Karolinska Institutet, Department of Medicine H7, Huddinge, Sweden; University of Sao Paulo, Department of Physiology and Biophysics, Sao Paulo, Brazil.
University of Sao Paulo, Department of Physiology and Biophysics, Sao Paulo, Brazil.
Life Sci. 2018 Aug 15;207:36-41. doi: 10.1016/j.lfs.2018.05.043. Epub 2018 May 26.
Daily and seasonal rhythms coordinate the endocrine and metabolic functions. The pituitary gland is the master regulator of several endocrine activities, and its function is classically regulated by endocrine signals from its target glands as well as from the hypothalamus. The growth hormone (GH) produced and secreted by the anterior pituitary presents a pulsatile secretion throughout the 24-hour cycle. However, the molecular mechanisms regulating the daily pattern of GH secretion are still unclear. Herein we investigated whether circadian GH mRNA and protein synthesis is modulated by acute adjustments in the stability and expression of GH mRNA.
GH mRNA and protein content were evaluated by real-time PCR and Western blotting, respectively, in pituitary gland of rats euthanized every 3 h during a 24-h period at the Zeitgeber times (ZT3 to ZT24). The GH mRNA poly(A) tail length was determined by RACE-PAT assay.
We identified two main peaks of GH mRNA level in the pituitary gland of rats; one in the middle of the light-cycle and another in the middle of the dark-cycle. The latter was associated with an increase in pituitary GH protein content. Interestingly, an increment in the poly(A) tail length of the GH transcript was observed in association to reduced migration rate of the GH transcript and increased mRNA content in the dark-cycle period.
Our findings provide evidence that changes in the GH mRNA poly(A) length may underlie the circadian pattern of GH mRNA and protein levels in the pituitary gland of rats.
昼夜节律协调内分泌和代谢功能。垂体是几种内分泌活动的主调控器,其功能经典地受到来自靶腺和下丘脑的内分泌信号的调节。由垂体前叶产生和分泌的生长激素(GH)在 24 小时周期内呈现脉冲式分泌。然而,调节 GH 分泌日节律的分子机制尚不清楚。在此,我们研究了急性调整 GH mRNA 的稳定性和表达是否调节昼夜 GH mRNA 和蛋白的合成。
在 Zeitgeber 时间(ZT3 至 ZT24)期间,每天处死大鼠 3 小时,分别通过实时 PCR 和 Western 印迹法评估垂体中 GH mRNA 和蛋白含量。通过 RACE-PAT 测定法确定 GH mRNA 聚(A)尾的长度。
我们在大鼠垂体中鉴定出 GH mRNA 水平的两个主要峰值;一个在光周期的中间,另一个在暗周期的中间。后者与垂体 GH 蛋白含量的增加有关。有趣的是,在暗周期期间,GH 转录物的聚(A)尾长增加,与 GH 转录物迁移率降低和 mRNA 含量增加有关。
我们的研究结果提供了证据,表明 GH mRNA 聚(A)长度的变化可能是大鼠垂体中 GH mRNA 和蛋白水平昼夜节律模式的基础。