Resolving the effects of dietary fat induced maternal CXCL12 on offspring hypothalamus using spatial gene transcriptomics

利用空间基因转录组学解析膳食脂肪诱导的母体 CXCL12 对后代下丘脑的影响

• 批准号: 10509951
• 负责人: KINNING POON
• 金额: $ 7.99万
• 依托单位: COLLEGE AT OLD WESTBURY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-18 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10509951
• 关键词: Acute; Affect; Appetite Stimulants; Architecture; Bar Codes; Behavioral; Body Weight Changes; Brain; CXCL12 gene; CXCR4 gene; Characteristics; Chronic; Cohort Effect; Complementary DNA; Complex; Detection; Development; Diet; Dietary Fats; Eating; Embryo; Embryonic Development; Energy Intake; Enkephalins; Environment; Event; Exposure to; Fatty acid glycerol esters; Feeding behaviors; Female; Gene Expression; Genes; Goals; High Fat Diet; Histology; Hyperphagia; Hypothalamic structure; Immunofluorescence Immunologic; Inflammation; Inflammation Mediators; Inflammatory; Ingestion; Intake; Label; Location; Measures; Messenger RNA; Modeling; Modification; Motor Activity; Neuroglia; Neurologic; Neurons; Neuropeptides; Obesity; Outcome; Peptides; Physiological; Play; Pregnancy; Process; Proteins; Rattus; Research; Resolution; Risk; Role; Techniques; Testing; anxiety-like behavior; base; behavior change; behavior test; cell type; chemokine; cytokine; dietary excess; in utero; insight; inter-alpha-inhibitor; male; neurogenesis; neuron development; neutralizing antibody; novel; offspring; paraventricular nucleus; prenatal; prevent; receptor; sex; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Excessive intake of a diet rich in fats during pregnancy gives rise to offspring that are prone to overeating dietary fat and becoming obese. This generational effect of prenatal programming of ingestive behavior results in offspring that are prone to becoming obese. This proneness in offspring is attributed to an increase in the genesis of orexigenic enkephalin neurons in the hypothalamus. High levels and expression of hypothalamic enkephalin is a stimulator of high-fat diet intake. One potential mechanism for this change involves the chemokine CXCL12, which has been shown to be altered by the intake of a high-fat diet. Maternal CXCL12, similar to a high-fat diet, can also stimulate enkephalin levels and ingestive behavior in offspring. Currently, it is unknown how prenatal high-fat diet exerts its affects through CXCL12 to change the hypothalamic architecture in developing embryos. The goal of this proposal in using a rat model is pilot whether the reduction of maternal CXCL12 levels could prevent offspring brain changes. Aim 1a will examine the effects of CXCL12-neutralizing antibody paired with maternal HFD ingestion on changes in offspring hypothalamus. Utilizing the novel spatial gene transcriptome technique, immunofluorescence histology labeling of neurons and neuroglia in conjunction with markers of genesis and orexigenic neuropeptides will be performed. Afterwards, discrete regions of the hypothalamus will be barcoded, and the mRNA (resulting cDNA) will be collected for RNAseq. The results will provide unique location-based transcriptome changes overlaid by fluorescently labeled cell types. Aim 1b will test these same offspring for behavioral and weight changes that align with obesity-prone rats. Daily caloric intake and weight change will be tracked and behavioral tests to measure anxiety-like behavior, a marker for hyperphagia, will also be examined. These results will further support the hypothesis that HFD-induced increase in maternal CXCL12 plays a role in producing hyperphagic offspring and provide new higher resolution insight into neurogenesis events in offspring brain.

项目摘要 怀孕期间富含脂肪的饮食过多的摄入量会引起后代，这些后代容易暴饮暴食 脂肪和肥胖。产前摄入行为的产前编程的这种代效果导致 容易肥胖的后代。后代中的这种pre骨归因于创世记的增加 下丘脑中的甲虫enkephalin神经元。下丘脑的高水平和表达 是高脂饮食摄入量的刺激剂。这种变化的一种潜在机制涉及趋化因子CXCL12， 这已显示出高脂饮食的摄入量改变。母体CXCL12，类似于高脂饮食， 还可以刺激后代中的脑链酸水平和摄取行为。目前，尚不清楚产前 高脂饮食通过CXCL12施加影响，以改变发展胚胎中的下丘脑结构。 使用大鼠模型的该建议的目的是试点母体CXCL12水平的降低是否可以 防止后代大脑的变化。 AIM 1A将检查CXCL12中和抗体的影响 母体HFD摄入后代下丘脑变化。利用新型空间基因转录组 技术，神经元和神经元的免疫荧光组织学标记与标记 将执行创世纪和神经肽。之后，下丘脑的离散区域将 进行条形码，并将收集mRNA（由此产生的cDNA）用于RNASEQ。结果将提供独特的 基于位置的转录组的变化被荧光标记的细胞类型覆盖。 AIM 1B将测试这些 行为和体重变化的后代与肥胖易发的大鼠保持一致。每日热量摄入和体重 将跟踪变化，并进行行为测试以测量焦虑症行为，这是多晶状体的标志物，也将 被检查。这些结果将进一步支持以下假设：HFD诱导的母体CXCL12的增加 在产生倍晶后代中发挥作用，并为神经发生提供新的更高分辨率洞察力 后代大脑中的事件。