Satou Ryouichi, Sato Masaki, Kimura Maki, Ishizuka Yoichi, Tazaki Masakazu, Sugihara Naoki, Shibukawa Yoshiyuki
Department of Epidemiology and Public Health, Tokyo Dental CollegeTokyo, Japan.
Department of Physiology, Tokyo Dental CollegeTokyo, Japan.
Front Physiol. 2017 May 23;8:320. doi: 10.3389/fphys.2017.00320. eCollection 2017.
Circadian rhythms are essential for health and regulate various physiological functions. These rhythms are regulated by a negative-feedback loop involving clock genes in the suprachiasmatic nucleus (SCN) and peripheral tissues. The rate of secretion of salivary substances, ions, and water follows a circadian rhythm, however, the relationship between the molecular mechanism of salivary secretion and peripheral circadian rhythm is not yet clear. Anoctamin 1 (ANO1, also known as TMEM16A) and Aquaporin 5 (AQP5) play an important role in the transport of ions and water in the submandibular glands (SGs). We examined the interaction between the rhythmic expression pattern of the clock genes, and , in rat whole SGs as well as isolated acinar and ductal cells. Circadian rhythmic expression for α, and α mRNAs, also called the clock genes, was observed in rat SGs by semi-quantitative RT-PCR analysis. We also observed rhythmic patterns in and mRNA expression. The expression of ANO1 protein also showed circadian rhythm, as confirmed by western blot analysis. We could not observe any time delay between the peak expression of ANO1 protein and its mRNA. Expression levels of the clock gene mRNAs in the ductal cells was higher than that in acinar cells, however, rhythmic oscillations were observed in both. Our results suggest that SGs have peripheral clocks, and rhythmic expressions of and along with the clock genes, may play an important role in the circadian regulation of salivary secretion.
昼夜节律对健康至关重要,并调节各种生理功能。这些节律由一个负反馈回路调节,该回路涉及视交叉上核(SCN)和外周组织中的时钟基因。唾液物质、离子和水的分泌速率遵循昼夜节律,然而,唾液分泌的分子机制与外周昼夜节律之间的关系尚不清楚。anoctamin 1(ANO1,也称为TMEM16A)和水通道蛋白5(AQP5)在颌下腺(SGs)的离子和水运输中起重要作用。我们研究了时钟基因、在大鼠全SGs以及分离的腺泡和导管细胞中的节律性表达模式之间的相互作用。通过半定量RT-PCR分析在大鼠SGs中观察到了α和α mRNA(也称为时钟基因)的昼夜节律性表达。我们还观察到了和mRNA表达的节律模式。蛋白质印迹分析证实,ANO1蛋白的表达也呈现昼夜节律。我们没有观察到ANO1蛋白与其mRNA峰值表达之间的任何时间延迟。导管细胞中时钟基因mRNA的表达水平高于腺泡细胞,但两者均观察到节律性振荡。我们的结果表明,SGs具有外周时钟,并且、与时钟基因的节律性表达可能在唾液分泌的昼夜调节中起重要作用。