In vivo dual color imaging of neuronal networks during anesthesia

家长奖 我们在药理学中面临的剩下的基本挑战之一是破译 全身麻醉药（GAs）的作用。完全麻醉状态包括意识丧失（催眠） 和运动（固定），以及痛觉丧失（镇痛）和对事件的回忆 （健忘症）。据信，GAs通过神经元膜上的多种但特异的蛋白质起作用， 不同的配体门控和电压门控离子通道已经受到了重要的考虑。之一 研究电压门控钙通道（VGCC）在麻醉机制中的作用 这些通道在调节突触传递和兴奋性中是必不可少的。 神经元睡眠通路（例如丘脑）和参与学习/记忆的脑区域（例如河马- 形成）。重要的是，我们以前的研究已经确定，低电压激活的亚型， VGCC或T型钙通道（T通道）在临床上被不同种类的GA抑制。 相关浓度范围。在过去的二十年里，我们的工作确立了T家族的作用- 型VGCC在急性和慢性疼痛处理中，包括手术后疼痛。然而，自愿捐助国的作用 在GA诱导的催眠和健忘症的机制仍然是难以捉摸的。此外，尽管大量 在过去的二十年里，我们在理解遗传算法如何在 分子水平上，更少的是知道如何GA导致催眠和记忆缺陷的水平上完整的 神经网络因此，该建议旨在阐明T通道的特定亚型的贡献 对丘脑皮质（研究区域1）和海马神经回路（研究区域2）的麻醉作用。 我们将利用小鼠遗传学，体外选择性敲低不同的T通道亚型， 体内电生理学、光学记录以及一系列行为测试来解决这些关键问题。 挑战我们提出的工作有可能推翻关于麻醉机制的过去教条 并将重点转移到未被充分认识的目标，如神经元T通道。我们尊重这种理解 靶向T通道的GA的神经生理学作用机制可以用作起点 在临床麻醉中开发新的和潜在的更安全的方法和实践。MIRA机制是 非常适合实现我们的既定目标，因为在研究领域内寻求新途径的灵活性 对NIGMS感兴趣。我们实验室的稳定生产力以及我们与该领域其他人合作的能力 麻醉药理学强有力地表明我们的方法将是富有成效的。建议的工作是创新的 在全身麻醉药有用临床效果的新机制中， 健忘症的特征。它具有医学意义，因为它描述了药物的重要性， 靶向电压门控钙通道，用于临床麻醉中更安全实践的潜在发展。

From Parent Award One of the remaining fundamental challenges we face in pharmacology is deciphering the mechanisms of action of general anesthetics (GAs). A complete anesthetic state involves loss of consciousness (hypnosis) and movement (immobilization), as well as loss of pain sensation (analgesia) and recollection of the event (amnesia). It is believed that GAs act through the multiple but specific proteins on neuronal membrane and different ligand-gated and voltage-gated ion channels have received a significant consideration. One of the compelling reasons to study voltage-gated calcium channels (VGCCs) in the mechanisms of anesthetic actions is that these channels are essential in regulation of synaptic transmission and excitability in the neuronal sleep pathway (e.g. thalamus) and in the brain regions involved in learning/memory (e.g. hippo- campal formation). Importantly, our previous studies have established that low-voltage-activated subtype of VGCCs or T-type calcium channels (T-channels) are inhibited by different classes of GAs within the clinically relevant concentration range. For the past two decades our work has established the role of the family of T- type VGCCs in acute and chronic pain processing, including post-surgical pain. However the role of VGCCs in the mechanisms of GA-induced hypnosis and amnesia remains elusive. Furthermore, despite substantial progress that has been made in the last two decades towards our understanding of how GAs act at the molecular level, much less is known about how GAs cause hypnosis and memory deficit at the level of intact neuronal networks. Hence, this proposal aims to elucidate the contribution of specific subtypes of T-channels to anesthetic effects in the thalamocortical (Research area 1) and hippo- campal circuitry (Research area 2). We will take advantage of mouse genetics, selective knock-down of different T-channel isoforms ex vivo and in vivo electrophysiology, optical recordings, as well as a battery of behavioral tests to address these key challenges. Our proposed work has the potential to overturn ex- isting dogma about anesthetic mechanisms and to shift the focus to underappreciated targets, such as neuronal T-channels. We posit that understanding the neurophysiological mechanisms of action of GAs that target T-channels may be used as a starting point to develop novel and potentially safer approaches and practices in clinical anesthesia. MIRA mechanism is well suited to achieve our stated goals because of flexibility to pursue new avenues within the research area of interest to NIGMS. Consistent productivity of our lab and our ability to collaborate with others in the field of anesthetic pharmacology strongly suggest that our approach will be fruitful. The proposed work is innovative in that new mechanisms of useful clinical effects of general anesthetics such as loss of consciousness and amnesia will be characterized. It is med- ically significant because it describes the importance of drugs that target voltage-gated calcium channels for potential development of safer practices in clinical anesthesia.