decreased total number, total length, and complexity of neuronal dendrites at 14 days in vitro (DIV). The effects of VPA weakened at 21 DIV. Exposure to TSA at E12.5, but not at E14.5, also delayed maturation of cortical neurons. In addition, real-time quantitative PCR revealed that the prenatal exposure to TSA decreased neuroligin-1 (Nlgn1), Shank2, and Shank3 mRNA levels and increased contactin-associated protein-like 2 mRNA level. The delay in neuronal maturation was also observed in Nlgn1-knockdown cells, which were transfected with Nlgn1 siRNA. These findings suggest that prenatal HDAC inhibition causes changes in gene expression of autism-related molecules linked to a delay of neuronal maturation.


Valproic acid Trichostatin A HDAC inhibitor Neuroligin-1 Neuronal dendrites Neuronal maturation 



Autism spectrum disorders


Bovine serum albumin

CNTNAP2 (Cntnap2)

Contactin-associated protein-like 2


Days in vitro


Histone deacetylase

NLGN (Nlgn)




NRXN1 (Nrxn1)



Phosphate-buffered saline

PTEN (Pten)

Phosphatase and tensin homolog deleted on chromosome 10


Trichostatin A


Valproic acid



This study was supported in part by KAKENHI (25460099 (YA), 26293020 (HH), 26670122 (HH), 15 H01288 (HH) and 15 K18874 (YO)), the Neuropsychiatry Drug Discovery Consortium established by Dainippon Sumitomo Pharma Co., Ltd. (Japan) with Osaka University (TM, HH), Takeda Science Foundation (Japan) (YA), Research Foundation for Pharmaceutical Sciences (Japan) (YA) and the Program for Advancing Strategic International Networks to Accelerate the Circulation of Talented Researchers (HH).

Compliance with Ethical Standards

Conflict of Interest

The authors declare they have no conflict of interest.

Supplementary material

11064_2016_1969_MOESM1_ESM.pdf (68 kb)
Supplementary material 1 (PDF 68 KB)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Takuya Kawanai
    • 1
    • 2
  • Yukio Ago
    • 1
    • 2
  • Ryo Watanabe
    • 1
  • Aya Inoue
    • 1
    • 2
  • Atsuki Taruta
    • 1
    • 2
  • Yusuke Onaka
    • 1
    • 2
  • Shigeru Hasebe
    • 1
    • 3
  • Hitoshi Hashimoto
    • 2
    • 4
  • Toshio Matsuda
    • 1
  • Kazuhiro Takuma
    • 1
    • 3
    • 4
    Email author
  1. 1.Laboratory of Medicinal Pharmacology, Graduate School of Pharmaceutical SciencesOsaka UniversitySuitaJapan
  2. 2.Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical SciencesOsaka UniversitySuitaJapan
  3. 3.Department of Pharmacology, Graduate School of DentistryOsaka UniversitySuitaJapan
  4. 4.United Graduate School of Child DevelopmentOsaka University, Kanazawa University, Hamamatsu University School of Medicine, Chiba University and University of FukuiSuitaJapan

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