目的:因为在不同的大脑区域,GABA神经元的活性在很大程度上是未知的,我们测量了在无核带(ZI)和腹侧导水管周围灰色(vlPAG)的GABA神经元中钙荧光的体内变化,与调节睡眠有关的两个方面。
方法:vGAT-Cre小鼠植入睡眠电极,将rAAV-DIO-GCaMP6显微注射入ZI(n=6)或vlPAG(n=5)(异氟烷麻醉),并将GRIN晶状体插入注射部位。21天后,微镜在多个REM周期中记录了单个vGAT神经元的荧光。用PCA-ICA分析自动提取对应于个体vGAT躯体的感兴趣区域。
结果:在ZI中,鉴定372个神经元。以前,我们记录了ZI中310个vGAT神经元的活性(Blanco-Centurion等人。,2021年),我们将已发布的数据集与新数据集相结合,创建了ZIvGAT神经元的综合数据集(总神经元=682;小鼠=11)。在vlPAG中,鉴定了169个神经元(小鼠=5)。在这两个区域中,大多数神经元在REM睡眠中都处于最大活跃状态(R-Max;ZI=51.0%,vlPAG=60.9%)。第二丰富的组是W-Max(ZI=23.9%,vlPAG=25.4%)。在ZI中,但不是在vlPAG中,神经元为NREMS-Max(11.7%)。vlPAG具有REMS-Off神经元(8.3%)。在这两个领域都有两个小类:wake/REMS-Max和状态冷漠。在ZI中,NREMS-Max神经元在睡眠开始前30秒发出荧光。
结论:这些描述性数据表明,在与睡眠有关的两个大脑区域中,GABA神经元的活性偏向于睡眠。
As in various brain regions the activity of gamma-aminobutyric acid (GABA) neurons is largely unknown, we measured in vivo changes in calcium fluorescence in GABA neurons in the zona incerta (ZI) and the ventral lateral periaqueductal grey (vlPAG), two areas that have been implicated in regulating sleep.
vGAT-Cre mice were implanted with sleep electrodes, microinjected with rAAV-DIO-
GCaMP6 into the ZI (n = 6) or vlPAG (n = 5) (isoflurane anesthesia) and a GRIN (Gradient-Index) lens inserted atop the injection site. Twenty-one days later, fluorescence in individual vGAT neurons was recorded over multiple REM cycles. Regions of interest corresponding to individual vGAT somata were automatically extracted with PCA-ICA analysis.
In the ZI, 372 neurons were identified. Previously, we had recorded the activity of 310 vGAT neurons in the ZI and we combined the published dataset with the new dataset to create a comprehensive dataset of ZI vGAT neurons (total neurons = 682; mice = 11). In the vlPAG, 169 neurons (mice = 5) were identified. In both regions, most neurons were maximally active in REM sleep (R-Max; ZI = 51.0%, vlPAG = 60.9%). The second most abundant group was W-Max (ZI = 23.9%, vlPAG = 25.4%). In the ZI, but not in vlPAG, there were neurons that were NREMS-Max (11.7%). vlPAG had REMS-Off neurons (8.3%). In both areas, there were two minor classes: wake/REMS-Max and state indifferent. In the ZI, the NREMS-Max neurons fluoresced 30 s ahead of sleep onset.
These descriptive data show that the activity of GABA neurons is biased in favor of sleep in two brain regions implicated in sleep.