Lipidomics analysis of identified single neurons in the adult rodent brain

成年啮齿动物大脑中已识别的单个神经元的脂质组学分析

基本信息

  • 批准号:
    9488668
  • 负责人:
  • 金额:
    $ 0.85万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2016
  • 资助国家:
    美国
  • 起止时间:
    2016-04-01 至 2018-03-31
  • 项目状态:
    已结题

项目摘要

 DESCRIPTION (provided by applicant): Lipids play key roles in brain function and contribute in important ways to pathologies such as drug addiction, schizophrenia and Alzheimer's disease. In preliminary experiments, we utilized a pipette capture method originally devised for single-neuron mRNA analysis to collect individual somata of dentate gyrus (DG) granule cells - the smallest neurons found in the brain - from living hippocampal slices of adult mice. We analyzed lipid extracts of each granule cell by nanoflow liquid chromatography (nano-LC) coupled to high-resolution time-of-flight mass spectrometry (MS). We were able reliably to detect many important lipids involved in membrane structure, energy storage, and cellular signaling. Importantly, we found that physiological stimulation of the lateral perforant path, a fiber tract that provides major excitatory input to DG granule cells, caused rapid and robust changes in the cells' lipid profile. The present application proposes to develop these initials findings into an optimized and validated protocol that can be widely applied to lipidomics analyses of neurons throughout the brain. We have two specific aims: (1) Method optimization. Our initial protocol is highly sensitive, but has three limits that stem from the vanishingly low amount of biomaterial afforded by a single neuron: (a) it covers only a fraction of the lipidome; (b) it allows tandem MS structure confirmation only for the most abundant lipid species; and (c) it provides relative rather than absolute quantification of detected lipids. We will (i) increase te sensitivity of our procedure through systematic modifications of key analytical parameters; (ii) extend the procedure's quantitative reach; and (iii) build reference libraries of lipid MS data for individual neurons, using pools of individually captured granule DG cells. (2) Method validation. Our preliminary work allowed us to identify a substantial number of lipid species in resting DG granule cells, and to detect specific alterations in the cells' lipid profile following physiologicl stimulation. To test the general applicability of the protocol, we will (i) profile the lipidome of pyramidal neurons in the CA1 and CA3 fields of the hippocampus, which are anatomically and functionally different from granule cells; and (ii) determine the impact of various physiological stimuli on the lipidome of DG granule cells and CA1/CA3 pyramidal neurons. Lastly, to define similarities and differences between single-cell and whole tissue preparations, we will compare the lipidomes of individual granule cells and micropunches of DG tissue under control and stimulated conditions. When fully validated and optimized, the present method will provide a flexible and robust new tool to investigate the roles of lipid molecules in identified neurons isolated from live brain tissue, opening exciting new avenues for research on neural lipids and the role of neuronal diversity in health and disease.


项目成果

期刊论文数量(3)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
A Primary Cortical Input to Hippocampus Expresses a Pathway-Specific and Endocannabinoid-Dependent Form of Long-Term Potentiation.
海马体的初级皮质输入表达了一种途径特异性和内源性大麻素依赖性的长期增强形式。
  • DOI:
    10.1523/eneuro.0160-16.2016
  • 发表时间:
    2016
  • 期刊:
  • 影响因子:
    3.4
  • 作者:
    Wang,Weisheng;Trieu,BrianH;Palmer,LindaC;Jia,Yousheng;Pham,DanielleT;Jung,Kwang-Mook;Karsten,CarleyA;Merrill,CollinB;Mackie,Ken;Gall,ChristineM;Piomelli,Daniele;Lynch,Gary
  • 通讯作者:
    Lynch,Gary
Author Correction: Patch clamp-assisted single neuron lipidomics.
作者更正:膜片钳辅助单神经元脂质组学。
  • DOI:
    10.1038/s41598-018-24605-7
  • 发表时间:
    2018
  • 期刊:
  • 影响因子:
    4.6
  • 作者:
    Merrill,CollinB;Basit,Abdul;Armirotti,Andrea;Jia,Yousheng;Gall,ChristineM;Lynch,Gary;Piomelli,Daniele
  • 通讯作者:
    Piomelli,Daniele
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Daniele Piomelli其他文献

Daniele Piomelli的其他文献

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{{ truncateString('Daniele Piomelli', 18)}}的其他基金

The lipid hydrolase NAAA as a target for non-addictive analgesic medications
脂质水解酶 NAAA 作为非成瘾性镇痛药物的靶标
  • 批准号:
    10584428
  • 财政年份:
    2023
  • 资助金额:
    $ 0.85万
  • 项目类别:
ICAL: Impact of Cannabinoids Across Lifespan
ICAL:大麻素对整个生命周期的影响
  • 批准号:
    10399921
  • 财政年份:
    2018
  • 资助金额:
    $ 0.85万
  • 项目类别:
ICAL: Impact of Cannabinoids Across Lifespan
ICAL:大麻素对整个生命周期的影响
  • 批准号:
    10188473
  • 财政年份:
    2018
  • 资助金额:
    $ 0.85万
  • 项目类别:
ICAL: Impact of Cannabinoids Across Lifespan: Administrative Core
ICAL:大麻素对整个生命周期的影响:管理核心
  • 批准号:
    10188474
  • 财政年份:
    2018
  • 资助金额:
    $ 0.85万
  • 项目类别:
ICAL: Impact of Cannabinoids Across Lifespan: Molecular Project
ICAL:大麻素对整个生命周期的影响:分子项目
  • 批准号:
    10188478
  • 财政年份:
    2018
  • 资助金额:
    $ 0.85万
  • 项目类别:
ICAL: Impact of Cannabinoids Across Lifespan
ICAL:大麻素对整个生命周期的影响
  • 批准号:
    10398657
  • 财政年份:
    2018
  • 资助金额:
    $ 0.85万
  • 项目类别:
Peripheral FAAH as a target for novel analgesics
外周 FAAH 作为新型镇痛药的靶点
  • 批准号:
    9454448
  • 财政年份:
    2017
  • 资助金额:
    $ 0.85万
  • 项目类别:
Peripheral FAAH as a target for novel analgesics
外周 FAAH 作为新型镇痛药的靶点
  • 批准号:
    9040444
  • 财政年份:
    2017
  • 资助金额:
    $ 0.85万
  • 项目类别:
A Protective role for 2-AG in age-dependent cognitive impairment.
2-AG 在年龄依赖性认知障碍中的保护作用。
  • 批准号:
    9180355
  • 财政年份:
    2016
  • 资助金额:
    $ 0.85万
  • 项目类别:
A Protective role for 2-AG in age-dependent cognitive impairment.
2-AG 在年龄依赖性认知障碍中的保护作用。
  • 批准号:
    9330759
  • 财政年份:
    2016
  • 资助金额:
    $ 0.85万
  • 项目类别:
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