Viscera receive innervation from sensory ganglia located adjacent to multiple levels of the brainstem and spinal cord. Here we examined whether molecular profiling could be used to identify functional clusters of colon afferents from thoracolumbar (TL), lumbosacral (LS), and nodose ganglia (NG) in male and female mice. Profiling of TL and LS bladder afferents was also performed. Visceral afferents were back-labeled using retrograde tracers injected into proximal and distal regions of colon or bladder, followed by single-cell qRT-PCR and analysis via an automated hierarchical clustering method. Genes were chosen for assay (32 for bladder; 48 for colon) based on their established role in stimulus detection, regulation of sensitivity/function, or neuroimmune interaction. A total of 132 colon afferents (from NG, TL, and LS ganglia) and 128 bladder afferents (from TL and LS ganglia) were analyzed. Retrograde labeling from the colon showed that NG and TL afferents innervate proximal and distal regions of the colon, whereas 98% of LS afferents only project to distal regions. There were clusters of colon and bladder afferents, defined by mRNA profiling, that localized to either TL or LS ganglia. Mixed TL/LS clustering also was found. In addition, transcriptionally, NG colon afferents were almost completely segregated from colon TL and LS neurons. Furthermore, colon and bladder afferents expressed genes at similar levels, although different gene combinations defined the clusters. These results indicate that genes implicated in both homeostatic regulation and conscious sensations are found at all anatomic levels, suggesting that afferents from different portions of the neuraxis have overlapping functions.

SIGNIFICANCE STATEMENT Visceral organs are innervated by sensory neurons whose cell bodies are located in multiple ganglia associated with the brainstem and spinal cord. For the colon, this overlapping innervation is proposed to facilitate visceral sensation and homeostasis, where sensation and pain are mediated by spinal afferents and fear and anxiety (the affective aspects of visceral pain) are the domain of nodose afferents. The transcriptomic analysis performed here reveals that genes implicated in both homeostatic regulation and pain are found in afferents across all ganglia types, suggesting that conscious sensation and homeostatic regulation are the result of convergence, and not segregation, of sensory input.

内脏从与脑干和脊髓的多个水平相邻的感觉神经节接受神经支配。在这里，我们检查了分子图谱是否可用于识别雄性和雌性小鼠胸腰椎（TL），腰ac部（LS）和结节神经节（NG）结肠传入性功能簇。还进行了TL和LS膀胱传入的分析。使用注入结肠或膀胱近端和远端区域的逆行示踪剂对内脏传入物质进行反标记，然后进行单细胞qRT-PCR并通过自动分层聚类方法进行分析。根据基因在刺激检测，敏感性/功能调节或神经免疫相互作用中已确立的作用，选择用于分析的基因（膀胱为32；结肠为48）。总共132个结肠传入（来自NG，TL，和LS神经节）和128个膀胱传入神经（来自TL和LS神经节）进行了分析。结肠的逆行标记显示NG和TL传入神经支配结肠的近端和远端区域，而98％的LS传入神经仅投射到远端区域。由mRNA谱图定义的结肠和膀胱传入簇位于TL或LS神经节。还发现了混合TL / LS群集。此外，在转录上，NG结肠传入几乎与结肠TL和LS神经元完全隔离。此外，尽管不同的基因组合定义了簇，但结肠和膀胱传入基因表达的基因水平相似。这些结果表明，在所有解剖学水平上都发现了与体内稳态调节和清醒感相关的基因，

重要声明内脏器官受感觉神经元支配，其细胞体位于与脑干和脊髓相关的多个神经节中。对于结肠，提出这种重叠的神经支配以促进内脏感觉和体内稳态，其中感觉和疼痛由脊髓传入介导，而恐惧和焦虑（内脏疼痛的情感方面）是结节传入的领域。此处进行的转录组分析显示，在所有类型的神经节的传入神经中都发现了与稳态调节和疼痛相关的基因，这表明有意识的感觉和稳态调节是感觉输入的趋同而非分离的结果。