The role of Tex15 in shaping stochastic olfactory receptor gene choice.

Tex15 在塑造随机嗅觉受体基因选择中的作用。

基本信息

批准号：
10826734
负责人：
Nusrath Yusuf
金额：
$ 3.91万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
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：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10826734
关键词：
Adolescent Alleles Binding Binding Proteins Biochemical ChIP-seq Chemicals Chromatin Chromatin Structure Complex DNA Methylation DNA Transposable Elements DNA methylation profiling Data Deposition Detection Development Down-Regulation Elements Ensure Environment Epigenetic Process Excision Faculty Fellowship Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Transcription Genomics Heterochromatin Histones KDM1A gene Knock-out Knockout Mice Mammals Maps Mediating Mentorship Methylation Molecular Mus Mutant Strains Mice Nervous System Neuronal Differentiation Nuclear Odorant Receptors Olfaction Disorders Olfactory Epithelium Pattern Perception Phenotype Process Proteins Receptor Gene Regulator Genes Repression Rest Role Sensory Shapes Smell Perception Specificity Spermatocytes Students Techniques Testing Testis Training Transcriptional Silencer Elements Up-Regulation Work chemical binding computational neuroscience experimental study gene repression genome-wide histone demethylase histone methyltransferase innovation insight link protein methylation pattern mutant nerve stem cell novel olfactory receptor olfactory sensory neurons piRNA programs promoter receptor expression recruit single-cell RNA sequencing transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT The incredible specificity with which we can discern a vast diversity of smells emerges from how olfactory sensory neurons (OSN) express olfactory receptor (OR) genes. These OR genes encode the proteins that bind chemical odorants. Each OSN stochastically transcribes only one allele of an OR gene. An OSN can choose to express any one of the approximately 1400 OR genes or 2800 alleles. There is no one to one relationship between an odorant and an OR as a combination of ORs are involved in the detection of one odorant. Thus, the diversity of ORs expressed is important to the main olfactory epithelium’s (MOE) ability to detect a vast range of smells. The choice of an OR gene occurs as OSN progenitors mature into an OSN. Recent work has revealed remarkable remodeling of the local chromatin and co-expression of multiple OR genes during this period, but the molecular mechanisms governing these processes and their connection to a singular OR allele expression in a mature OSN remain unknown. We show that testis expressed 15 (Tex15), a protein that has only been studied in the testes where it regulates methylation and silencing of transposons, is crucial for stochastic OR gene choice. We find that when Tex15 is knocked out there is a dramatic reduction in the diversity of expressed OR genes with a few OR genes dominating stochastic choice. Strikingly, Tex15 is only transiently expressed in OSN progenitors during a period of OR co-expression. Our central hypothesis is that Tex15 is epigenetically controlling stochastic OR gene choice, either by regulating DNA methylation or other aspects of OR gene chromatin structure. Aim 1 will determine whether Tex15 is required for transient co-expression of multiple OR genes in OSN progenitors. In aim 2, we will determine whether Tex15 protein binds to OR genes. Finally in aim 3, our experiments on primary OSNs from Tex15 null mice will determine how gene repression is mediated in OSN progenitors – through methylation or deposition of repressive heterochromatin. Together, these experiments will elucidate a novel aspect of how OSNs come to stochastically choose a single OR and how Tex15 gene and protein guides this specific yet diverse choice. We anticipate this project will uncover new and/or unexpected role for Tex15 in the MOE. This will also reveal the key molecular underpinnings for how our sense of smell develops and may provide important insight to further understand the mechanisms of smell disorder. The proposed fellowship will train this applicant in several innovative techniques in biochemical, cellular, molecular, and computational neuroscience. The established faculty-student mentorships will ensure proper scientific and professional training necessary to become a successful, independent sensory and computational neurobiologist.

项目概要/摘要我们能够辨别出多种多样的气味，其令人难以置信的特异性来自于嗅觉如何感觉神经元 (OSN) 表达嗅觉受体 (OR) 基因，这些 OR 基因编码结合的蛋白质。每个 OSN 随机转录一个 OR 基因的一个等位基因。表达大约 1400 个 OR 基因或 2800 个等位基因中的任何一个不存在一对一的关系。气味剂和 OR 之间的关系作为 OR 的组合参与一种气味剂的检测。表达的 OR 的多样性对于主嗅觉上皮 (MOE) 检测大量信息的能力非常重要气味范围。最近的研究表明，OR 基因的选择发生在 OSN 祖细胞成熟为 OSN 的过程中。在此期间，局部染色质发生了显着的重塑，并且多个 OR 基因共表达，但是控制这些过程的分子机制及其与单一 OR 等位基因表达的联系成熟的 OSN 中的情况仍然未知。我们发现睾丸表达 15 (Tex15)，这种蛋白质仅在睾丸中进行过研究，它调节着转座子的甲基化和沉默对于随机 OR 基因选择至关重要。敲除少数 OR 基因后，表达的 OR 基因的多样性急剧下降引人注目的是，Tex15 仅在 OSN 祖细胞中短暂表达。我们的中心假设是 Tex15 在表观遗传上控制随机 OR。基因选择，要么通过调节 DNA 甲基化，要么通过基因染色质结构的其他方面进行选择。确定 OSN 中多个 OR 基因的瞬时共表达是否需要 Tex15 在目标 2 中，我们将确定 Tex15 蛋白是否与 OR 基因结合。对 Tex15 缺失小鼠的原代 OSN 进行的实验将确定基因抑制是如何介导的 OSN 祖细胞——通过甲基化或抑制性异染色质沉积。总之，这些实验将阐明 OSN 如何随机选择单个节点的新颖方面或者，以及 Tex15 基因和蛋白质如何指导这一特定但多样化的选择，我们预计该项目将会。揭示 Tex15 在 MOE 中新的和/或意想不到的作用这也将揭示关键分子。我们的嗅觉如何发展的基础，并可能为进一步理解嗅觉提供重要的见解拟议的奖学金将为该申请人提供几种创新技术的培训。生物化学、细胞、分子和计算神经科学领域的知名师生。指导将确保成为成功的、独立的感觉和计算神经生物学家。