In Vivo Photoadduction of Anesthetic Ligands in Mouse Brain Markedly Extends Sedation and Hypnosis.
In Vivo Photoadduction of Anesthetic Ligands in Mouse Brain Markedly Extends Sedation and Hypnosis.
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DOI：
10.1523/jneurosci.1884-22.2023
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发表时间：
2023-03-29
影响因子：
5.3
通讯作者：
Kelz, Max B.
中科院分区：
文献类型：
--
作者：
McKinstry-Wu, Andrew R.;Wasilczuk, Andrzej Z.;Dailey, William P.;Eckenhoff, Roderic G.;Kelz, Max B.
光亲和配体最为人所知的是用于鉴定药物与其分子靶标的特异性结合位点的工具。然而，光亲和配体具有进一步确定药物作用的关键神经解剖学靶点的潜力。在野生型雄性小鼠的大脑中，我们证明了在体内使用光亲和配体的可行性，通过靶向但空间限制的azi-m-propofol（aziPm），全身麻醉剂丙泊酚的光反应类似物的光加合来延长麻醉。与没有紫外线照射的对照小鼠相比，在臂旁核和蓝斑的边界处，在脑桥的嘴侧进行双侧近紫外线光内收的aziPm的全身给药产生了20倍的镇静和催眠作用的持续时间增加。错过臂旁蓝斑复合体的光内收也未能延长aziPm的镇静或催眠作用，并且与非内收对照组无法区分。为了证实体内靶向光内收的长期行为和EEG结果，我们在脑桥脑片的嘴侧进行了电生理记录。使用蓝斑内的神经元，以进一步突出不可逆aziPm结合的细胞后果，我们证明了短暂的洗澡应用aziPm，成为不可逆的光内收的自发动作电位的瞬时减慢。总之，这些研究结果表明，光化学为基础的战略是一个可行的新方法探测中枢神经系统的生理和病理生理。光亲和配体是能够光诱导不可逆结合的药物，其在确定药物作用的神经解剖学位点方面具有未开发的潜力。我们在小鼠中全身给予中枢作用的麻醉剂光亲和配体，在脑内进行局部光照射以在其体内作用部位共价加合药物，并成功地在有限的250 µm半径内富集不可逆的药物结合。当光内收包括脑桥臂旁蓝斑复合体，麻醉镇静和催眠延长20倍，从而说明了在体内光化学的力量，以帮助解开药物作用的神经元机制。
Photoaffinity ligands are best known as tools used to identify the specific binding sites of drugs to their molecular targets. However, photoaffinity ligands have the potential to further define critical neuroanatomic targets of drug action. In the brains of WT male mice, we demonstrate the feasibility of using photoaffinity ligands in vivo to prolong anesthesia via targeted yet spatially restricted photoadduction of azi-m-propofol (aziPm), a photoreactive analog of the general anesthetic propofol. Systemic administration of aziPm with bilateral near-ultraviolet photoadduction in the rostral pons, at the border of the parabrachial nucleus and locus coeruleus, produced a 20-fold increase in the duration of sedative and hypnotic effects compared with control mice without UV illumination. Photoadduction that missed the parabrachial-coerulean complex also failed to extend the sedative or hypnotic actions of aziPm and was indistinguishable from nonadducted controls. Paralleling the prolonged behavioral and EEG consequences of on target in vivo photoadduction, we conducted electrophysiologic recordings in rostral pontine brain slices. Using neurons within the locus coeruleus to further highlight the cellular consequences of irreversible aziPm binding, we demonstrate transient slowing of spontaneous action potentials with a brief bath application of aziPm that becomes irreversible on photoadduction. Together, these findings suggest that photochemistry-based strategies are a viable new approach for probing CNS physiology and pathophysiology. SIGNIFICANCE STATEMENT Photoaffinity ligands are drugs capable of light-induced irreversible binding, which have unexploited potential to identify the neuroanatomic sites of drug action. We systemically administer a centrally acting anesthetic photoaffinity ligand in mice, conduct localized photoillumination within the brain to covalently adduct the drug at its in vivo sites of action, and successfully enrich irreversible drug binding within a restricted 250 µm radius. When photoadduction encompassed the pontine parabrachial-coerulean complex, anesthetic sedation and hypnosis was prolonged 20-fold, thus illustrating the power of in vivo photochemistry to help unravel neuronal mechanisms of drug action.
影响因子：
3
作者：
Wasilczuk AZ;Meng QC;McKinstry-Wu AR
通讯作者：
McKinstry-Wu AR
11-三氟甲基 - 苯基二氮嗪类神经类似类似物：GABAA受体的有效的一般麻醉和光标记试剂。
影响因子：
3.4
作者：
Chen, Zi-Wei;Wang, Cunde;Krishnan, Kathiresan;Manion, Brad D.;Hastings, Randy;Bracamontes, John;Taylor, Amanda;Eaton, Megan M.;Zorumski, Charles F.;Steinbach, Joseph H.;Akk, Gustav;Mennerick, Steven;Covey, Douglas F.;Evers, Alex S.
通讯作者：
Evers, Alex S.