A novel method to sort and enrich sensory neurons.

IF 2.3 4区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Zerina Kurtović, Sven David Arvidsson, Juan Antonio Vazquez Mora, Sijing Ye, Alex Bersellini Farinotti, Nils Simon, Emerson Krock, Lisbet Haglund, Michael Hagemann-Jensen, Harald Lund, Camilla I Svensson
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Abstract

Peripheral sensory neurons, residing in the dorsal root ganglia (DRG), relay sensory information from the periphery to the central nervous system. Although single-cell transcriptomic studies have identified over 20 distinct sensory neuron subtypes, functional analysis and assessment of subtype-specific pathological changes remain difficult. Effective isolation and enrichment of sensory neurons are challenging yet essential for functional studies. Therefore, we used single-cell transcriptomic data from DRG to identify a panel of neuronal surface markers, including Nrxn2 and Pirt. Using these markers, we developed a fluorescence-activated cell sorting (FACS) panel for neuronal enrichment and analysis that does not rely on transgenic mouse strains and can be broadly applied. The panel was validated by microscopy and single-cell RNA (scRNA) sequencing, which also revealed broad representation of neuronal subtypes. Expression of these markers in human DRG underscores the translational value of this isolation method for sensory and pain studies. Overall, this study provides a valuable tool for isolating DRG neurons, advancing research on sensory neuron function and pain biology, and facilitating neuroimmune studies.

一种分类和丰富感觉神经元的新方法。
位于背根神经节(DRG)的外周感觉神经元将外周神经系统的感觉信息传递给中枢神经系统。尽管单细胞转录组学研究已经确定了20多种不同的感觉神经元亚型,但对亚型特异性病理变化的功能分析和评估仍然很困难。有效地分离和富集感觉神经元对功能研究具有挑战性,但也是必不可少的。因此,我们使用来自DRG的单细胞转录组数据来鉴定一组神经元表面标记,包括Nrxn2和Pirt。利用这些标记,我们开发了一种荧光激活细胞分选(FACS)面板,用于神经元富集和分析,不依赖于转基因小鼠品系,可以广泛应用。该小组通过显微镜和单细胞RNA (scRNA)测序进行了验证,这也揭示了神经元亚型的广泛代表性。这些标记物在人类DRG中的表达强调了这种分离方法在感觉和疼痛研究中的转化价值。总之,本研究为分离DRG神经元、推进感觉神经元功能和疼痛生物学研究以及促进神经免疫研究提供了有价值的工具。
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来源期刊
Journal of Neuroscience Methods
Journal of Neuroscience Methods 医学-神经科学
CiteScore
7.10
自引率
3.30%
发文量
226
审稿时长
52 days
期刊介绍: The Journal of Neuroscience Methods publishes papers that describe new methods that are specifically for neuroscience research conducted in invertebrates, vertebrates or in man. Major methodological improvements or important refinements of established neuroscience methods are also considered for publication. The Journal''s Scope includes all aspects of contemporary neuroscience research, including anatomical, behavioural, biochemical, cellular, computational, molecular, invasive and non-invasive imaging, optogenetic, and physiological research investigations.
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