Profiling 26,000 Aplysia californica neurons by single cell mass spectrometry reveals neuronal populations with distinct neuropeptide profiles.
Profiling 26,000 Aplysia californica neurons by single cell mass spectrometry reveals neuronal populations with distinct neuropeptide profiles.
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DOI：
10.1016/j.jbc.2022.102254
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发表时间：
2022-08
影响因子：
4.8
通讯作者：
Sweedler, Jonathan, V
中科院分区：
文献类型：
--
作者：
Chan-Andersen, Peter C.;Romanova, Elena, V;Rubakhin, Stanislav S.;Sweedler, Jonathan, V
神经肽是一类化学性质多样的细胞间信号传导分子，通常以细胞特异性方式在整个中枢神经系统中广泛表达。虽然神经肽的细胞与细胞之间的差异是预期的，但通常不清楚神经肽的表达在神经元之间究竟如何变化。在这里，我们创建了一个显微镜引导的，高通量的单细胞基质辅助激光解吸/电离质谱法的方法来调查的神经生物学模型的神经肽的异质性的个体神经元的中枢神经系统的加利福尼亚州，加州海兔。总之，我们分析了来自18只动物的26，000多个神经元，并通过与从UniProt数据库检索的已知Aaplasia前激素生成的计算机肽库进行质量匹配，分配了来自66种前激素的866种肽。Louvain-Jaccard（LJ）对来自单个神经元的质谱进行聚类，揭示了40个独特的神经元群体或LJ簇，每个具有不同的神经肽谱。激素原及其相关肽通常存在于神经节的单个细胞中，这与激素原先前已知的神经节定位一致。几个LJ簇也揭示了行为相关的激素原的细胞共定位，如LJ簇表现出参与摄食的无毛蛋白和神经肽Y，和另一个簇，其特征在于尾加压素II，小心脏肽，传感蛋白A，和FRFa，其在摄食网络中显示出活性或存在于摄食肌肉组织中。这种基于质谱的方法能够基于单细胞神经肽含量对大细胞群进行稳健的分类，并且易于适用于一系列动物和组织类型的研究。
Neuropeptides are a chemically diverse class of cell-to-cell signaling molecules that are widely expressed throughout the central nervous system, often in a cell-specific manner. While cell-to-cell differences in neuropeptides is expected, it is often unclear how exactly neuropeptide expression varies among neurons. Here we created a microscopy-guided, high-throughput single cell matrix-assisted laser desorption/ionization mass spectrometry approach to investigate the neuropeptide heterogeneity of individual neurons in the central nervous system of the neurobiological model Aplysia californica, the California sea hare. In all, we analyzed more than 26,000 neurons from 18 animals and assigned 866 peptides from 66 prohormones by mass matching against an in silico peptide library generated from known Aplysia prohormones retrieved from the UniProt database. Louvain–Jaccard (LJ) clustering of mass spectra from individual neurons revealed 40 unique neuronal populations, or LJ clusters, each with a distinct neuropeptide profile. Prohormones and their related peptides were generally found in single cells from ganglia consistent with the prohormones’ previously known ganglion localizations. Several LJ clusters also revealed the cellular colocalization of behaviorally related prohormones, such as an LJ cluster exhibiting achatin and neuropeptide Y, which are involved in feeding, and another cluster characterized by urotensin II, small cardiac peptide, sensorin A, and FRFa, which have shown activity in the feeding network or are present in the feeding musculature. This mass spectrometry–based approach enables the robust categorization of large cell populations based on single cell neuropeptide content and is readily adaptable to the study of a range of animals and tissue types.
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影响因子：
7.4
作者：
Do TD;Comi TJ;Dunham SJ;Rubakhin SS;Sweedler JV
通讯作者：
Sweedler JV
DOI：
10.1007/978-1-4939-7537-2_17
发表时间：
2018-01-01
期刊：
PEPTIDOMICS: METHODS AND STRATEGIES
影响因子：
--
作者：
DeLaney, Kellen;Buchberger, Amanda;Li, Lingjun
通讯作者：
Li, Lingjun
DOI：
10.1002/cphc.201701364
发表时间：
2018-05-22
期刊：
Chemphyschem : a European journal of chemical physics and physical chemistry
影响因子：
--
作者：
Do TD;Ellis JF;Neumann EK;Comi TJ;Tillmaand EG;Lenhart AE;Rubakhin SS;Sweedler JV
通讯作者：
Sweedler JV
影响因子：
3.4
作者：
Birner-Gruenberger, Ruth;Darnhofer, Barbara;Lubec, Gert
通讯作者：
Lubec, Gert
DOI：
10.1021/jasms.0c00191
发表时间：
2020-12-02
影响因子：
3.2
作者：
DeLaney K;Li L
通讯作者：
Li L