Combinatorial Transcriptional Profiling of Mouse and Human Enteric Neurons Identifies Shared and Disparate Subtypes In Situ.
Combinatorial Transcriptional Profiling of Mouse and Human Enteric Neurons Identifies Shared and Disparate Subtypes In Situ.
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小鼠和人类肠神经元的组合转录谱分析在原位鉴定出共有的和不同的亚型。
DOI：
10.1053/j.gastro.2020.09.032
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发表时间：
2021-03
期刊：
影响因子：
29.4
通讯作者：
Southard-Smith EM
中科院分区：
文献类型：
--
作者：
May-Zhang AA;Tycksen E;Southard-Smith AN;Deal KK;Benthal JT;Buehler DP;Adam M;Simmons AJ;Monaghan JR;Matlock BK;Flaherty DK;Potter SS;Lau KS;Southard-Smith EM
肠神经系统（ENS）通过多种肠神经元（EN）的协同作用来协调肠道的基本功能。然而，目前缺乏关于肠神经元亚型完整的分子层面认知。为了比较人类和小鼠的肠神经元，我们对成年人类和小鼠健康的肠神经系统进行了转录组分析。我们旨在识别标记不同神经元亚型的转录本，并在多个肠道区域直观呈现人类和小鼠中保守的肠神经元亚型。
通过激光捕获显微切割技术分离人类肌间神经节及相邻的平滑肌，用于RNA测序。通过计算机减去肌肉基因特征来识别神经节特异性转录谱。分离小鼠肌间神经元的细胞核，并进行单核RNA测序（snRNA - Seq），共获得超过40亿条读数和25208个神经元。利用小鼠单核RNA测序数据定义神经元亚型。对人类和小鼠数据集进行比较信息学分析，确定共享的肠神经元亚型标记物，并使用杂交链式反应（HCR）进行原位可视化。
基于同源基因表达情况，人类和小鼠十二指肠、回肠和结肠中的几种肠神经元亚型是保守的。然而，小鼠中一些肠神经元亚型特异性基因在人类中表达于形态学上完全不同的亚型。在小鼠中，我们鉴定出几种神经元亚型，它们在所有肠段中稳定表达基因模块，且一种或多种标记基因呈现梯度性、区域性表达。
我们对人类肌间神经节和小鼠肠神经元的联合转录组分析，为开发新型肠道疗法奠定了丰富的基础。部分肠神经元亚型存在一致性，但我们也注意到多种物种差异，因此在将小鼠肠神经系统研究结果应用于人类胃肠道研究时，应谨慎考虑这些差异。
The enteric nervous system (ENS) coordinates essential intestinal functions through the concerted action of diverse enteric neurons (EN). However, integrated molecular knowledge of EN subtypes is lacking. To compare human and mouse ENs, we transcriptionally profiled healthy ENS from adult humans and mice. We aimed to identify transcripts marking discrete neuron subtypes and visualize conserved EN subtypes for humans and mice in multiple bowel regions. Human myenteric ganglia and adjacent smooth muscle were isolated by laser-capture microdissection for RNA-Seq. Ganglia-specific transcriptional profiles were identified by computationally subtracting muscle gene signatures. Nuclei from mouse myenteric neurons were isolated and subjected to single-nucleus RNA-Seq (snRNA-Seq), totaling over four billion reads and 25,208 neurons. Neuronal subtypes were defined using mouse snRNA-Seq data. Comparative informatics between human and mouse datasets identified shared EN subtype markers, which were visualized in situ using hybridization chain reaction (HCR). Several EN subtypes in the duodenum, ileum, and colon are conserved between humans and mice based on orthologous gene expression. However, some EN subtype-specific genes from mice are expressed in completely distinct morphologically defined subtypes in humans. In mice, we identified several neuronal subtypes that stably express gene modules across all intestinal segments, with graded, regional expression of one or more marker genes. Our combined transcriptional profiling of human myenteric ganglia and mouse EN provides a rich foundation for developing novel intestinal therapeutics. There is congruency among some EN subtypes, but we note multiple species differences that should be carefully considered when relating findings from mouse ENS research to human GI studies.
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影响因子：
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