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: 10686263
• 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/10686263
• 关键词: Acute; Affect; Appetite Stimulants; Architecture; Bar Codes; Behavioral; Body Weight Changes; Brain; CXCR; CXCR4 gene; Characteristics; Chronic; 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; Stromal Cell-Derived Factor 1; Techniques; Testing; anxiety-like behavior; 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 怀孕期间摄入过量的富含脂肪的饮食会导致后代容易暴饮暴食。 肥胖和变得肥胖。这种出生前摄食行为编程的世代效应导致 容易肥胖的后代。后代的这种倾向性归因于基因的增加， 下丘脑中的食欲脑啡肽神经元。下丘脑脑啡肽的高水平和表达 是高脂肪饮食摄入的刺激剂。这种变化的一个潜在机制涉及趋化因子CXCL 12， 这已经被证明是通过摄入高脂肪饮食改变的。母体CXCL12，类似于高脂肪饮食， 也能刺激后代的脑啡肽水平和摄食行为。目前，尚不清楚产前如何 高脂饮食通过CXCL 12发挥其影响，改变发育中胚胎的下丘脑结构。 该建议使用大鼠模型的目的是试验母体CXCL 12水平的降低是否可以 防止后代大脑发生变化。目的1a将检查CXCL12中和抗体与 母体摄入HFD对后代下丘脑变化的影响。利用新的空间基因转录组 技术，神经元和神经胶质细胞的免疫荧光组织学标记结合 发生和食欲神经肽。之后，下丘脑的离散区域将 将其条形码化，并收集mRNA（所得cDNA）用于RNAseq。结果将提供独特的 基于位置的转录组变化被荧光标记的细胞类型覆盖。目标1b将测试这些相同的 后代的行为和体重变化与肥胖倾向大鼠一致。每日热量摄入和体重 变化将被跟踪，行为测试，以衡量焦虑样行为，一个标志物的暴食症，也将 接受检查。这些结果将进一步支持HFD诱导的母体CXCL 12增加的假设。 在产生吞噬后代中起作用，并为神经发生提供新的更高分辨率的见解 在后代大脑中发生的事件。