Prenatal 4-phenylbutyric acid administration during mid-gestation ameliorates ASD-like behaviors by reducing cortical endoplasmic reticulum stress in VPA-induced ICR and BTBR mouse models

Koichi Kawada; Seisuke Mimori; Nobuyuki Kuramoto; Kyosuke Uno; Koichi Kawada; Seisuke Mimori; Nobuyuki Kuramoto; Kyosuke Uno

doi:10.3934/Neuroscience.2026002

AIMS Neuroscience

2026, Volume 13, Issue 1: 29-49. doi: 10.3934/Neuroscience.2026002

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Research article

Prenatal 4-phenylbutyric acid administration during mid-gestation ameliorates ASD-like behaviors by reducing cortical endoplasmic reticulum stress in VPA-induced ICR and BTBR mouse models

1.
Laboratory of Molecular Pharmacology, Faculty of Pharmaceutical Sciences, Setsunan University, 45–1 Nagaotoge-cho, Hirakata, Osaka 573–0101, Japan
2.
Department of Pharmacology, Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15–8 Shiomi-cho, Choshi, Chiba 288–0025, Japan
3.
Department of Clinical Medicine, Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15–8 Shiomi-cho, Choshi, Chiba 288–0025, Japan

Received: 04 November 2025 Revised: 29 December 2025 Accepted: 06 January 2026 Published: 20 January 2026

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by deficits in social interaction and repetitive behaviors. Increasing evidence suggests that endoplasmic reticulum (ER) stress contributes to abnormal brain development in ASD; however, whether prenatal modulation of ER stress can prevent ASD-like phenotypes remains unclear. In this study, we investigated the effects of prenatal administration of the chemical chaperone 4-phenylbutyric acid (4-PBA) in two etiologically distinct ASD mouse models: valproic acid (VPA)-exposed Jcl:ICR (ICR) mice and BTBR T⁺ Itpr3^tf/J (BTBR) mice. Social behaviors were evaluated using the three-chamber test, and repetitive behaviors were assessed by self-grooming duration. 4-PBA was administered to mid-gestation mice, and behavioral changes in the male offspring derived-two type ASD model (VPA and BTBR mice) were evaluated. 4-PBA reduced ER stress in the cerebral cortex of the offspring male VPA and BTBR mice. In particular, 4-PBA strongly inhibited the expression of 94-kDa glucose-regulated protein, an ER stress marker, in BTBR male mice. In addition, 4-PBA improved synaptic organizer expression and neuronal maturation, which are diminished in ASD, specifically in the cerebral cortex of VPA mice. Furthermore, 4-PBA improved social reciprocity, a behavior specific to ASD in male VPA and BTBR mice. In conclusion, ER stress during mid-pregnancy is strongly associated with the development of ASD symptoms. Furthermore, the reduction in ER stress by 4-PBA leads to the suppression of ASD symptoms. Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by a lack of sociality, and the difficulty in forming social relationships often leads to various social problems. Prenatal administration of 4-PBA to mothers may reduce the risk of developing ASD, and we believe this could be a new approach to prevention.
- autism spectrum disorder,
- sociability,
- endoplasmic reticulum stress,
- glucose-regulated protein,
- 4-phenylbutyric acid
Citation: Koichi Kawada, Seisuke Mimori, Nobuyuki Kuramoto, Kyosuke Uno. Prenatal 4-phenylbutyric acid administration during mid-gestation ameliorates ASD-like behaviors by reducing cortical endoplasmic reticulum stress in VPA-induced ICR and BTBR mouse models[J]. AIMS Neuroscience, 2026, 13(1): 29-49. doi: 10.3934/Neuroscience.2026002

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Abstract

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by deficits in social interaction and repetitive behaviors. Increasing evidence suggests that endoplasmic reticulum (ER) stress contributes to abnormal brain development in ASD; however, whether prenatal modulation of ER stress can prevent ASD-like phenotypes remains unclear. In this study, we investigated the effects of prenatal administration of the chemical chaperone 4-phenylbutyric acid (4-PBA) in two etiologically distinct ASD mouse models: valproic acid (VPA)-exposed Jcl:ICR (ICR) mice and BTBR T⁺ Itpr3^tf/J (BTBR) mice. Social behaviors were evaluated using the three-chamber test, and repetitive behaviors were assessed by self-grooming duration. 4-PBA was administered to mid-gestation mice, and behavioral changes in the male offspring derived-two type ASD model (VPA and BTBR mice) were evaluated. 4-PBA reduced ER stress in the cerebral cortex of the offspring male VPA and BTBR mice. In particular, 4-PBA strongly inhibited the expression of 94-kDa glucose-regulated protein, an ER stress marker, in BTBR male mice. In addition, 4-PBA improved synaptic organizer expression and neuronal maturation, which are diminished in ASD, specifically in the cerebral cortex of VPA mice. Furthermore, 4-PBA improved social reciprocity, a behavior specific to ASD in male VPA and BTBR mice. In conclusion, ER stress during mid-pregnancy is strongly associated with the development of ASD symptoms. Furthermore, the reduction in ER stress by 4-PBA leads to the suppression of ASD symptoms. Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by a lack of sociality, and the difficulty in forming social relationships often leads to various social problems. Prenatal administration of 4-PBA to mothers may reduce the risk of developing ASD, and we believe this could be a new approach to prevention.

Abbreviations

ASD

autism spectrum disorder

CADM1

cell adhesion molecule 1

CNS

central nerve system

endoplasmic reticulum

ERAD

endoplasmic reticulum associated degradation

GRP78

Glucose-Regulated Protein 78-kDa

GRP94

Glucose-Regulated Protein 94-kDa

HDAC

histone deacetylase

MAP-2

microtubule-associated protein-2

NLGN

neuroligin

4-PBA

4-phenylbutyric acid

PSD-95

postsynaptic density protein-95

UPR

unfolded protein response

VPA

valproic acid

Acknowledgments

This work was supported by the Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research (grant number: JP15K18879). The authors would like to thank Editage (www.editage.jp) for assistance with English language editing.

Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Conflict of interest

The authors declare no conflict of interest.

Authors' contributions

Conceptualization: KK; Data curation: KK; Formal analysis: KK, SM; Funding acquisition: KK; Investigation: KK, SM; Methodology: KK, SM; Project administration: KK; Resources: KK; Software: N/A; Supervision: KK, NK; Validation: KK, SM, KU; Visualization: KK; Writing-original draft: KK; Writing-review and editing: KK, SM, NK, KU. Initials are Koichi Kawada (KK), Seisuke Mimori (SM), Nobuyuki Kuramoto (NK) and Kyosuke Uno (KU). All authors approved the final version for publication.

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