Circadian rhythm of PERIOD2::LUCIFERASE expression in the trigeminal ganglion of mice.
Circadian rhythm of PERIOD2::LUCIFERASE expression in the trigeminal ganglion of mice.
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DOI：
10.3389/fnins.2023.1142785
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发表时间：
2023
影响因子：
4.3
通讯作者：
Sugimura, Mitsutaka
中科院分区：
文献类型：
--
作者：
Shirakawa, Yukie;Ohno, Sachi N. N.;Yamagata, Kanae A. A.;Kuramoto, Eriko;Oda, Yoshiaki;Nakamura, Takahiro J. J.;Nakamura, Wataru;Sugimura, Mitsutaka
三叉神经在口腔和面部区域传递温暖、疼痛和触觉压力等敏感感觉，大多数三叉神经传入细胞体位于三叉神经节。我们之前的研究表明，三叉神经支配区域的感觉，特别是在颌面区域，表现出昼夜变化，敏感性随时间变化。在本研究中，我们旨在阐明小鼠三叉神经节生物钟基因的表达节律，以阐明该区域的昼夜节律调节机制。免疫组化检测PER2蛋白在野生型小鼠视交叉上核和三叉神经节中的表达。采用PERIOD2::LUCIFERASE敲入（PER2::LUC）小鼠作为生物发光检测基因表达。体外培养单侧三叉神经节和包括视交叉上核在内的脑切片。然后用高灵敏度的光电探测器测量生物发光水平。然后用Cry1基因缺陷（Cry1−/−）或Cry2基因缺陷（Cry2−/−）小鼠进行相同的实验。在三叉神经节中，免疫组织化学定位了神经元胞体内PER2蛋白的表达。小鼠三叉神经节离体组织中PER2::LUC水平在所有基因型、野生型、Cry1−/−和Cry2−/−中显示出明显的昼夜振荡。三叉神经节的周期短于视交叉上核；Cry1−/−小鼠比野生型小鼠短，Cry2−/−小鼠比野生型小鼠长。Per2在离体培养三叉神经节神经元中的表达和不同昼夜节律的振荡表明，三叉神经节通过自主昼夜节律振荡负责感觉输入和时间门控的中继。
The trigeminal nerve conveys delicate sensations such as warmth, pain, and tactile pressure in the oral and facial regions, and most trigeminal afferent cell bodies are located in the trigeminal ganglion. Our previous study has shown that sensations in trigeminal nerve innervated areas, specifically in the maxillofacial region, exhibit diurnal variation and that sensitivity changes time-dependently. In this study, we aimed to clarify the rhythm of expression of clock gene in the trigeminal ganglion of mice to elucidate the mechanism of circadian regulation in the same area. Immunohistochemistry examined the expression of the PER2 protein in the suprachiasmatic nucleus and trigeminal ganglion of wild-type mice. To measure gene expression as bioluminescence, PERIOD2::LUCIFERASE knock-in (PER2::LUC) mice were used. Unilateral trigeminal ganglion and brain sections including the suprachiasmatic nucleus were incubated ex vivo. Bioluminescence levels were then measured using a highly sensitive photodetector. The same experiments were then conducted with Cry1 gene-deficient (Cry1−/−) or Cry2 gene-deficient (Cry2−/−) mice. In the trigeminal ganglion, immunohistochemistry localized PER2 protein expression within the neuronal cell body. Mouse trigeminal ganglion ex vivo tissues showed distinct circadian oscillations in PER2::LUC levels in all genotypes, wild-type, Cry1−/−, and Cry2−/−. The period was shorter in the trigeminal ganglion than in the suprachiasmatic nucleus; it was shorter in Cry1−/− and longer in Cry2−/− mice than in the wild-type mice. The expression of Per2 in neurons of the trigeminal ganglion in ex vivo culture and the oscillation in a distinct circadian rhythm suggests that the trigeminal ganglion is responsible for the relay of sensory inputs and temporal gating through autonomous circadian oscillations.
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