Oxidative phosphorylation decline and mitochondrial dynamics disequilibrium are involved in chicken large white follicle atresia

Abstract

In domestic hens, the atresia of large white follicles (LWFs) directly affects the number of follicles that enter the hierarchical development and ovulation. Figuring out factors responsible for LWFs atresia is helpful to improve egg production of hens. At the LWF stage, yellow yolk begins to be deposited into the follicles via receptor mediated endocytosis, which requires large amounts of ATP. Mitochondrial oxidative phosphorylation (OXPHOS) is the primary source of ATP for follicular development. However, it is not clear whether the OXPHOS is changed along LWFs atresia. In this study, firstly, differences in morphological appearance, histology, cell proliferation, apoptosis, OXPHOS and mitochondrial dynamics between LWFs and atretic large white follicles (ALWFs) in hens at the peak laying stage (35W) were determined to elucidate whether OXPHOS changes in ALWFs. Then, these differences of LWFs between the peak laying hens (35W-LWFs) and the late laying hens (70W-LWFs) were detected to confirm whether OXPHOS changes during LWFs atresia. The results showed that ALWFs exhibited a wrinkled surface with several hemorrhage spots, and numerous intercellular vacuoles, as well as severe nuclear pyknosis. Compared to LWFs, a higher cell apoptosis rate and a lower proliferation rate were observed in ALWFs. In ALWFs, OXPHOS declined as manifested by reductions in ATP levels, ATP synthetase abundance, NAD⁺, NADH and NAD⁺/NADH ratio, and mRNA levels of genes associated with OXPHOS complexes I-V. Meanwhile, mitochondrial dynamics disequilibrium was detected in ALWFs as the expression levels of proteins and genes related to mitochondrial fusion (MFN1, MFN2, and OPA1) decreased, while the expression levels of proteins and genes related to mitochondrial fission (DRP1 and FIS1) increased. Further, compared to 35W-LWFs, 70W-LWFs showed a histology resembling to ALWFs, manifested as a slightly loosen structure of granulosa layers, and a lower cell proliferation rate. Moreover, both lower OXPHOS and impaired mitochondrial dynamics were detected in 70W-LWFs. In conclusion, our results indicated that OXPHOS decline and mitochondrial dynamics disequilibrium are involved in LWFs atresia in laying hens.