Neuronal Activity in Sleep & Wake in Alzheimer's Disease Mice

深部脑成像方法(显微内窥镜)是一种高通量的方法，可以识别 神经元特定表型的活动的时间和空间模式。我们假设过 小网络中的活动可以识别正常大脑和疾病大脑。为了检验这一假设，我们将使用 双波长微型望远镜成像5xFAD小鼠部分神经元活性的实验研究 阿尔茨海默病模型(正常对照纯合子小鼠)。我们创造了发起人- 驱动钙传感器，驱动红移的钙指示器jRGECO进入食欲素唤醒 神经元。GABA睡眠神经元中的一种单独的钙传感器(GCaMP6m)将用作对比。 对两个不同波长的神经元进行成像的优势在于，它允许比较 当大脑在清醒和睡眠之间转换时，唤醒和睡眠神经元。我们将为 在正常清醒-睡眠期间和睡眠丧失6小时后，含有jRGECO和GCaMP的神经元。我们 假设在从尾迹过渡过程中存在异常的荧光时间模式 在5xFAD小鼠的大脑中睡觉，随着疾病的发展，这种情况会恶化。这可能解释了 阿尔茨海默病睡眠-觉醒模式紊乱的原因。我们假设异常活动 在大脑深层回路中，调节睡眠是疾病的先兆。这一小笔预算的总体影响 该项目将识别觉醒过程中两个不同并列的神经元NREM和NREM的活动 在野生型和疾病模型中的REM睡眠，这将有助于理解小簇 当清醒的大脑进入睡眠状态时，神经元就会发挥作用。

The deep-brain imaging approach (microendoscopy) is a high throughput method that identifies the temporal and spatial pattern of activity in specific phenotypes of neurons. We have hypothesized that activity in small networks can identify normal versus diseased brains. To test this hypothesis, we will use the two-wavelength miniscope to image activity of a subset of neurons in 5xFAD mice, a validated animal model of Alzheimer’s disease (normal versus the homozygous littermates). We have created promoter- driven calcium sensor that drives the red-shifted calcium indicator, jRGECO, into orexin-arousal neurons. A separate calcium sensor (GCaMP6m) in GABA sleep neurons will be used as comparison. The advantage of imaging neurons with two different wavelengths is that it allows for comparison of arousal versus sleep neurons as the brain transitions between wake and sleep. We will image activity of neurons that contain jRGECO and GCaMP during normal wake-sleep bouts and after 6h sleep loss. We hypothesize that there is an abnormal temporal pattern of fluorescence during the transition from wake to sleep in the brains of 5xFAD mice, and it worsens with progression of disease. This may explain the cause of the disrupted sleep-wake patterns in Alzheimer’s disease. We hypothesize that abnormal activity in deep brain circuits regulating sleep is a harbinger of disease. The overall impact of this small budget project is that it will identify activity of two different juxtapositioned neurons during wake, NREM and REM sleep in wildtype versus a disease model, which will aid in understanding how small clusters of neurons behave as the waking brain falls asleep.