Tools for analysis and conditional deletion of subsets of sensory neurons

Wellcome Open Research

article

Sonia Santana-Varela¹  Yury D. Bogdanov¹  Samuel J. Gossage¹  Andrei L. Okorokov¹  Shengnan Li¹  Larissa de Clauser¹  Marta Alves-Simoes¹  Jane E. Sexton¹  Federico Iseppon¹  Ana P. Luiz¹  Jing Zhao¹  John N. Wood¹  James J. Cox¹ 

[1] Molecular Nociception Group, University College London;Antibody and Vaccine Group, Centre for Cancer Immunology, MP127, University of Southampton Faculty of Medicine;Institute for Biomedicine, Affiliated Institute of the University of Lubeck

关键词: Advillin;    dorsal root ganglia;    primary sensory neuron;    pain;    somatosensation;    Cre recombinase;    diphtheria toxin;   

DOI  :  10.12688/wellcomeopenres.17090.1

学科分类：内科医学

来源: Wellcome

【 摘 要 】

Background: Somatosensation depends on primary sensory neurons of the trigeminal and dorsal root ganglia (DRG). Transcriptional profiling of mouse DRG sensory neurons has defined at least 18 distinct neuronal cell types. Using an advillin promoter, we have generated a transgenic mouse line that only expresses diphtheria toxin A (DTA) in sensory neurons in the presence of Cre recombinase. This has allowed us to ablate specific neuronal subsets within the DRG using a range of established and novel Cre lines that encompass all sets of sensory neurons.   Methods: A floxed-tdTomato-stop-DTA bacterial artificial chromosome (BAC) transgenic reporter line (AdvDTA) under the control of the mouse advillin DRG promoter was generated. The line was first validated using a Nav1.8Cre and then crossed to CGRPCreER (Calca), ThCreERT2, Tmem45bCre, Tmem233Cre, Ntng1Cre and TrkBCreER (Ntrk2) lines. Pain behavioural assays included Hargreaves’, hot plate, Randall-Selitto, cold plantar, partial sciatic nerve ligation and formalin tests.Results: Motor activity, as assessed by the rotarod test, was normal for all lines tested. Noxious mechanosensation was significantly reduced when either Nav1.8 positive neurons or Tmem45b positive neurons were ablated whilst acute heat pain was unaffected. In contrast, noxious mechanosensation was normal following ablation of CGRP-positive neurons but acute heat pain thresholds were significantly elevated and a reduction in nocifensive responses was observed in the second phase of the formalin test. Ablation of TrkB-positive neurons led to significant deficits in mechanical hypersensitivity in the partial sciatic nerve ligation neuropathic pain model.Conclusions: Ablation of specific DRG neuronal subsets using the AdvDTA line will be a useful resource for further functional characterization of somatosensory processing, neuro-immune interactions and chronic pain disorders.

【 授权许可】

CC BY