Activity-dependent Regulation of the Choroid Plexus-Cerebrospinal Fluid Stem Cell Niche

脉络丛-脑脊液干细胞生态位的活动依赖性调节

• 批准号: 10626875
• 负责人: MARIA LEHTINEN
• 金额: $ 64.31万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10626875
• 关键词: Affect; Agonist; Apical; Axon; Bathing; Brain; Brain Stem; Calcium; Cells; Cerebral cortex; Cerebrospinal Fluid; Choroid Plexus Epithelium; Couples; Cues; Data; Development; Diffusion; Embryo; Embryonic Development; Environment; Epithelial Cells; Epithelium; Fetal health; Future; G-Protein-Coupled Receptors; Gene Expression; Gene Proteins; Genetic; Genetic Transcription; Growth; Health; Image; Immediate-Early Genes; In Vitro; Intraperitoneal Injections; Investigation; Ligands; Mediating; Methods; Modernization; Molecular; Monitor; Mus; Nature; Neurodevelopmental Disorder; Neurologic; Neurons; Penetrance; Peripheral; Phenotype; Placenta; Process; Production; Protein Secretion; Receptor Activation; Receptor Signaling; Regulation; Salivary Glands; Serotonin; Signal Pathway; Signal Transduction; Source; Stimulus; Structure of choroid plexus; Surface; Testing; brain tissue; direct application; experimental study; fetal; gene induction; in vivo; in vivo calcium imaging; intraperitoneal; loss of function; nerve stem cell; neurosteroids; novel; postnatal; receptor; response; stem cell niche; tool; transcriptome; transcriptome sequencing; transcriptomics; vesicular release

PROJECT SUMMARY / ABSTRACT Building the cerebral cortex requires the spatial and temporal orchestration of molecular signals across neural progenitor cells. Each of these cells is bathed in cerebrospinal fluid (CSF), a medium rich in health- and growth- promoting factors whose composition changes profoundly across days during brain development. The choroid plexus (ChP) is likely to strongly contribute to this dynamic developmental regulation of CSF composition, but the underlying mechanisms are not understood. Here, we will use a suite of modern tools to test the hypothesis that elevations in calcium concentration in embryonic ChP epithelial cells in response to extrinsic cues is critical for regulating synthesis and secretion of signaling factors into the CSF. Calcium activity regulates transcription and secretion in diverse cells ranging from neurons to salivary gland epithelia. We identified receptors expressed in ChP epithelial cells that mediate calcium entry into the cell, such as the TRPM3 receptor, or that drive calcium release from internal stores, such as the 5HT2C receptor (Htr2c) for serotonin (5- HT, 5-hydroxytryptamine). Our preliminary data suggest that 5-HT regulates embryonic ChP calcium activity, transcription, and vesicle release, and that intact signaling via the 5HT2C receptor in ChP is essential for normal cerebral cortical development. Further, maternal delivery of a selective agonist for the 5HT2C receptor triggers rapid, robust immediate early gene expression in embryonic ChP. Thus, we propose that ChP calcium activity triggered by signals that activate the 5HT2C receptor (and other receptors such as TRPM3) regulates ChP transcription and secretion of signaling factors into the CSF and controls body-brain and maternal-fetal interactions, thereby tightly orchestrating cerebral cortical development. To test this, we will first investigate the effects of 5HT2C and TRPM3 receptor activation by direct application of ligand to the apical surface of epithelial cells during calcium imaging in ChP explants (Aim 1a) and by stimulation of the basal surface following intraperitoneal (IP) delivery of ligands to the maternal dam during in vivo two- photon calcium imaging of attached embryos (Aim 1b). We then assess the transcriptomic impact of such stimulation (Aim 1c), and test whether embryonic ChP calcium activity via activation of 5HT2C receptors and other receptors triggers vesicle release and protein secretion in vitro (Aim 2a) and in vivo (Aim 2b). We will also analyze the effects of disruption of ChP receptor signaling on cortical development (Aim 3a). Disrupted CSF volume, composition, and ventricle formation are common to many neurological conditions. Perturbations in maternal-fetal health can disrupt brain development, in part via dysregulation of serotonergic and other signaling pathways. Our findings provide a novel platform for elucidating the mechanisms underlying dysregulation of brain development by environmental perturbations, thereby providing a conceptual and technical roadmap for future studies on how activity in ChP epithelial cells influences gene expression and protein secretion, with consequences for many neurodevelopmental diseases.

项目摘要 /摘要 建立大脑皮层需要分子信号跨神经的空间和时间编排 祖细胞。这些细胞中的每一个都沐浴在脑脊液（CSF）中，这是一种富含健康和生长的培养基 促进其组成在大脑发育过程中在整个时期发生深刻变化的因素。脉络膜 丛（CHP）可能对CSF组成的这种动态发育调节有很大贡献，但是 基本机制尚不清楚。在这里，我们将使用一套现代工具来测试 假设胚胎CHP上皮细胞中钙浓度的升高是响应于 外在提示对于调节CSF的信号传导因子的合成和分泌至关重要。钙 活性调节从神经元到唾液腺上皮的不同细胞的转录和分泌。我们 在CHP上皮细胞中鉴定的受体介导钙进入细胞，例如TRPM3 受体，或从内部商店驱动钙释放的钙，例如5-羟色胺的5HT2C受体（HTR2C）（5-- HT，5-羟色胺）。我们的初步数据表明5-HT调节胚胎CHP钙 活性，转录和囊泡释放，以及通过CHP中5HT2C受体的完整信号传导为 对于正常的脑皮质发育至关重要。此外，孕产妇的选择性激动剂 5HT2C受体在胚胎CHP中触发快速，可靠的早期基因表达。因此，我们建议 通过激活5HT2C受体的信号触发的CHP钙活性（以及其他受体此类受体 作为TRPM3）调节CHP转录和信号传导因子的分泌到CSF和对照中 身体脑和孕产妇的相互作用，从而紧密地精心策划了脑皮质发育。 为了测试这一点，我们将首先通过直接应用直接应用5HT2C和TRPM3受体激活的影响 在CHP外植体中钙成像期间（AIM 1A）和通过刺激 腹膜腹膜（IP）在体内两种体内递送到母体大坝后的基底表面 附着的胚胎的光子钙成像（AIM 1B）。然后，我们评估此类的转录组影响 刺激（AIM 1C），并通过5HT2C受体的激活来测试胚胎CHP钙的活性 其他受体会在体外触发囊泡释放和蛋白质分泌（AIM 2A）和体内（AIM 2B）。我们也会 分析CHP受体信号破坏对皮质发育的影响（AIM 3A）。 CSF中断 体积，成分和心室形成是许多神经系统条件共有的。扰动 孕产妇健康会破坏大脑发育，部分通过血清素能和其他信号传导失调 途径。我们的发现为阐明大脑失调的机制提供了一个新的平台 通过环境扰动开发，从而为未来提供概念和技术路线图 CHP上皮细胞中的活性如何影响基因表达和蛋白质分泌的研究， 许多神经发育疾病的后果。

项目成果

Cerebrospinal Fluid Magnetic Resonance Imaging: Improving Early Diagnosis of Autism and Other Neurodevelopmental Conditions.

脑脊液磁共振成像：改善自闭症和其他神经发育疾病的早期诊断。

• DOI: 10.1016/j.bpsc.2020.05.007
• 发表时间: 2020
• 期刊: Biological psychiatry. Cognitive neuroscience and neuroimaging
• 作者: Xu,Huixin;Lehtinen,MariaK
• 通讯作者: Lehtinen,MariaK

Development and functions of the choroid plexus-cerebrospinal fluid system.

• DOI: 10.1038/nrn3921
• 发表时间: 2015-08
• 期刊: Nature reviews. Neuroscience
• 作者: Lun MP;Monuki ES;Lehtinen MK
• 通讯作者: Lehtinen MK

Progressive Differentiation and Instructive Capacities of Amniotic Fluid and Cerebrospinal Fluid Proteomes following Neural Tube Closure.

• DOI: 10.1016/j.devcel.2015.11.015
• 发表时间: 2015-12-21
• 期刊: Developmental cell
• 影响因子: 11.8
• 作者: Chau KF;Springel MW;Broadbelt KG;Park HY;Topal S;Lun MP;Mullan H;Maynard T;Steen H;LaMantia AS;Lehtinen MK
• 通讯作者: Lehtinen MK

Disruption of GMNC-MCIDAS multiciliogenesis program is critical in choroid plexus carcinoma development.

• DOI: 10.1038/s41418-022-00950-z
• 发表时间: 2022-08
• 期刊: Cell death and differentiation
• 影响因子: 12.4
• 作者: Li Q;Han Z;Singh N;Terré B;Fame RM;Arif U;Page TD;Zahran T;Abdeltawab A;Huang Y;Cao P;Wang J;Lu H;Lidov HGW;Surendran K;Wu L;Virga JQ;Zhao YT;Schüller U;Wechsler-Reya RJ;Lehtinen MK;Roy S;Liu Z;Stracker TH;Zhao H
• 通讯作者: Zhao H

Antibodies Expand the Scope of Angiotensin Receptor Pharmacology.

抗体扩大了血管紧张素受体药理学的范围。

• DOI: 10.1101/2023.08.23.554128
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Skiba,MeredithA;Sterling,SarahM;Rawson,Shaun;Gilman,MorganSA;Xu,Huixin;Nemeth,GenevieveR;Hurley,JosephD;Shen,Pengxiang;Staus,DeanP;Kim,Jihee;McMahon,Conor;Lehtinen,MariaK;Wingler,LauraM;Kruse,AndrewC
• 通讯作者: Kruse,AndrewC