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.

项目总结 怀孕期间过量摄入富含脂肪的饮食会导致后代容易暴饮暴食 变胖并变得肥胖。这种对进食行为的产前编程的世代效应导致了 容易变得肥胖的后代。后代的这种倾向归因于起源的增加 下丘脑中的嗜食性脑啡肽神经元。下丘脑脑啡肽的高水平和高表达 是高脂肪饮食摄入量的刺激物。这种变化的一种潜在机制涉及趋化因子CXCL12， 这一点已被证明会因摄入高脂肪饮食而改变。母体CXCL12，类似于高脂饮食， 还可以刺激脑啡肽水平和后代的摄食行为。目前，尚不清楚如何产前 高脂饮食通过CXCL12影响胚胎发育中的下丘脑结构。 这项建议的目标是在使用大鼠模型的情况下，试验降低母体CXCL12水平是否可以 防止后代大脑发生变化。目的1a将检测CXCL12中和抗体与 母体摄入高脂蛋白对子代下丘脑的影响。利用新的空间基因转录组 神经元和神经胶质细胞的免疫荧光组织学标记技术 将进行起源和食欲神经肽的实验。之后，下丘脑的离散区域将 被条形码编码后，将收集用于RNAseq的mRNA(得到的cDNA)。结果将提供独一无二的 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将测试这些相同的内容 后代的行为和体重变化与肥胖倾向的大鼠一致。每日卡路里摄入量和体重 将跟踪变化，并将进行行为测试，以衡量类似焦虑的行为，这是吞噬过多的标志 接受检查。这些结果将进一步支持HFD诱导母体CXCL12增加的假设 在产生高吞噬后代方面发挥作用，并提供对神经发生的新的更高分辨率的见解 后代大脑中的事件。