Transcription Factors Governing the Development of GHRH-neurons

控制 GHRH 神经元发育的转录因子

• 批准号: 10676744
• 负责人: JAE W LEE
• 金额: $ 54.79万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676744
• 关键词: Area; Binding; Biochemical; Cells; ChIP-seq; Data; Data Set; Development; Developmental Gene; Dwarfism; Embryo; Embryonic Development; Ensure; Exclusion; FOXP2 gene; Genes; Genome Mappings; Goals; Grant; Growth; Growth and Development function; Homeostasis; Hypothalamic structure; Immunohistochemistry; Impairment; In Situ Hybridization; Individual; Knockout Mice; Life; Link; Metabolic Diseases; Methods; Molecular; Mus; Mutant Strains Mice; Neurons; Pathway interactions; Pattern; Physiological; Play; Process; Production; Regulation; Regulator Genes; Regulatory Pathway; Reporting; Reproduction; Research; Role; Somatotropin-Releasing Hormone; Structure of nucleus infundibularis hypothalami; Testing; Time; Work; biomarker panel; cell type; chromatin immunoprecipitation; conditional knockout; course development; energy balance; experimental study; feeding; fighting; gene regulatory network; genome-wide; genome-wide analysis; improved; in vivo; innovation; insight; mouse genetics; mouse genome; neuron development; novel; postnatal; progenitor; programs; single-cell RNA sequencing; transcription factor; transcriptome

The ultimate research goal of this lab is to decipher the gene regulatory network that directs the development of various types of neurons in the mouse arcuate nucleus of the hypothalamus (ARC). Despite the physiological significance of many ARC neurons, the developmental gene regulatory programs for ARC neurons remain poorly understood. This lab has been pioneering this emerging area of studies by successfully combining mouse genetics and genome-wide studies. Notably, a close developmental link has been discovered among different types of ARC neurons. These intertwined developmental pathways are likely crucial to ensure the balanced production of different ARC neurons during embryogenesis, enabling a highly coordinated regulation of various homeostatic processes in later postnatal life, such as integration of feeding, reproduction, and growth. Key preliminary results in this grant include: i) Single cell RNA-seq (scRNA-seq) analyses reveal eight TFs enriched in developing growth hormone-releasing hormone (GHRH)-neurons in the ARC, which control linear growth; Prox1, Gsx1, Egr1, Foxp2, Pbx3, St18, Dlx1 and Dlx2. Notably, Dlx1/2, Foxp2 and Gsx1 have been shown to be important for the development of GHRH-neurons, indicating that the scRNA-seq approach is highly useful to identify TFs acting on neuronal lineage development. ii) GHRH-specific inactivation of Prox1 leads to dwarfism and reduced Ghrh expression in mice. iii) Further, ChIP-seq data for Dlx1 provides various new insights into the mechanism by which Dlx1 controls GHRH-neuronal development. Together, these results led to the central hypothesis Dlx1/2 and Prox1 play vital roles in acquiring GHRH-neuronal fate over other related ARC neuronal lineages in part by coordinating the expression of downstream TFs. This hypothesis will be tested in the two specific aims using an ensemble of biochemical and cellular methods, mouse genetics and genome-wide approaches. Completion of this innovative study will radically improve the understanding of how common progenitors are guided to gain a specific ARC lineage identity over other related cell fates, providing a critical mechanism contributing to the balanced production of diverse ARC neuronal types during development.

该实验室的最终研究目标是破译基因调控网络，指导发展， 小鼠下丘脑弓状核（ARC）中的各种神经元。尽管生理上 尽管许多ARC神经元的重要性，ARC神经元的发育基因调控程序仍然很差 明白这个实验室一直是这一新兴研究领域的先驱， 遗传学和全基因组研究。值得注意的是，在不同的人之间发现了一种密切的发展联系， ARC神经元的类型。这些相互交织的发展途径可能对确保平衡发展至关重要。 在胚胎发生过程中产生不同的ARC神经元，使各种神经元的高度协调调节成为可能。 出生后后期的稳态过程，如摄食、生殖和生长的整合。 该资助的主要初步结果包括：i）单细胞RNA-seq（scRNA-seq）分析揭示了八个TF 富含ARC中发育中的生长激素释放激素（GHRH）神经元，其控制线性 生长; Prox 1、Gsx 1、Egr 1、Foxp 2、Pbx 3、St 18、Dlx 1和Dlx 2。值得注意的是，Dlx 1/2、Foxp 2和Gsx 1已经被发现。 显示对于GHRH神经元的发育是重要的，表明scRNA-seq方法是高度有效的。 用于鉴定作用于神经元谱系发育的TF。ii）Prox 1的GHRH特异性失活导致 侏儒症和减少的Ghrh表达。iii）此外，Dlx 1的ChIP-seq数据提供了各种新的见解 Dlx 1控制GHRH神经元发育的机制。总之，这些结果导致了 中心假说Dlx 1/2和Prox 1在获得GHRH-神经元的命运中起着至关重要的作用， ARC神经元谱系部分通过协调下游TF的表达。这一假设将是 使用生物化学和细胞方法，小鼠遗传学和 全基因组方法。这项创新研究的完成将从根本上提高对如何 共同的祖细胞被引导获得超过其他相关细胞命运的特定ARC谱系身份， 在发育过程中平衡产生不同ARC神经元类型的关键机制。