IMAGING EPILEPTIFORM EVENTS IN JUVENILE NEOCORTEX

描述：(摘自申请者的摘要)约2%的人患有癫痫 这一现象在世界人口中占有重要地位，尤其是在儿童中。相对的 癫痫发作是如何传播和招募的知之甚少 显然是正常的大脑皮层回路。考虑到大脑皮层的复杂性， 其中涉及数十类兴奋性和抑制性神经元 不同的电路功能，很可能是引发和传播 癫痫放电受特定神经元的不同控制 上课。在过去的十年里，我们开发了一种光学方法，使用 从神经元群体中进行钙成像以研究新皮质回路和 想象它们在双光子激发下在三维空间的激活。vbl.使用 这一策略，我们可以光学地检测到来自 数十或数百个神经元，用单个细胞成像癫痫样事件 分辨并检测哪些神经元参与不同类型的 癫痫样事件。我们建议进行系统的努力，以了解 不同类型的新皮质神经元在中枢神经系统中的起始和传播 癫痫。我们将使用钙成像的神经细胞群体在 青少年大脑新皮质脑片药物致痫样事件 (P9-P20)大鼠体感觉皮层，以便更好地了解回路 青少年癫痫的发病机制，同时也为青少年癫痫的 从发作间歇期过渡到发作间歇期。这些实验将会进行 将全细胞记录和生物细胞素重建与 最先进的成像技术，包括双光子显微镜， 光电二极管阵列和快速冷却的ccd摄像机。我们的第一个目标是 从形态和生理上描述参与其中的神经元 自发和诱发的发作间期和发作期癫痫样事件。我们的最终目标 是应用一种新的光学探测方法来重建电路 通过揭示突触后靶点来揭示癫痫样事件 由第5层IB神经元或其他潜在的关键细胞类触发。 这些问题的答案可能会对 靶向特定的神经元或细胞层，这些神经元或细胞层在 癫痫样事件。此外，我们的结果将在 确定导致下限的蜂窝和电路机制 发育期和幼年期新皮质的惊厥阈值及其转变 发作间歇期至发作间歇期。

DESCRIPTION: (from applicant's abstract) Epilepsy affects about 2 percent of the world population and is particularly frequent in children. Relatively little is known about how epileptic seizures propagate across and recruit apparently normal cortical circuits. Given the complexity of the neocortex, where dozens of classes of excitatory and inhibitory neurons are involved in different circuit functions, it is likely that the initiation and spread of epileptic discharges are differentially controlled by specific neuronal classes. Over the last decade, we have developed an optical approach using calcium imaging from population of neurons to study neocortical circuits and to image their activation in three dimensions with 2 photon excitations. Using this strategy, we can optically detect action potentials in the somata from dozens or hundreds of neurons, image epileptiform events with single cell resolution and detect which neurons participate in different types of epileptiform events. We propose a systematic effort to understand the role of different classes of neocortical neurons in the initiation and propagation of epilepsy. We will use calcium imaging of neuronal populations during pharmacological-induced epileptiform events in neocortical slices from juvenile (P9 - P20) rat somatosensory cortex, in order to better understand the circuit mechanisms responsible for juvenile epilepsy and at the same time image the transition from interictal to ictal events. The experiments will be carried out combining whole cell recordings and biocytin reconstructions with state-of-the-art imaging techniques, including two-photon microscopy, a photodiode array and a fast cooled CCD camera. Our first goal is to characterize morphologically and physiologically the neurons involved in spontaneous and evoked interictal and ictal epileptiform events. Our final goal is to apply a novel optical probing method to reconstruct the circuitry underlying epileptiform events by revealing the postsynaptic targets that are triggered by layer 5 IB neurons, or other potentially key cell classes. The answers to these questions could have therapeutic implications for targeting specific neurons or cell layers which play a critical role in epileptiform events. Also, our results will be particularly useful in identifying the cellular and circuit mechanism responsible for the lower seizure threshold of developing and juvenile neocortex and the transition from interictal to ictal events.