日本东京大学Hiroki R. Ueda团队发现，突触后钙调神经磷酸酶和蛋白激酶A的竞争调节哺乳动物的睡眠-觉醒周期。该研究于2024年11月6日在线发表于国际一流学术期刊《自然》。

研究人员探讨了一对互相调节睡眠持续时间的激酶和磷酸酶。首先，通过使用产前和产后CRISPR靶向技术，研究人员进行蛋白激酶A（PKA）和磷蛋白磷酸酶（PPP）家族基因的全面筛选，并生成40个基因敲除小鼠模型。

研究人员鉴定出了PKA的调节亚基（Prkar2b）、蛋白磷酸酶1（PP1）的调节亚基（Pppr1r9b）以及钙调神经磷酸酶（也称为PP2B）的催化和调节亚基（Ppp3ca和Ppp3r1）作为睡眠控制基因。

通过腺相关病毒（AAV）介导的PKA和PP1-钙调神经磷酸酶活性刺激，研究人员发现PKA是促进觉醒的激酶，而PP1和钙调神经磷酸酶则作为促进睡眠的磷酸酶。

睡眠调控中这些磷酸酶的重要性，得到了其活性增加或减少所引起的睡眠持续时间显著变化的支持，变化范围从大约17.3小时/天（PP1表达）到4.3小时/天（钙调神经磷酸酶的产后CRISPR靶向）。、

这些磷酸酶的定位信号需要作用于兴奋性突触后膜，才能发挥其促进睡眠的效应。此外，定位于兴奋性突触后膜的PKA的促进觉醒效应，抵消了PP1-钙调神经磷酸酶的促进睡眠效应。这些发现表明，PKA和PP1-钙调神经磷酸酶在兴奋性突触后膜上具有竞争性作用，共同调节睡眠。

据悉，突突触蛋白的磷酸化，是调控哺乳动物睡眠-觉醒周期的重要生化反应。体内的蛋白质磷酸化，是由激酶和磷酸酶可逆调节的。

附：英文原文

Title: Postsynaptic competition between calcineurin and PKA regulates mammalian sleep–wake cycles

Author: Wang, Yimeng, Cao, Siyu, Tone, Daisuke, Fujishima, Hiroshi, Yamada, Rikuhiro G., Ohno, Rei-ichiro, Shi, Shoi, Matsuzawa, Kyoko, Yada, Saori, Kaneko, Mari, Sakamoto, Hirokazu, Onishi, Taichi, Ukai-Tadenuma, Maki, Ukai, Hideki, Hanashima, Carina, Hirose, Kenzo, Kiyonari, Hiroshi, Sumiyama, Kenta, Ode, Koji L., Ueda, Hiroki R.

Issue&Volume: 2024-11-06

Abstract: The phosphorylation of synaptic proteins is a significant biochemical reaction that controls the sleep–wake cycle in mammals1,2,3. Protein phosphorylation in vivo is reversibly regulated by kinases and phosphatases. In this study, we investigate a pair of kinases and phosphatases that reciprocally regulate sleep duration. First, we perform a comprehensive screen of protein kinaseA (PKA) and phosphoprotein phosphatase (PPP) family genes by generating 40 gene knockout mouse lines using prenatal and postnatal CRISPR targeting. We identify a regulatory subunit of PKA (Prkar2b), a regulatory subunit of protein phosphatase1 (PP1; Pppr1r9b) and catalytic and regulatory subunits of calcineurin (also known as PP2B) (Ppp3ca and Ppp3r1) as sleep control genes. Using adeno-associated virus (AAV)-mediated stimulation of PKA and PP1–calcineurin activities, we show that PKA is a wake-promoting kinase, whereas PP1 and calcineurin function as sleep-promoting phosphatases. The importance of these phosphatases in sleep regulation is supported by the marked changes in sleep duration associated with their increased and decreased activities, ranging from approximately 17.3h per day (PP1 expression) to 4.3h per day (postnatal CRISPR targeting of calcineurin). Localization signals to the excitatory post-synapse are necessary for these phosphatases to exert their sleep-promoting effects. Furthermore, the wake-promoting effect of PKA localized to the excitatory post-synapse negated the sleep-promoting effect of PP1–calcineurin. These findings indicate that PKA and PP1–calcineurin have competing functions in sleep regulation at excitatory post-synapses.

DOI: 10.1038/s41586-024-08132-2

Source: https://www.nature.com/articles/s41586-024-08132-2