University of Novi Sad, Faculty of Sciences, Department of Biology and Ecology, Laboratory for Chronobiology and Aging, Laboratory for Reproductive Endocrinology and Signaling, Novi Sad, Serbia.
Biofactors. 2024 Jul-Aug;50(4):738-749. doi: 10.1002/biof.2035. Epub 2023 Dec 26.
The involvement of the molecular clock in regulating cell physiological processes on a specific time scale is a recognized concept, yet its specific impact on optimizing androgen production in Leydig cells has been unclear. This study aimed to confirm the role of the REVERBA (NR1D1) gene in controlling the transcription of key genes related to Leydig cell steroid production. We investigated daily variations by collecting Leydig cells from rats at various times within a 24-h period. Chromatin immunoprecipitation study showed a time-dependent pattern for genes linked to steroid production (Nur77, Star, Cyp11a1, and Cyp17a1), which closely matched the 24-h REVERBA levels in Leydig cells, peaking between zeitgeber time (ZT) 7-11. To understand the physiological significance of REVERBA's interaction with promoters of steroidogenesis-related genes, Leydig cells from rats at two different times (ZT7 and ZT16; chosen based on REVERBA expression levels), were treated with either an agonist (GSK4112) or an antagonist (SR8278). The results revealed that the REVERBA agonist stimulated gene transcription, while the antagonist inhibited it, but only when REVERBA was sufficiently present, indicating a reliance on REVERBA's circadian fluctuation. Moreover, this REVERBA-dependent stimulation had a clear impact on testosterone production in the culture medium, underscoring REVERBA's involvement in the circadian regulation of testosterone. This study indicates that REVERBA, in addition to being a core component of the cellular clock, plays a key role in regulating androgen production in Leydig cells by influencing the transcription of critical steroidogenesis-related genes.
分子钟参与调节特定时间尺度上的细胞生理过程是一个公认的概念，但它对优化睾丸间质细胞雄激素产生的具体影响尚不清楚。本研究旨在证实 REVERBA（NR1D1）基因在控制与睾丸间质细胞类固醇产生相关的关键基因转录中的作用。我们通过在 24 小时内的不同时间从大鼠中收集睾丸间质细胞来研究日变化。染色质免疫沉淀研究显示，与类固醇产生相关的基因（Nur77、Star、Cyp11a1 和 Cyp17a1）的转录具有时间依赖性模式，与 Leydig 细胞中 24 小时 REVERBA 水平密切匹配，在 Zeitgeber 时间（ZT）7-11 之间达到峰值。为了了解 REVERBA 与类固醇生成相关基因启动子相互作用的生理意义，我们在两个不同时间（ZT7 和 ZT16；根据 REVERBA 表达水平选择）从大鼠中分离睾丸间质细胞，并用激动剂（GSK4112）或拮抗剂（SR8278）处理。结果表明，REVERBA 激动剂刺激基因转录，而拮抗剂抑制基因转录，但只有在 REVERBA 充分存在的情况下才会发生这种情况，表明依赖于 REVERBA 的昼夜节律波动。此外，这种依赖于 REVERBA 的刺激对培养物中睾酮的产生有明显影响，突出了 REVERBA 参与睾酮的昼夜节律调节。这项研究表明，REVERBA 除了作为细胞时钟的核心组成部分外，还通过影响关键类固醇生成相关基因的转录，在调节睾丸间质细胞雄激素产生中发挥关键作用。
