Deep sequencing of Phox2a nuclei reveals five classes of anterolateral system neurons-猫眼课题宝

DOI：

10.1101/2023.08.20.553715

发表时间：

2023-09

期刊：

bioRxiv

影响因子：

通讯作者：

中科院分区：

其他

文献类型：

作者：

Andrew M. Bell;Charlotte Utting;A. Dickie;M. Kucharczyk;Raphaëlle Quillet;M. Gutierrez-Mecinas;Aimi N B Razlan;A. Cooper;Yuxuan Lan;J. Hachisuka;Greg A Weir;K. Bannister;Masahiko Watanabe;Artur Kania;M. Hoon;I. Macaulay;Franziska Denk;A. Todd

前外侧系统（ALS）是从脊髓投射到多个大脑区域的主要上行通路，并且是疼痛、瘙痒和皮肤温度感知的基础。尽管它的重要性，我们对这个系统的理解一直受到其组成细胞的功能和分子多样性的阻碍。在这里，我们使用荧光激活细胞分选分离ALS神经元属于Phox 2a-谱系的单核RNA测序。我们揭示了五个不同的集群ALS神经元（ALS 1 -5）和文件的层状分布在脊髓中使用原位杂交。我们鉴定出3个主要位于背角I-III层的神经元簇（ALS 1 -3）和两个位于更深层的细胞体簇（ALS 4和ALS 5）。我们的研究结果揭示了成年小鼠ALS神经元多样性的转录逻辑，并揭示了两个先前确定的投射神经元类别的分子身份。我们还表明，这些分子特征可以用于使用逆行病毒追踪靶向ALS神经元组。总的来说，我们的研究结果为研究体感生物学和针对ALS神经元亚类提供了宝贵的资源。前外侧系统（ALS）是脊髓的一个主要上行通路，是感知疼痛、瘙痒和皮肤温度的基础。因此，它是开发慢性疼痛新疗法的重要目标。我们对这个系统的理解一直受到其组成细胞的相当大的多样性的阻碍。在这里，我们通过使用高分辨率RNA测序来剖析这些细胞的复杂异质性。我们揭示了五种不同类型的ALS神经元，它们在脊髓内的差异分布，并可能代表功能群体。我们的数据提供了新的见解ALS的分子结构，并将是重要的未来研究，以确定不同类型的ALS细胞在感觉处理中的作用。

The anterolateral system (ALS) is a major ascending pathway from the spinal cord that projects to multiple brain areas and underlies the perception of pain, itch and skin temperature. Despite its importance, our understanding of this system has been hampered by the considerable functional and molecular diversity of its constituent cells. Here we use fluorescence-activated cell sorting to isolate ALS neurons belonging to the Phox2a-lineage for single-nucleus RNA sequencing. We reveal five distinct clusters of ALS neurons (ALS1-5) and document their laminar distribution in the spinal cord using in situ hybridization. We identify 3 clusters of neurons located predominantly in laminae I-III of the dorsal horn (ALS1-3) and two clusters with cell bodies located in deeper laminae (ALS4 & ALS5). Our findings reveal the transcriptional logic that underlies ALS neuronal diversity in the adult mouse and uncover the molecular identity of two previously identified classes of projection neurons. We also show that these molecular signatures can be used to target groups of ALS neurons using retrograde viral tracing. Overall, our findings provide a valuable resource for studying somatosensory biology and targeting subclasses of ALS neurons. Significance Statement The anterolateral system (ALS) is a major ascending pathway from the spinal cord that underlies perception of pain, itch and skin temperature. It is therefore an important target for the development of new treatments for chronic pain. Our understanding of this system has been hampered by the considerable diversity of its constituent cells. Here we dissect the complex heterogeneity of these cells by using high-resolution RNA sequencing. We reveal five distinct types of ALS neurons, which are differentially distributed within the spinal cord, and probably represent functional populations. Our data provide novel insights into the molecular architecture of the ALS, and will be important for future studies to define the roles of different ALS cell types in sensory processing.

Deep sequencing of Phox2a nuclei reveals five classes of anterolateral system neurons

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