Vacuolar protein sorting 36 (VPS36), a protein primarily known for its role in the Endosomal Sorting Complex Required for Transport pathway, has recently been shown to be linked to chicken reproduction. Previous research showed that is significantly downregulated in sexually mature chicken ovaries compared to immature ones. In this study, using real-time quantitative RT-PCR, we investigated the expression pattern of and its head-to-head gene mRNA in chicken follicles. Small white follicles were found to have significantly higher expression of and mRNA than any other sized follicles ( <0.05). The expression of and mRNA were detected in both granulosa and theca layers of pre-ovulatory follicles, the expression of in theca layers was slightly higher than in granulosa cells. Treatment of small yellow follicles with follicle-stimulating hormone and estradiol resulted in a marked decrease of both and mRNA ( <0.05); however, progesterone, transforming growth factor-β 1 and luteinizing hormone induced no significant changes in and mRNA expression in these follicles. These results indicate that the head-to-head genes of and exhibit similar expression in chicken follicles and are involved in chicken follicle development.