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Deep FLASH-seq profiling of purified canine sensory neurons uncovers species-specific signatures relevant to pain and itch

ABSTRACT: Naturally occurring pain and itch disorders in the domestic dog represent an important and underexploited opportunity for translational sensory neuroscience. These conditions affect companion animals in a manner that largely mirrors human disease, highlighting the need for a detailed comparative understanding of canine somatosensory neurobiology. Here, we present a single-cell transcriptomic characterisation of the canine dorsal root ganglion (DRG), generated to provide molecular insights into sensory neuron diversity in a species of direct veterinary and biomedical relevance. We develop a novel mechanical dissociation and fluorescence-activated cell sorting strategy that enables purification of intact whole neurons from adult canine DRG, followed by deep, full-length RNA sequencing using FLASH-seq. This approach yields high-quality transcriptional profiles with molecular depth analogous to deep neuronal profiling approaches applied in human DRG, enabling resolution of detailed neuronal identities and subtype-specific gene programs. Using these data, we identify a diverse repertoire of canine sensory neuron clusters that conform to conserved principles of DRG molecular organization observed across species, including peptidergic and non-peptidergic nociceptors, low-threshold mechanoreceptors, proprioceptors, and thermosensory populations. Leveraging the depth of the transcriptomic data, we perform cross-species comparisons with human and mouse DRG datasets, revealing broad conservation of pain- and itch-relevant pathways and therapeutic targets, alongside biologically meaningful divergence. We further identify species-specific differences in neuronal subtype-restricted expression of the pharmacologically relevant receptors IL31RA and SSTR2, which we validate using in situ hybridization in canine DRG tissue and contextualize using human spatial transcriptomic data. Finally, we provide evidence that genes implicated in canine domestication are non-randomly enriched in specific sensory neuron populations, suggesting that evolutionary history may have shaped aspects of somatosensory function. Overall, these data represent a resource for comparative sensory neuroscience and inform translational interpretation of pain and itch therapeutics across species.

ORGANISM(S): Canis lupus familiaris

PROVIDER: GSE327952 | GEO | 2026/08/18

REPOSITORIES: GEO

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Project description:Sensory neurons are distinguished by distinct signaling networks and receptive characteristics. Thus, sensory neuron types can be defined by linking transcriptome-based neuron typing with the sensory phenotypes. Here we classify somatosensory neurons of the mouse dorsal root ganglion (DRG) by high-coverage single-cell RNA-sequencing (10 950 Â± 1 218 genes per neuron) and neuron size-based hierarchical clustering. Moreover, single DRG neurons responding to cutaneous stimuli are recorded using an in vivo whole-cell patch clamp technique and classified by neuron-type genetic markers. Small diameter DRG neurons are classified into one type of low-threshold mechanoreceptor and five types of mechanoheat nociceptors (MHNs). Each of the MHN types is further categorized into two subtypes. Large DRG neurons are categorized into four types, including neurexophilin 1-expressing MHNs and mechanical nociceptors (MNs) expressing BAI1-associated protein 2-like 1 (Baiap2l1). Mechanoreceptors expressing trafficking protein particle complex 3-like and Baiap2l1-marked MNs are subdivided into two subtypes each. These results provide a new system for cataloging somatosensory neurons and their transcriptome databases. RNA-seq of mRNA levels in 197 individual DRG neurons We performed RNA-seq on total 203 individual DRG neurons. Six of them were not qualified and thus, were excluded for further analysis. To evaluate the quality of RNA-seq, we randomly devided No.72 neurons into two parts and performed RNA-seq seperately. Thus, we had 204 individual samples from 203 individual DRG and 198 individual qualified samples from 197 individual DRG. To evaluate the homogeneity of RNA-seq data from different mice at the same age just as used, we performed RNA-seq on 5 single DRG from different mice. Here, these data from DRG were also considered as experimental control. The 'DRG_neurons_RNA_Seq.txt' contains processed data for 204 samples and 'DRG_RNA_Seq.txt' for 5 samples.

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Deep FLASH-seq profiling of purified canine sensory neurons uncovers species-specific signatures relevant to pain and itch

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