Cannabinoid Signaling Interactions During Axon Development in situ

轴突原位发育过程中大麻素信号相互作用

基本信息

批准号：
10654243
负责人：
Tamira Elul
金额：
$ 36.43万
依托单位：
TOURO UNIVERSITY OF CALIFORNIA
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：
2023-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10654243
关键词：
Actins Adhesives Affect Age Animal Model Axon Behavioral Binding Biological Biological Models Brain Breast Cancer Cell CNR1 gene Cadherins California Cannabinoids Cannabis Cell physiology Cell-Cell Adhesion Cells Child Clinical Cognitive Cognitive deficits Collaborations Communication Complex Couples Critical Thinking Cyclic AMP Cyclic AMP-Dependent Protein Kinases Data Data Analyses Defect Development Dominant-Negative Mutation Endocannabinoids Environment Evidence Based Medicine Evidence based practice Exposure to Extramural Activities Faculty Fetal Development Fetus Filopodia Fishes Funding G-Protein-Coupled Receptors Genetic Goals Grant Growth Cones Health Professional Human Image In Situ In Vitro Individual Journals Knowledge Laboratories Laboratory Research Lead Literature Marijuana Mediating Medical Students Modeling Molecular Molecular Biology Morphology Mus Muscle fasciculation Neurobiology Neurons Outcome Paper Patient Care Phenotype Plants Pregnant Women Process Publications Publishing Quantitative Microscopy Rana Regulation Research Retinal Ganglion Cells Schools Signal Pathway Signal Transduction Structure Students Substance of Abuse System Tadpoles Testing Time Universities Vertebrates WNT Signaling Pathway Woman Work Xenopus Xenopus laevis axon growth axonal pathfinding cancer cell cannabinoid receptor career experience fetal fetal marijuana exposure genetic manipulation hashish improved in vivo loss of function marijuana use matriculation meetings mutant netrin receptor neurobehavioral neuron development neuronal circuitry novel pharmacologic posters prenatal prenatal exposure reproductive retinotectal rho skills stem substance use vision development

项目摘要

Abstract The cannabis plants marijuana and hashish are the most commonly used substances of abuse by women of reproductive age. But, concerningly, frequent use of cannabis by pregnant women may result in lasting cognitive and neuro-behavioral issues in children that were exposed in utero. The mechanisms by which cannabinoids in cannabis plants influence the structure and function of the developing brain are not well understood. Previous studies in animal models indicate that the main cannabinoid receptor, CB1R, affects formation of neuronal circuits by signaling though various molecular factors such as the Netrin receptor DCC, PKA and RhoA. However, we lack comprehensive knowledge of essential mechanisms by which CB1R influences neuronal circuit formation during brain development. For many years, our laboratory determined signaling mechanisms for Wnt and Cadherin factor, -catenin, in development of the visual projection in tadpoles of the vertebrate frog model Xenopus laevis. This is an ideal system for studying neuronal circuit formation because of its strong genetic similarity to humans, amenability to molecular and genetic manipulation in single retinal ganglion cells and imaging of individual retinal ganglion cells with altered molecular signaling directly in their native environment. We now propose to determine whether CB1R inhibits Wnt/Cadherin/- catenin signaling to regulate growth cone filopodia and axon pathfinding features in situ. This proposal stems from work from others showing that CB1R inhibits Wnt and destabilizes -catenin in cancer cells, and Wnt mediated destabilization of -catenin downregulates Cadherin cell-cell adhesion, and our recently published paper showing that CB1R and -catenin oppositely modulate growth cone filopodia in retinal ganglion cells. We will test two aims: 1) Quantify axon pathfinding and growth cone defects following manipulation of CB1R in situ. Our previously published data showed that pharmacological manipulation of CB1R perturbs growth cone filopodia and axon fasciculation in situ. We will now assess whether cell- autonomous (morpholino based) loss-of-function of CB1R in individual retinal ganglion cells alters multiple growth cone and axon pathfinding parameters. 2) Establish functional interactions between CB1R and Wnt/Cadherin signaling in retinal ganglion cells. We will determine whether phenotypic effects of CB1R loss- of-function on growth cone filopodia and axon pathfinding features are rescued by expression of factors in the canonical Wnt signaling pathway (Axin, APC) and mutants of key players in Cadherin cell-cell adhesive complex (-catenin, -catenin). This data will determine a novel and essential signaling mechanism for cannabinoids in neuronal circuit development in a vertebrate model. Given the conservation of these signaling pathways, these results may also establish a fundamental mechanism for endocannabinoids in formation of neuronal circuits in human fetuses, and for how increased prenatal cannabis exposure could disrupt establishment of neuronal circuits, and lead to persistent cognitive and neurobehavioral deficits in children.

抽象的大麻植物大麻和大麻是妇女最常用的虐待物质生殖年龄。但是，关于孕妇经常使用大麻可能会导致持久在子宫内暴露的儿童的认知和神经行为问题。所在的机制大麻植物中的大麻素会影响发育大脑的结构和功能不好理解。动物模型的先前研究表明，主要大麻素受体CB1R会影响通过信号传导各种分子因子（例如Netrin受体DCC）来形成神经元回路 PKA和Rhoa。但是，我们缺乏对CB1R的基本机制的全面知识影响大脑发育过程中神经元电路的形成。多年以来，我们的实验室确定 Wnt和cadherin因子（-catenin）的信号传导机制在开发视觉投影中脊椎动物青蛙模型Laevis的t。这是研究神经元电路的理想系统由于其与人类的遗传相似性很强，因此对分子和遗传操纵的遗传性在单个视网膜神经节细胞和具有改变分子信号的单个视网膜神经节细胞的成像中直接在其本地环境中。现在，我们建议确定CB1R是否抑制Wnt/Cadherin/-- Catenin信号传导以原位调节生长锥丝状和轴突探路特征。这个建议来自其他工作的原因，表明CB1R抑制Wnt并破坏癌细胞中的-catenin，并且 Wnt介导的-catenin介导的不稳定下调Cadherin细胞 - 细胞粘合剂，而我们最近发表的论文表明，CB1R和-catenin在视网膜中反对调节生长锥神经节细胞。我们将测试两个目的：1）量化轴突路径和生长锥缺陷。在原位操纵CB1R。我们先前发布的数据表明，药物操纵 CB1R将生长锥丝状和轴突诱发。现在，我们将评估细胞是否是否 CB1R在单个残留神经节细胞中的自主（基于形态的）功能丧失会改变多个生长锥和轴突探路参数。 2）在CB1R和视网膜神经节细胞中的Wnt/cadherin信号传导。我们将确定CB1R损失的表型效应是否生长锥丝状和轴突探索特征的功能是由因子表达的反应 CADHERIN细胞 - 细胞粘合剂中主要参与者的Canonical Wnt信号通路（AXIN，APC）和突变体复合物（-catenin，-catenin）。这些数据将确定一种新颖的基本信号传导机制脊椎动物模型中神经元电路发育中的大麻素。考虑到这些信号的保护途径，这些结果还可能建立一种基本机制，以形成内源性大麻素人类胎儿的神经元电路，以及增加产前大麻的暴露可能会破坏建立神经元电路，并导致儿童持续的认知和神经行为缺陷。