2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38797Chronic restricted sleep or sleep deprivation is a common health problem for many people in our industrial societies. Insufficiency of sleep would cause alterations in brain functions and physiological behaviors including activities of important enzymes, neurotransmitter levels, impairment of neuronal excitability and circadian rhythmicity. Previous studies have indicated that the circadian center of mammalian is located in the suprachiasmatic nucleus (SCN) of anterior hypothalamus. The SCN neurons have a near 24-hour rhythm of electrical activity that is determined by the slowly inactivating component of Na+ current. By the way of the diverse projections from SCN to numerous autonomic centers, SCN could impose its rhythmic activation to lots of biological activities ranging from hormonal secretion to sleep-wake cycle operation. It is indicated that the neuronal output signal of SCN is produced by the clock-controlled genes (CCGs). The rhythmic oscillation of CCGs is suggested to play an important role in the regulation of circadian activity. However, as we may ascertain, there is a great lack of knowledge regarding the effects of total sleep deprivation (TSD) on neuronal morphology of SCN, and the potential alterations of molecular or neurochemical elements within the circadian system. Based on this viewpoint, the present study was designed to examine (1) the possible ultrastructural variation of SCN and the concurrent change of sodium current after TSD by the use of electron microscopy and time-of-flight secondary ion mass spectrometry (TOF-SIMS) (2) the potential modification of numerous CCGs following TSD under the mRNA and protein levels by the use of RT-PCR and quantitative histochemistry and immunohistochemistry, and (3) the probable beneficial effects of melatonin on conserving the circadian function given that melatonin is a well-established neuroprotective agent. By extensively conduct the present study, we can not only provide the detailed morphological, molecular and biochemical evidences underlying the detrimental effects of TSD on circadian function, but also greatly increase our knowledge on clinical use of melatonin for preserving the normal circadian activity. With the gradually processing of this three-year project, the novel finding acquired will help us to better understand the molecular mechanisms of circadian regulation. In addition, our results will also contribute to the development of sleep medicine and offer the possibility for therapeutic use of melatonin in improving or treating the TSD related circadian disability.慢性睡眠不足或睡眠剝奪為現今工商社會中，廣泛存在影響人類健康的重要問題。 睡眠缺乏將導致腦部功能及生理行為發生改變，包括許多重要酵素暨神經傳導物質的減 少、神經興奮性(neuronal excitability) 受損或約日週期(circadian rhythmicity) 失去平 衡。先前的研究已證實，哺乳類約日週期調控中樞(生物時鐘) 位於前下丘腦(anterior hypothalamus) 之視交叉上核(suprachiasmatic nucleus, SCN)。視交叉上核神經元具有以 24 小時為振盪週期之自發性活性(spontaneous electrical activity)，且此活性由鈉離子流 (sodium ion current) 進出細胞而調節。透過視交叉上核廣泛的投射，此自發性振盪頻率 可直接影響諸多自主神經核的功能運作，包括荷爾蒙的分泌(hormonal secretion) 與睡 眠－清醒週期的運行(sleep-wake cycle operation)。學者研究指出，視交叉上核的神經輸 出訊號由核區內鐘控基因(clock-controlled genes, CCGs) 所合成。鐘控基因的週期性振 盪活性(rhythmic oscillation) 在約日週期的調控機制上，扮演相當重要的角色。然而， 就我們所知，目前學界對睡眠剝奪後，視交叉上核之形態變化幾無所知；另一方面，睡 眠剝奪對約日週期調控系統內，分子與相關神經化學物質活性表現之潛在影響，仍舊付 之闕如。緣此，本計畫預計以三年的時間，逐年依次探討(1) 睡眠剝奪對視交叉上核超 微結構改變暨鈉離子流濃度分布表現之影響。研究擬分別使用電子顯微鏡(electron microscopy) 與飛行式二次離子質譜儀(TOF-SIMS) 分別觀察相關形態暨分子影像的表 現(2) 結合RT-PCR 與定量組織化學、定量免疫組織化學法，分別在mRNA 及蛋白層 次上，深入偵測約日週期調控系統內數種重要鐘控基因蛋白的活性表現，藉以闡明睡眠 剝奪對神經化學運作機制的影響，與(3) 利用褪黑激素(melatonin) 注射，詳加釐清褪 黑激素是否具保存(conserve) 約日週期恆定之神經保護功能(neuroprotective effects)。 藉由此計畫的逐年實行，不僅可以提供學界清晰且詳實有關睡眠剝奪破壞約日週期調控 的形態學、分子影像學暨細胞生化學證據，同時尚對褪黑激素於保護約日週期運作的功 能層面上有所了解。我們希望透過此三年期計畫的順利推動與執行，能有效增進學界對 約日週期調控機制之認識，同時亦對臨床上使用褪黑激素作為預防或治療因睡眠剝奪導 致的週期失調病變(sleep related circadian disorder) 有所幫助。睡眠剝奪視交叉上核約日節律褪黑激素分子影像超微結構定量生化分析Sleep deprivationSuprachiasmatic nucleusCircadian rhythmMelatoninMolecular imageUltrastructureBiochemistryProject