Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment.
Single nuclei RNA sequencing of the rat AP and NTS following GDF15 treatment.
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DOI：
10.1016/j.molmet.2021.101422
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发表时间：
2022-03
影响因子：
8.1
通讯作者：
De Jonghe BC
中科院分区：
文献类型：
--
作者：
Reiner BC;Crist RC;Borner T;Doyle RP;Hayes MR;De Jonghe BC
已知生长分化因子15（GDF 15）在进食、恶心和体重中发挥作用，其作用通过最后区（AP）和孤束核（NTS）中的GFRAL-RET受体复合物进行。为了进一步阐明GDF 15信号传导下游的潜在细胞类型特异性分子机制，我们使用单核RNA测序（snRNAseq）方法来分析全身性GDF 15治疗后AP和NTS细胞亚型特异性转录组。在GDF 15或盐水注射后6小时，AP和NTS微穿孔用于来自Sprague道利大鼠的snRNAseq，并且Seurat用于鉴定由于全身性GDF 15处理的直接和继发效应引起的基因表达中的细胞亚型和细胞类型特异性改变。使用135，000个单独AP/NTS核的转录组谱，我们鉴定了19个转录组学上不同的细胞亚型，包括单个群体Gfral和Ret阳性兴奋性神经元，其代表GDF 15的主要作用位点。在神经元和神经胶质中鉴定了总共10600个细胞类型特异性差异表达事件，包括鉴定Gfral-Ret阳性兴奋性神经元中GDF 15直接作用特异性的转录组改变以及神经元和神经胶质细胞类型之间共有的转录组改变。下游分析确定了所观察到的转录组改变的信号通路和上游调控机制中的共享和细胞类型特异性改变。这些数据为我们理解AP和NTS细胞类型特异性的与GDF 15信号相关的分子机制提供了相当大的进展。所确定的细胞亚型特异性调节机制和信号通路可能代表未来药物治疗的重要靶点。GDF 15直接改变Gfral和Ret阳性兴奋性神经元中的转录。GDF 15间接改变其他神经元和神经胶质细胞群体中的转录。细胞类型特异性表达变化识别调控和信号传导机制。
Growth differentiation factor 15 (GDF15) is known to play a role in feeding, nausea, and body weight, with action through the GFRAL-RET receptor complex in the area postrema (AP) and nucleus tractus solitarius (NTS). To further elucidate the underlying cell type-specific molecular mechanisms downstream of GDF15 signaling, we used a single nuclei RNA sequencing (snRNAseq) approach to profile AP and NTS cellular subtype-specific transcriptomes after systemic GDF15 treatment. AP and NTS micropunches were used for snRNAseq from Sprague Dawley rats 6 h following GDF15 or saline injection, and Seurat was used to identify cellular subtypes and cell type-specific alterations in gene expression that were due to the direct and secondary effects of systemic GDF15 treatment. Using the transcriptome profile of ∼35,000 individual AP/NTS nuclei, we identified 19 transcriptomically distinct cellular subtypes, including a single population Gfral and Ret positive excitatory neurons, representing the primary site of action for GDF15. A total of ∼600 cell type-specific differential expression events were identified in neurons and glia, including the identification of transcriptome alterations specific to the direct effects of GDF15 in the Gfral-Ret positive excitatory neurons and shared transcriptome alterations across neuronal and glial cell types. Downstream analyses identified shared and cell type-specific alterations in signaling pathways and upstream regulatory mechanisms of the observed transcriptome alterations. These data provide a considerable advance in our understanding of AP and NTS cell type-specific molecular mechanisms associated with GDF15 signaling. The identified cellular subtype-specific regulatory mechanism and signaling pathways likely represent important targets for future pharmacotherapies. GDF15 directly alters transcription in Gfral- and Ret-positive excitatory neurons. GDF15 indirectly alters transcription in other neuronal and glial populations. Cell type-specific expression changes identify regulatory and signaling mechanisms.
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