Tools for analysis and conditional deletion of subsets of sensory neurons.
Tools for analysis and conditional deletion of subsets of sensory neurons.
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DOI：
10.12688/wellcomeopenres.17090.1
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发表时间：
2021
影响因子：
--
通讯作者：
Cox JJ
中科院分区：
文献类型：
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作者：
Santana-Varela S;Bogdanov YD;Gossage SJ;Okorokov AL;Li S;de Clauser L;Alves-Simoes M;Sexton JE;Iseppon F;Luiz AP;Zhao J;Wood JN;Cox JJ
背景：躯体感觉依赖于三叉神经和背根神经节（DRG）的初级感觉神经元。小鼠DRG感觉神经元的转录谱已经定义了至少18种不同的神经元细胞类型。使用advillin启动子，我们已经产生了一个转基因小鼠系，只表达白喉毒素A（DTA）在感觉神经元中的Cre重组酶的存在下。这使得我们能够使用一系列已建立的和新的Cre系（包括所有感觉神经元）消融DRG内的特定神经元亚群。 研究方法：在小鼠advillin DRG启动子的控制下，产生了一个双链-tdTomato-stop-DTA细菌人工染色体（BAC）转基因报告细胞系（AdvDTA）。首先使用Na v1.8 Cre验证该细胞系，然后与CGRP CreER（Calca）、Th CreERT 2、Tmem 45 b Cre、Tmem 233 Cre、Ntng 1 Cre和TrkB CreER（Ntrk 2）细胞系杂交。疼痛行为测定包括Hargresten、热板、Randall-Selitto、冷足底、部分坐骨神经结扎和福尔马林试验。 结果：运动活动，通过旋转棒测试评估，是正常的所有线测试。当Na v1.8阳性神经元或Tmem 45 b阳性神经元被消融时，伤害性机械感觉显著降低，而急性热痛不受影响。与此相反，伤害性机械感觉是正常的CGRP阳性神经元消融后，但急性热痛阈值显着升高，减少疼痛反应中观察到的福尔马林试验的第二阶段。在部分坐骨神经结扎神经病理性疼痛模型中，TrkB阳性神经元的消融导致机械超敏反应的显著缺陷。 结论：使用AdvDTA线消融特定DRG神经元亚群将是进一步表征躯体感觉处理、神经免疫相互作用和慢性疼痛障碍的有用资源。
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 Na v1.8 Cre and then crossed to CGRP CreER (Calca), Th CreERT2, Tmem45b Cre, Tmem233 Cre, Ntng1 Cre and TrkB CreER (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 Na v1.8 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.