Neuroscience and Mental Health Institute, University of Alberta, Edmonton, Canada.
Neurochemical Research Unit, University of Alberta, Edmonton, Canada.
Elife. 2024 Apr 30;12:RP90080. doi: 10.7554/eLife.90080.
Despite substantial progress in mapping the trajectory of network plasticity resulting from focal ischemic stroke, the extent and nature of changes in neuronal excitability and activity within the peri-infarct cortex of mice remains poorly defined. Most of the available data have been acquired from anesthetized animals, acute tissue slices, or infer changes in excitability from immunoassays on extracted tissue, and thus may not reflect cortical activity dynamics in the intact cortex of an awake animal. Here, in vivo two-photon calcium imaging in awake, behaving mice was used to longitudinally track cortical activity, network functional connectivity, and neural assembly architecture for 2 months following photothrombotic stroke targeting the forelimb somatosensory cortex. Sensorimotor recovery was tracked over the weeks following stroke, allowing us to relate network changes to behavior. Our data revealed spatially restricted but long-lasting alterations in somatosensory neural network function and connectivity. Specifically, we demonstrate significant and long-lasting disruptions in neural assembly architecture concurrent with a deficit in functional connectivity between individual neurons. Reductions in neuronal spiking in peri-infarct cortex were transient but predictive of impairment in skilled locomotion measured in the tapered beam task. Notably, altered neural networks were highly localized, with assembly architecture and neural connectivity relatively unaltered a short distance from the peri-infarct cortex, even in regions within 'remapped' forelimb functional representations identified using mesoscale imaging with anaesthetized preparations 8 weeks after stroke. Thus, using longitudinal two-photon microscopy in awake animals, these data show a complex spatiotemporal relationship between peri-infarct neuronal network function and behavioral recovery. Moreover, the data highlight an apparent disconnect between dramatic functional remapping identified using strong sensory stimulation in anaesthetized mice compared to more subtle and spatially restricted changes in individual neuron and local network function in awake mice during stroke recovery.
尽管在描绘局灶性缺血性中风引起的网络可塑性轨迹方面取得了重大进展,但在小鼠梗死周边皮质内神经元兴奋性和活动的变化程度和性质仍未得到明确界定。大多数现有数据是从麻醉动物、急性组织切片或从提取组织的免疫测定推断兴奋性变化获得的,因此可能无法反映清醒动物完整皮质中的皮质活动动态。在这里,使用清醒、行为正常的小鼠体内双光子钙成像技术,在光血栓性中风靶向前肢体感皮层后 2 个月内,对皮质活动、网络功能连接和神经组装结构进行纵向跟踪。在中风后的几周内跟踪感觉运动恢复情况,使我们能够将网络变化与行为联系起来。我们的数据显示体感神经网络功能和连接存在空间限制但持久的改变。具体来说,我们证明了在个体神经元之间的功能连接出现缺陷的同时,神经组装结构也出现了显著且持久的破坏。在梗死周边皮质中的神经元放电减少是短暂的,但与在锥形束任务中测量的熟练运动障碍相关。值得注意的是,改变的神经网络高度局限,即使在中风后 8 周使用麻醉制剂的介观成像识别的“重新映射”前肢功能代表区域内,组装结构和神经连接也相对不变。因此,使用清醒动物的纵向双光子显微镜,这些数据显示了梗死周边神经元网络功能与行为恢复之间的复杂时空关系。此外,数据突出表明,在麻醉小鼠中使用强烈的感觉刺激识别出的戏剧性功能重映射与清醒小鼠中风恢复期间单个神经元和局部网络功能的更微妙和空间限制变化之间存在明显的脱节。