In vivo dual color imaging of neuronal networks during anesthesia

来自家长奖 我们在药理学中面临的剩余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) 和活动(固定)以及痛感丧失(止痛)和对事件的回忆 (失忆)。据认为，GAS通过神经细胞膜上的多种特异性蛋白发挥作用，并 不同的配基门控和电压门控离子通道受到了广泛的关注。其中一个 研究麻醉机制中电压门控钙通道(VGCC)的充分理由 作用是这些通道在调节突触传递和兴奋性方面是必不可少的。 神经元睡眠通路(如丘脑)和参与学习/记忆的大脑区域(如河马)。 坎普·阿尔地层)。重要的是，我们之前的研究已经确定了低电压激活的亚型 VGCC或T型钙通道(T通道)在临床上被不同种类的气体抑制。 相应的浓度范围。在过去的二十年里，我们的工作确立了T-家庭的作用- VGCC用于急慢性疼痛处理，包括手术后疼痛。然而，VGCC的作用 在GA诱导催眠和健忘的机制中，人们仍然难以捉摸。此外，尽管有大量的 在过去的二十年里，我们在理解气体在地球上的行为方式方面取得了进展 分子水平上，关于气体如何在完整水平上引起催眠和记忆障碍的了解要少得多。 神经网络。因此，这项建议旨在阐明特定亚型T通道的贡献 对丘脑皮质(研究区域1)和河马-CampAl环路(研究区域2)的麻醉效应。 我们将利用小鼠的遗传学，在体外选择性地击倒不同的T通道亚型和 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现存教条 并将重点转移到被低估的目标上，例如神经元T通道。我们假设这种理解 GAS靶向T通道的神经生理机制可作为切入点 开发临床麻醉中可能更安全的新方法和做法。米拉机制是 非常适合于实现我们的既定目标，因为我们可以灵活地在研究领域内寻求新的途径 NIGMS感兴趣的。我们实验室始终如一的生产力，以及我们在以下领域与其他人合作的能力 麻醉药理学强烈表明我们的方法将是卓有成效的。建议的工作具有创新性。 在全麻药有用临床作用的新机制中，如意识丧失和 将以健忘症为特征。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在临床麻醉中潜在的更安全的实践。

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.