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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要该 R01 的主要目标是确定内源性阿片类药物在计算和功能方面的作用。特定背侧中脑核对动机行为的影响美国精神疾病的患病率。导致数千万美元的医疗费用，而许多神经精神疾病可能会导致。根据特定特征的存在或不存在而分离，精神疾病的一个共同主题是众所周知，内源性阿片系统会导致情感或动机行为的失调。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）。导水管周围灰核和邻近的中缝背核）已被证明是重要的位点最近，外侧中缝背核亚区（LDRN）和伏隔核被研究。已被证明是阿片类药物介导的中脑边缘回路中的重要位点。 LDRNaccumbens 回路中的下游阿片类药物活性特别增强食欲动机，趋同研究表明内源性阿片类药物可能在 LDRN 中发挥动机抑制作用例如，20 世纪 80 年代的实验证明了将吗啡显微注射到腹外侧。相应地，我们实验室使用 CRISPR-Cas9 介导的初步研究表明，PAG/LDRN 可以抑制食物摄入。此外，局部 LDRN 敲除阿片类药物活性出现抑制食欲和厌恶动机的行为（例如，LDRN 中的局部阿片类药物拮抗作用增加这些广泛的、反动机的阿片类药物效应表明，调节失调可能因此，该 R01 应用程序的目标是确定。内源性阿片类药物如何通过多重遗传来调节 LDRN 中的食欲和厌恶动机行为，首先，我们将确定解剖学、体内成像和光遗传学技术。背侧中脑核内阿片类药物的特征（目标 1）同时我们将测试功能定位。通过无线流体装置执行受体选择性药理拮抗作用来治疗阿片类药物接下来，我们将使用双色 1 光子内窥镜成像来敲除阿片肽。检查局部阿片能和非阿片能神经元如何相互作用以编码食欲和厌恶行为（目标 2）后续实验将使用同步 1 光子成像和细胞类型选择性光遗传学。最后，我们将通过神经调节来增强或破坏 LDRN 编码和动机行为的表达。多重 1 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随意增强 LDRN 编码和动机表达这些研究可以共同揭示神经精神疾病的常见情况。以情感和动机失调为特征，并提出特定的神经化学目标治疗干预。