Investigating non-canonical mechanisms of endogenous opioids on motivation in dorsal midbrain

研究内源性阿片类药物对背侧中脑动机的非典型机制

• 批准号: 10624699
• 负责人: Daniel Charles Castro
• 金额: $ 54.16万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-09 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10624699
• 关键词: Adult; Affect; Affective; Analgesics; Anatomy; Behavior; Behavioral; Biological; Bite; Brain; CRISPR/Cas technology; Calcium; Cell Nucleus; Characteristics; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Color; Data; Desire for food; Devices; Dissociation; Dorsal; Eating; Electric Stimulation; Endoscopy; Enkephalin Receptors; Enkephalins; Fluorescent in Situ Hybridization; Genetic; Goals; Health Care Costs; Image; Individual; Lateral; Ligands; Measures; Mediating; Mental disorders; Microinjections; Midbrain structure; Moods; Morphine; Motivation; Mus; National Institute of Mental Health; Neurons; Nucleus Accumbens; Opioid; Opioid Peptide; Opioid Receptor; Opioid agonist; Opsin; Pattern; Peptides; Pharmacology; Phenotype; Photons; Pilot Projects; Play; Process; Regulation; Rewards; Role; Signal Transduction; Site; Strategic Planning; System; Tail; Technology; Testing; Therapeutic Intervention; United States; Virus; antagonist; behavior test; biological adaptation to stress; calcium indicator; cell type; delta opioid receptor; design; dorsal raphe nucleus; endogenous opioids; experience; experimental study; fluorophore; follow-up; in vivo imaging; increased appetite; insight; knock-down; midbrain central gray substance; motivated behavior; mu receptors; neural; neural circuit; neurobiological mechanism; neurochemistry; neuropsychiatric disorder; neuropsychiatry; neuroregulation; next generation; optogenetics; pharmacologic; receptor; receptor expression; redshift; response; targeted treatment; wireless; wireless implant

Project Summary The primary goal of this R01 is to determine the computational and functional role of endogenous opioids in specific dorsal midbrain nuclei on motivated behaviors. The preponderance of mental illness in the United States results in tens of millions of dollars in healthcare costs. While many neuropsychiatric conditions can be dissociated based on the presence or absence of specific features, a common theme across mental illnesses is the dysregulation of affective or motivated behaviors. The endogenous opioid system is known to powerfully modulate affective and motivational neural circuits. Historically, dorsal midbrain nuclei (including ventrolateral periaqueductal gray nucleus and the adjacent dorsal raphe nucleus) have been shown to be important sites for opioid action. More recently, the lateral dorsal raphe nucleus subregion (LDRN) and nucleus accumbens were shown to be important sites in an opioid-mediated mesolimbic circuit of appetitive motivation. However, while downstream opioid activity in this LDRNaccumbens circuit specifically enhances appetitive motivation, convergent studies indicate that endogenous opioids may play a motivationally suppressive role within LDRN itself. For example, experiments in the 1980s demonstrated morphine microinjections into ventrolateral PAG/LDRN could suppress food intake. Correspondingly, pilot studies in our lab using a CRISPR-Cas9 mediated knockdown of opioid peptides oppositely facilitated food intake. Furthermore, local LDRN opioid activity appears to suppress both appetitive and aversive motivated behaviors (e.g., local opioid antagonism in LDRN increases defensive or escape behaviors). These broad, anti-motivational opioid effects suggest that dysregulation could affect a wide range of affective or motivated behaviors. Therefore, the goal of this R01 application is to determine how endogenous opioids regulate appetitive and aversive motivated behaviors in LDRN by multiplexing genetic, pharmacological, in vivo imaging, and optogenetic technologies. First, we will identify the anatomical characteristics of opioids within dorsal midbrain nuclei (Aim 1). In tandem we will test the functional localization of opioids by performing receptor selective pharmacological antagonism via wireless fluidic devices and CRISPR-Cas9 knockdown of opioid peptides. Next, we will use dual-color 1-photon endoscopic imaging to examine how local opioidergic and non-opioidergic neurons interact to encode appetitive and aversive behaviors (Aim 2). Follow up experiments will use simultaneous 1-photon imaging with cell-type selective optogenetic neuromodulation to augment or disrupt LDRN encoding and expression of motivated behaviors. Finally, we will multiplex 1-photon imaging, CRISPR-Cas9 knockdown, and cell-type specific optogenetic stimulation to determine how endogenous opioid peptides casually augment LDRN encoding and expression of motivated behaviors (Aim 3). Together, these studies may provide insights into how neuropsychiatric disorders are often characterized by affective and motivational dysregulation, and suggest specific neurochemical targets for therapeutic intervention.

项目摘要 R 01的主要目标是确定内源性阿片类药物在脑内的计算和功能作用。 特定的中脑背核对动机行为的影响。美国精神疾病的优势 导致数千万美元的医疗费用。虽然许多神经精神疾病可以 基于特定特征的存在或不存在而分离，精神疾病的共同主题是 情感或动机行为的失调。已知内源性阿片样物质系统 调节情感和动机神经回路历史上，背侧中脑核（包括腹外侧核） 中脑导水管周围灰质核和邻近的中缝背核）已被证明是重要的网站， 阿片类药物作用。最近，中缝背外侧核亚区（LDRN）和背侧核被发现， 被证明是阿片类药物介导的食欲动机的中脑边缘回路中的重要位点。虽然 该LDRN神经回路中的下游阿片样物质活性特异性地增强食欲动机， 聚合研究表明，内源性阿片类物质可能在LDRN中发挥动机抑制作用 本身例如，20世纪80年代的实验表明， PAG/LDRN可抑制摄食。相应地，我们实验室中使用CRISPR-Cas9介导的 阿片肽的敲除相反地促进食物摄取。此外，局部LDRN阿片样物质活性出现 为了抑制食欲和厌恶的动机行为（例如，LDRN中的局部阿片拮抗作用增加 防御或逃避行为）。这些广泛的，反动机阿片类药物的影响表明，失调可能 影响广泛的情感或动机行为。因此，此R 01应用程序的目标是确定 内源性阿片类物质如何通过多重遗传调节LDRN的食欲和厌恶动机行为， 药理学、体内成像和光遗传学技术。首先，我们将确定 背侧中脑核内阿片类物质的特征（Aim 1）。同时，我们将测试功能本地化 通过无线流体装置进行受体选择性药理学拮抗作用， 阿片肽的CRISPR-Cas9敲低。接下来，我们将使用双色单光子内窥镜成像， 研究局部阿片样物质和非阿片样物质神经元如何相互作用以编码食欲和厌恶行为 (Aim 2）。后续实验将使用具有细胞类型选择性光遗传学的同时1光子成像。 神经调节以增强或破坏LDRN编码和动机行为的表达。最后我们将 多重单光子成像、CRISPR-Cas9敲低和细胞类型特异性光遗传学刺激， 确定内源性阿片肽如何偶然地增加LDRN编码和表达的动机， 行为（目标3）。总之，这些研究可能会提供关于神经精神疾病如何经常 其特征是情感和动机失调，并建议特定的神经化学靶点， 治疗干预