Aim:Antiepileptic drugs (AEDs) have been known to cause congenital malformations, including neural tube defects. However, inadequate data are available regarding the effects of recently developed AEDs, such as pregabalin. The present study was conducted to evaluate the effect of pregabalin on neural tube development using early chick embryos.Material and Methods:Experiments were conducted on specific pathogen-free Leghorn chick embryos, which were equally divided into control and pregabalin-treated (therapeutic dose, 600 mg; supratherapeutic dose, 1200 mg) groups. The embryos were macroscopically and microscopically evaluated following pregabalin administration. Expression levels of mammalian target of rapamycin (mTOR), c-Jun N-terminal kinase (JNK), and microtubule-associated proteins 1A/1B light chain 3 (LC3) in the embryos were observed.Results:The embryos in the therapeutic dose group appeared more curved than those in the control group. The vesicles in the supratherapeutic dose group were more distinct but smaller than those in the control and therapeutic dose groups. mTOR expression was high in the control group and low in the therapeutic and supratherapeutic dose groups. JNK expression was low in the control and therapeutic dose groups and moderate in the supratherapeutic dose group. LC3 expression was moderate in the control and therapeutic dose groups and strong in the supratherapeutic dose group.Conclusion:Pregabalin administration induced neural tube defects and fetal abnormalities in the chick embryos through increased autophagy due to enhanced apoptosis in the prenatal fetus.