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

• 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
• 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

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.

项目总结 人类和其他动物中的类固醇激素协调潜在的生理和行为过程 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 关于类固醇激素在大脑中调节基因表达和神经可塑性的作用的知识在 它还处于初级阶段。我的研究计划旨在揭示类固醇激素、基因表达 在大脑中，以及特定的细胞功能中使用了伯顿拟罗非鱼，这是一种显示出复杂的鱼纲鱼类 社会等级制度。在野外，就像在实验室里一样，雄性伯通尼亚种沿着社会等级分层，在那里 占优势的雄性拥有鲜艳的色彩，咄咄逼人地保卫领土，并与雌性交配，而非 占统治地位的雄性则不会。雌性A.burtoni不会形成一个社会等级，但会对一个等级表现出攻击性 另一个是交配机会。社会地位正在变化，因为占主导地位和非占主导地位的男性可以改变他们的 地位取决于社会环境。这些复杂的社会互动与一类 被称为雄激素的类固醇激素。我的研究项目利用了A.burtoni在 实验室发现雄激素在控制大脑中的基因和神经可塑性方面的作用。在我们正在进行的 在这项工作中，我们已经用单细胞构建了伯顿沙门氏菌下丘脑的细胞型特异性分子图谱。 RNA测序(ScRNAseq)。下丘脑是一个保守的大脑区域，负责协调适应性社会 脊椎动物的行为。而scRNAseq提供特定基因表达的细胞类型解析 模式，它不会告诉你特定的细胞在感兴趣的区域内的解剖位置。海流 该提案旨在使用端到端基因的10倍基因组Xum原位分析仪来解决这一问题 可以同时标记数百个新鲜冷冻或福尔马林固定的基因的表达分析平台， 切片的组织。使用Xum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。我们将结合所有细胞类型的遗传标记和候选 我们的Xum分析中存在不同的雄激素受体(AR)基因等基因，准确地揭示了 AR+是哪些细胞以及在哪里，为与分子相关的可测试假说提供了精确的路线图 以及对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 只有可用的小鼠下丘脑空间转录组才能揭示对人类免疫缺陷病毒基因特征的新见解 AR+细胞类型和其他细胞群在两个物种中被大量用于社会行为研究。与 伯顿支原体下丘脑的分子和空间图谱的生成，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， 这些问题可能自然而然地与其他物种对社会行为的控制有关，例如 人类和社会制度如何在整个进化过程中出现。

项目成果

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

• DOI: 10.1016/j.yhbeh.2022.105295
• 发表时间: 2022-12
• 期刊: Hormones and Behavior
• 影响因子: 3.5
• 作者: Beau A. Alward;Andrew P. Hoadley;Lillian R. Jackson;Mariana S. Lopez

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: 10.1101/2023.11.02.565323
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Howard,MeganR;Ramsaroop,MaxximusG;Hoadley,AndrewP;Jackson,LillianR;Lopez,MarianaS;Saenz,LaurenA;Alward,Beau
• 通讯作者: Alward,Beau

Breaking through the bottleneck: Krogh's principle in behavioral neuroendocrinology and the potential of gene editing.

• DOI: 10.1093/icb/icad068
• 发表时间: 2023-06
• 期刊: Integrative and comparative biology
• 影响因子: 2.6
• 作者: Lillian R. Jackson;Mariana S. Lopez;Beau A. Alward