Hepatocyte nuclear factor 4α (HNF4A) is a transcription factor that regulates the expression of genes in the liver, pancreas, kidney, intestine, and other tissues. Previous studies in the Mexican population have shown a high frequency of the Thr130Ile polymorphism and have suggested its important role in the pathogenesis of early-onset type 2 diabetes. The aim of the present study was to determine whether this variant also contributes to gestational diabetes mellitus (GDM) in a Mexican population. We studied 213 unrelated postpartum women and their neonates, who were divided into 2 groups: control and GDM. The control group was formed by 108 healthy postpartum women and their neonates, and the GDM group was formed by 105 postpartum women diagnosed with GDM and their neonates. All subjects were genotyped for the Thr130Ile polymorphism in HNF4A by Taqman allelic discrimination assays and sequencing. Our results showed a higher frequency of the minor allele of the Thr130Ile polymorphism in the GDM group compared with the control group (P = 0.0452; odds ratio, 2.59; 95% confidence interval, 1.02-6.59). With respect to offspring, the frequency of the polymorphism was higher in the offspring of the GDM group than in the offspring of the control group; however, no significant differences between the groups were observed (P = 0.2551; odds ratio, 1.90; 95% confidence interval, 0.99-3.64). The findings suggest that the Thr130Ile polymorphism is associated with GDM in the studied Mexican population.