Brain Wide Anesthetic-Active Neuronal Network

全脑麻醉活性神经元网络

• 批准号: 10712033
• 负责人: Max Kelz
• 金额: $ 55.78万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712033
• 关键词: 3-Dimensional; Affinity; Anatomy; Anesthesia procedures; Anesthesiology; Anesthetics; Area; Arousal; Behavior assessment; Behavioral; Bioinformatics; Biological; Biological Process; Brain; Brain imaging; Brain region; Caring; Cell Nucleus; Cells; Conscious; Cre driver; Development; Disease; Drug Design; Drug Interactions; Elements; FOS gene; Fiber; Future; General Anesthesia; General anesthetic drugs; Genetic; Goals; Health; Hypnosis; Hypothalamic structure; Immunohistochemistry; Individual; Intervention; Knowledge; Label; Life; Maps; Molecular; Molecular Target; Monitor; Mus; Nervous System; Neurons; Operative Surgical Procedures; Optics; Partner in relationship; Patients; Pattern; Photometry; Physiological; Play; Population; Prefrontal Cortex; Procedures; Property; Reporter; Research; Resolution; Role; Site; Source; Techniques; Thalamic structure; Time; Tissues; Transfection; Transgenic Mice; Transgenic Organisms; Unconscious State; Viral; Virus; Work; analysis pipeline; cell type; clinically significant; connectome; density; designer receptors exclusively activated by designer drugs; druggable target; experience; hypnotic; immunoreactivity; improved; in vivo; innovation; interest; neural circuit; neuronal circuitry; neurophysiology; novel; optogenetics; pharmacologic; rational design; two photon microscopy

Abstract Anesthetics are low affinity drugs that interact with hundreds of molecular targets present throughout the nervous system at clinically significant concentrations. Despite this molecular-level promiscuity, the hypnotic effects of anesthetics depend critically on specific neural circuits. This assertion is supported by numerous results showing that direct modulation of specific sites distributed broadly throughout the brain can potentiate or, conversely, antagonize the anesthetic state. However, because previous experimental work focused upon one brain site at a time, the identification of the long-hypothesized brain-wide canonical anesthesia circuit has so far remained elusive. To fill this critical gap in knowledge, we reasoned that ultimately, the state of anesthesia must be imposed onto the brain by neurons that remain active under anesthesia, while most other neurons are suppressed. To identify anesthetic active neurons throughout the brain, we used tissue clearing and 3D c-Fos immunohistochemistry. We validated that putative anesthesia-active neurons are indeed physiologically active in vivo using two photon microscopy and fiber photometry. Having identified anesthesia- active neurons, we further reasoned that brain regions which project broadly are more likely to play a pivotal role in modulating the level of consciousness. Thus, we combined our brain activity map with whole brain connectivity analyses. The potent combination of these experimental and bioinformatics approaches allowed us to identify regions that contain a high density of anesthetic-active neurons and project broadly throughout the brain. Our unbiased approach culminated in the definition of a putative canonical anesthesia network comprised of nuclei in the ventral hypothalamus, thalamus, and the prefrontal cortex. In Aim 1, we will discover the specific cell types within prefrontal cortex, ventral hypothalamus and thalamus that remain active under anesthesia. This is of critical importance as all brain regions contain many cell subtypes with distinct neurophysiological properties, connectivity patterns, and ultimately, behavioral effects. In Aim 2, we will discover the brain-wide projections made by anesthetic-active neurons by combining anterograde and retrograde viral tracing in transgenic mice that specifically label distinct neuronal subtypes with 3D immunohistochemistry. In Aim 3, we will establish the functional roles of the canonical anesthesia network as a whole and each of its elements individually by combining chemo- and optogenetics with behavioral and neurophysiologic assessments of arousal. The ultimate result of the proposed research will be the identification of a canonical network of neurons sufficient to elicit hypnosis. This has fundamental implications for how the state of arousal is controlled in health and dysregulated in disease. Identification of this circuit may also suggest druggable targets for the development of more specific anesthetic agents with fewer undesirable effects. This has the potential to significantly improve the care of millions of patients requiring general anesthesia for life saving procedures.

摘要 麻醉剂是低亲和力药物，其与存在于全身的数百个分子靶点相互作用。 在临床上显著浓度的神经系统。尽管有这种分子水平的混杂， 麻醉剂的效果主要取决于特定的神经回路。这一说法得到了众多 结果表明，直接调制广泛分布在整个大脑中的特定部位可以增强 或者相反对抗麻醉状态然而，由于以前的实验工作集中在 一次一个大脑部位，对长期假设的全脑典型麻醉回路的识别， 至今仍然难以捉摸。为了填补这一关键的知识空白，我们推断，最终， 麻醉必须通过在麻醉下保持活性的神经元施加到大脑上，而大多数其他神经元必须在麻醉下保持活性。 神经元受到抑制。为了识别整个大脑的麻醉活性神经元，我们使用组织清除 和三维c-Fos免疫组化。我们证实，假定的麻醉活性神经元确实是 使用双光子显微镜和纤维光度法测定在体内的生理活性。已经确定了麻醉- 活跃的神经元，我们进一步推断，大脑中广泛投射的区域更有可能发挥关键作用。 调节意识水平的作用。因此，我们将我们的大脑活动图与整个大脑相结合， 连通性分析。这些实验和生物信息学方法的有效结合， 我们识别出含有高密度麻醉活性神经元的区域， 大脑我们的公正的方法最终在一个公认的规范麻醉网络的定义 由腹侧下丘脑、丘脑和前额皮质的核团组成。在目标1中，我们将发现 前额叶皮层、腹侧下丘脑和丘脑内的特定细胞类型在 麻醉这是至关重要的，因为所有的大脑区域都包含许多具有不同功能的细胞亚型。 神经生理学特性、连接模式以及最终的行为效应。在目标2中，我们将 通过结合顺行和顺行， 用3D特异性标记不同神经元亚型的转基因小鼠中的逆行病毒追踪 免疫组化在目标3中，我们将建立规范麻醉网络的功能角色， 通过将化学和光遗传学与行为和 神经生理学评估拟议研究的最终结果将是确定 一个典型的神经元网络足以引发催眠。这对如何从根本上影响 在健康状态下，唤醒状态受到控制，而在疾病状态下，唤醒状态失调。该电路的标识还可以 提出了开发更特异性的麻醉剂的可药用靶点， 方面的影响.这有可能显着改善数百万需要一般护理的患者的护理。 用于救生手术的麻醉。