Molecular and spatial mapping of hypothalamic cell-types using the Xenium analyzer

使用 Xenium 分析仪绘制下丘脑细胞类型的分子和空间图谱

基本信息

批准号：
10852612
负责人：
Beau Alward
金额：
$ 25万
依托单位：
UNIVERSITY OF HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10852612
关键词：
AR gene Address Aggressive behavior Anatomy Androgen Receptor Androgens Animals Aromatase Atlases Bar Codes Behavior Control Behavioral Brain Brain region Candidate Disease Gene Cell physiology Cells Censuses Cichlids Complex Cues Data Set ENG gene Equipment Estradiol Estrogen Receptors Evolution Exhibits Female Formalin Foundations Freezing Funding Future Gene Conversion Gene Expression Gene Expression Profile Gene Expression Profiling Generations Genes Genetic Genetic Markers Genomics Glucocorticoid Receptor Goals Human Hypothalamic structure In Situ Knowledge Label Laboratories Link Location Maps Molecular Molecular Profiling Mus Neuronal Plasticity Neurophysiology - biologic function Organism Partner in relationship Physiological Population Preoptic Areas Process Progesterone Receptors Research Resolution Role Site Social Behavior Social Controls Social Environment Social Hierarchy Social Interaction Steroids Subcellular Anatomy System Testosterone Tissues Vertebrates Work behavioral response behavioral study cell type cost effectiveness design genetic signature hormonal signals infancy insight interest male mating behavior mind control neuroregulation novel programs response single-cell RNA sequencing social social influence spatial relationship steroid hormone teleost transcriptome whole genome

项目摘要

PROJECT SUMMARY Steroid hormones in humans and other animals coordinate physiological and behavioral processes underlying optimal responses to the social environment. The brain is a major site of steroid hormone action; however, our knowledge of the role of steroid hormones in regulating gene expression and neuroplasticity in the brain is in its infancy. My research program aims to uncover the connections between steroid hormones, gene expression in the brain, and specific cellular functions using Astatotilapia burtoni, a cichlid fish that exhibits sophisticated social hierarchies. In the wild as in the laboratory, male A. burtoni stratify along a social hierarchy where dominant males possess bright coloration, aggressively defend a territory, and mate with females, while non- dominant males do not. Female A. burtoni do not form a social hierarchy but behave aggressively towards one another for mating opportunities. Social rank is in flux, as dominant and non-dominant males can change their status depending on the social milieu. These complex social interactions are tightly linked to levels of a class of steroid hormones called androgens. My research program leverages the social dynamics of A. burtoni in the laboratory to discover the role of androgens in controlling genes in the brain and neuroplasticity. In our ongoing work, we have generated a cell-type specific molecular atlas of the A. burtoni hypothalamus using single-cell RNA-sequencing (scRNAseq). The hypothalamus is a conserved brain region that coordinates adaptive social behaviors across vertebrates. While scRNAseq provides cell-type resolution of specific gene expression patterns, it does not tell you where the specific cells are anatomically within the region of interest. The current proposal aims to address this issue using the 10X Genomics Xenium in situ analyzer, an end-to-end gene expression analysis platform that can label hundreds of genes simultaneously in fresh-frozen or formalin-fixed, sectioned tissue. Using the Xenium will allow for the spatial resolution of the distinct cell-types we have identified in our current scRNAseq dataset. We will combine genetic markers of all cell-types with candidate genes such as distinct androgen receptor (AR) genes present in our Xenium analysis, revealing precisely which cells and where are AR+, providing a precise road-map for testable hypotheses related to the molecular and cellular control of social behavior. Our hypothalamic spatial transcriptome will be compared to the currently only available hypothalamic spatial transcriptome in mice to reveal novel insights into the genetic signatures of AR+ cell types and other cell populations in two species used heavily in social behavior studies. With the generation of a molecular and spatial map of the A. burtoni hypothalamus, we will be able to address fundamental questions regarding the molecular and neural control of the brain and social behavior. Indeed, these questions may connect naturally to those on the control of social behavior in other species such as humans and how social systems emerge throughout evolution.

项目概要人类和其他动物的类固醇激素协调生理和行为过程对社会环境的最佳反应。大脑是类固醇激素作用的主要部位；然而，我们的关于类固醇激素在调节大脑基因表达和神经可塑性中的作用的知识是它的婴儿期。我的研究项目旨在揭示类固醇激素与基因表达之间的联系大脑中的功能，以及使用Astatotilapia burtoni（一种慈鲷鱼，表现出复杂的功能）的特定细胞功能社会等级制度。无论是在野外还是在实验室中，雄性伯托尼 A. burtoni 都按照社会等级分层，其中占主导地位的雄性拥有鲜艳的颜色，积极捍卫领地，并与雌性交配，而非占主导地位的雄性则不然。雌性伯托尼 A. burtoni 不形成社会等级制度，但对社会等级制度表现出攻击性另一个是为了交配机会。社会等级在不断变化，因为主导和非主导的男性可以改变他们的地位地位取决于社会环境。这些复杂的社交互动与一类人的水平紧密相关。类固醇激素称为雄激素。我的研究项目利用了 A. burtoni 的社会动态实验室发现雄激素在控制大脑基因和神经可塑性中的作用。在我们正在进行的工作中，我们使用单细胞生成了 A. burtoni 下丘脑的细胞类型特定分子图谱 RNA 测序 (scRNAseq)。下丘脑是一个保守的大脑区域，负责协调适应性社交脊椎动物的行为。 scRNAseq 提供特定基因表达的细胞类型分辨率模式，它不会告诉您特定细胞在解剖学上位于感兴趣区域内的位置。目前的该提案旨在使用 10X Genomics Xenium 原位分析仪（一种端到端基因）解决此问题表达分析平台，可同时标记新鲜冷冻或福尔马林固定的数百个基因，切片组织。使用 Xenium 将允许我们拥有的不同细胞类型的空间分辨率在我们当前的 scRNAseq 数据集中识别出。我们将把所有细胞类型的遗传标记与候选细胞结合起来我们的 Xenium 分析中存在的基因，例如不同的雄激素受体 (AR) 基因，准确地揭示了哪些细胞以及何处是 AR+，为与分子相关的可测试假设提供精确的路线图以及社会行为的细胞控制。我们的下丘脑空间转录组将与目前的进行比较唯一可用的小鼠下丘脑空间转录组，揭示了对基因特征的新见解两个物种的 AR+ 细胞类型和其他细胞群在社会行为研究中大量使用。随着生成 A. burtoni 下丘脑的分子和空间图，我们将能够解决关于大脑和社会行为的分子和神经控制的基本问题。的确，这些问题可能自然地与其他物种的社会行为控制问题联系起来，例如人类以及社会系统如何在进化过程中出现。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genetic dissection of steroid-hormone modulated social behavior: Novel paralogous genes are a boon for discovery.

类固醇激素调节社会行为的基因剖析：新的旁系同源基因是发现的福音。

DOI：
发表时间：
2023-01
期刊：
影响因子：
0
作者：
Alward, Beau A;Hoadley, Andrew P;Jackson, Lillian R;Lopez, Mariana S
通讯作者：
Lopez, Mariana S

Androgen receptor deficiency is associated with reduced aromatase expression in the ventromedial hypothalamus of male cichlids.

雄激素受体缺乏与雄性慈鲷腹内侧下丘脑芳香酶表达减少有关。

DOI：
发表时间：
2024-02
期刊：
影响因子：
5.2
作者：
Lopez, Mariana S;Alward, Beau A
通讯作者：
Alward, Beau A

Sex differences in aggression and its neural substrate in a cichlid fish.

慈鲷攻击性的性别差异及其神经基质。

DOI：
发表时间：
2024-02-10
期刊：
影响因子：
0
作者：
Jackson, Lillian R;Dumitrascu, Mariam;Alward, Beau A
通讯作者：
Alward, Beau A

Female cichlids attack and avoid-but will still mate with-androgen receptor mutant males that lack male-typical body coloration.

雌性丽鱼会攻击并避开缺乏雄性典型体色的雄激素受体突变雄性，但仍会与之交配。

DOI：
发表时间：
2023-11-04
期刊：
影响因子：
0
作者：
Howard, Megan R;Ramsaroop, Maxximus G;Hoadley, Andrew P;Jackson, Lillian R;Lopez, Mariana S;Saenz, Lauren A;Alward, Beau
通讯作者：
Alward, Beau

Breaking Through the Bottleneck: Krogh's Principle in Behavioral Neuroendocrinology and the Potential of Gene Editing.

突破瓶颈：克罗行为神经内分泌学原理和基因编辑的潜力。