Circadian clock regulates granulosa cell autophagy through NR1D1-mediated inhibition of ATG5

Zhang, Jing; Zhao, Lijia; Li, Yating; Dong, Hao; Zhang, Haisen; Ma, Tiantian; Yang, Luda; Gao, Dengke; Wang, Xiaoyu; Jiang, Haizhen; Li, Chao; Wang, Aihua; Jin, Yaping

doi:10.1152/ajpcell.00267.2021

Cited by 9 publications

(8 citation statements)

References 70 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…NR1D1, an important circadian clock component, typically acts as a transcriptional repressor of its target genes by recruiting the endogenous nuclear receptor corepressor/histone deacetylase 3 complex and plays a key role in the regulation of numerous physiological functions including lipid metabolism, immunity, and reproduction [47, 66, 67]. Our previous studies have identified an essential role for NR1D1 in steroidogenesis [31, 33, 48].…”

Section: Discussionmentioning

confidence: 99%

“…GTCs were seeded into 6-well plates (for qPCR) and 60 mm plates (for WB) for 24 h until they reached conf luence, and they were then treated with SR9009 and SR8278, respectively. For SR9009 (an agonist of NR1D1) treatment of GTCs, a concentration of 10 µ M was selected based on two previous reports [47, 48]. The non-synchronized cells were divided into two groups ( n = 3 dishes for each group) that included the SR9009 group (10 µ M, diluted in DMSO) and the control group (same volume DMSO).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Hypoxia activation attenuates progesterone synthesis in goat trophoblast cells via NR1D1 inhibition of StAR expression

Yang

et al. 2023

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

Trophoblast plays a crucial role in gestation maintenance and embryo implantation, partly due to the synthesis of progesterone. It has been demonstrated that hypoxia regulates invasion, proliferation, and differentiation of trophoblast cells. Additionally, human trophoblasts display rhythmic expression of circadian clock genes. However, it remains unclear if the circadian clock system is present in goat trophoblast cells (GTCs), and its involvement in hypoxia regulation of steroid hormone synthesis remains elusive. In this study, immunofluorescence staining revealed that both BMAL1 and NR1D1 (two circadian clock components) were highly expressed in GTCs. Quantitative real-time PCR (qPCR) analysis showed that several circadian clock genes were rhythmically expressed in forskolin-synchronized GTCs. To mimic hypoxia, GTCs were treated with hypoxia-inducing reagents (CoCl2 or DMOG). qPCR results demonstrated that hypoxia perturbed the mRNA expression of circadian clock genes and StAR. Notably, the increased expression of NR1D1 and the reduction of StAR expression in hypoxic GTCs were also detected by western blotting. In addition, progesterone secretion exhibited a notable decline in hypoxic GTCs. SR9009, an NR1D1 agonist, significantly decreased StAR expression at both the mRNA and protein levels and markedly inhibited progesterone secretion in GTCs. Moreover, SR8278, an NR1D1 antagonist, partially reversed the inhibitory effect of CoCl2 on mRNA and protein expression levels of StAR and progesterone synthesis in GTCs. Our results demonstrate that hypoxia reduces StAR expression via the activation of NR1D1 signaling in GTCs, thus inhibiting progesterone synthesis. These findings provide new insights into the NR1D1 regulation of progesterone synthesis in GTCs under hypoxic conditions.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Hypoxia activation attenuates progesterone synthesis in goat trophoblast cells via NR1D1 inhibition of StAR expression

Yang

et al. 2023

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

show abstract

“…Using other cell lines, such as hepatic stellate cells, the SR9009 did not change the autophagy markers ( 43 ). In mice granulosa cells, SR9009 upregulated Atg5 levels ( 44 ). The different results verified for the autophagic genes may be due to the dose of administration of SR9009, in addition to the different samples investigated.…”

Section: Discussionmentioning

confidence: 99%

IL-6 deletion decreased REV-ERBα protein and influenced autophagy and mitochondrial markers in the skeletal muscle after acute exercise

et al. 2022

View full text Add to dashboard Cite

Interleukin 6 (IL-6) acts as a pro and anti-inflammatory cytokine, has an intense correlation with exercise intensity, and activates various pathways such as autophagy and mitochondrial unfolded protein response. Also, IL-6 is interconnected to circadian clock-related inflammation and can be suppressed by the nuclear receptor subfamily 1, group D, member 1 (Nr1d1, protein product REV-ERBα). Since IL-6 is linked to physical exercise-modulated metabolic pathways such as autophagy and mitochondrial metabolism, we investigated the relationship of IL-6 with REV-ERBα in the adaptations of these molecular pathways in response to acute intense physical exercise in skeletal muscle. The present study was divided into three experiments. In the first one, wild-type (WT) and IL-6 knockout (IL-6 KO) mice were divided into three groups: Basal time (Basal; sacrificed before the acute exercise), 1 hour (1hr post-Ex; sacrificed 1 hour after the acute exercise), and 3 hours (3hr post-Ex; sacrificed 3 hours after the acute exercise). In the second experiment, C2C12 cells received IL-6 physiological concentrations or REV-ERBα agonist, SR9009. In the last experiment, WT mice received SR9009 injections. After the protocols, the gastrocnemius muscle or the cells were collected for reverse transcription-quantitative polymerase chain reaction (RTq-PCR) and immunoblotting techniques. In summary, the downregulation of REV-ERBα, autophagic flux, and most mitochondrial genes was verified in the IL-6 KO mice independent of exercise. The WT and IL-6 KO treated with SR9009 showed an upregulation of autophagic genes. C2C12 cells receiving IL-6 did not modulate the Nr1d1 mRNA levels but upregulated the expression of some mitochondrial genes. However, when treated with SR9009, IL-6 and mitochondrial gene expression were upregulated in C2C12 cells. The autophagic flux in C2C12 suggest the participation of REV-ERBα protein in the IL-6-induced autophagy. In conclusion, the present study verified that the adaptations required through physical exercise (increases in mitochondrial content and improvement of autophagy machinery) might be intermediated by an interaction between IL-6 and REVERBα.

show abstract

“…Exogenous addition of oxidizing LDL can trigger autophagy in human granulosa cells ( Duerrschmidt et al, 2006 ). Furthermore, the circadian clock system is also involved in the granulosa cell autophagy through the regulation of nuclear receptor subfamily 1 group D member 1 (NR1D1) ( Zhang et al, 2022 ). These findings indicate that autophagy is directly involved in the regulation of granulosa death during follicular development, whereas oxidative stress may play an important role in the regulation of granulosa autophagy.…”

Section: Discussionmentioning

confidence: 99%

Aquaporin-8 transports hydrogen peroxide to regulate granulosa cell autophagy

Huang

Jin

Zhang

et al. 2022

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Aquaporin-8 (AQP8), a member of the aquaporin family, is strongly expressed in follicular granulosa cells, which could affect the hormone secretion level in females. AQP8, as a membrane protein, could mediate H2O2 into cells, thereby triggering various biological events. The deficiency of Aqp8 increases female fertility, resulting from the decrease in follicular atresia. The low cell death rate is related to the apoptosis of granulosa cells. However, the mechanism by which AQP8 regulates the autophagy of granulosa cells remains unclear. Thus, this study aimed to explore the effect of AQP8 on autophagy in follicular atresia. We found that the expression of the autophagy marker light-chain protein 3 was significantly downregulated in the granulosa cells of Aqp8-knockout (Aqp8−/−) mice, compared with wild-type (Aqp8+/+) mice. Immunofluorescence staining and transmission electron microscopic examination indicated that the number of autophagosomes in the granulosa cells of Aqp8−/− mice decreased. Using a follicular granulosa cell autophagy model, namely a follicular atresia model, we verified that the concentration of H2O2 significantly increased during the autophagy of granulosa cells, consistent with the Aqp8 mRNA level. Intracellular H2O2 accumulation was modulated by endogenous AQP8 expression level, indicating that AQP8-mediated H2O2 was involved in the autophagy of granulosa cells. AQP8 deficiency impaired the elevation of H2O2 concentration through phosphorylated tyrosine activation. In addition, we carried out the analysis of transcriptome sequencing datasets in the ovary and found there were obvious differences in principal components, differentially expressed genes (DEGs) and KEGG pathways, which might be involved in AQP8-regulated follicular atresia. Taken together, these findings indicated that AQP8-mediated H2O2 transport could mediate the autophagy of granulosa cells. AQP8 might be a potential target for diseases related to ovarian insufficiency.

show abstract

Circadian clock regulates granulosa cell autophagy through NR1D1-mediated inhibition of ATG5

Cited by 9 publications

References 70 publications

Hypoxia activation attenuates progesterone synthesis in goat trophoblast cells via NR1D1 inhibition of StAR expression

Hypoxia activation attenuates progesterone synthesis in goat trophoblast cells via NR1D1 inhibition of StAR expression

IL-6 deletion decreased REV-ERBα protein and influenced autophagy and mitochondrial markers in the skeletal muscle after acute exercise

Aquaporin-8 transports hydrogen peroxide to regulate granulosa cell autophagy

Contact Info

Product

Resources

About