Journal
THERIOGENOLOGY
Volume 202, Issue -, Pages 51-60Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.theriogenology.2023.03.004
Keywords
CLOCK; Estradiol synthesis; Mitochondrial function; Granulosa cells; Pig
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The CLOCK gene plays a critical role in estradiol synthesis in porcine granulosa cells. The alteration of the CLOCK gene leads to impaired mitochondrial function and reduced estradiol synthesis.
Circadian locomotor output cycles kaput (CLOCK) is a critical component of the mammalian circadian clock system and regulates ovarian physiology. However, the functions and mechanisms of CLOCK in porcine granulosa cells (GCs) are poorly understood. The present study focused on CLOCK's effects on estradiol synthesis. Similarity analysis showed that CLOCK is highly conserved between pigs and other species. The phylogenetic tree analysis indicated that porcine CLOCK was most closely related to that in Arabian camels. CLOCK significantly reduced E2 synthesis in GCs. CLOCK reduced the expression of steroidogenesis-related genes at the mRNA and protein levels, including CYP19A1, CYP11A1, and StAR. CYP17A1 levels were significantly downregulated. We demonstrated that CLOCK dramatically decreased ATP content, mitochondrial copy number, and mitochondrial membrane potential (MMP) and increased reactive oxygen species levels in GCs. We observed that mitochondria were severely damaged with fuzzy and fractured cristae and swollen matrix. These findings suggest that mitochondrial function and E2 synthesis are impaired following the alteration of CLOCK gene expression in porcine ovarian GCs. (c) 2023 Elsevier Inc. All rights reserved.
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