Tools for analysis and conditional deletion of subsets of sensory neurons
Tools for analysis and conditional deletion of subsets of sensory neurons
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.
Santana-Varela, Sonia
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Bogdanov, Yury
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Gossage, Samuel J.
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Okorokov, Andrei L.
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Li, Shengnan
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de Clauser, Larissa
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Alves-Simoes, Marta
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Sexton, Jane E.
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Iseppon, Federico
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Luiz, Ana P.
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30 September 2021
Santana-Varela, Sonia
4694b021-abaf-4b1b-a561-23e63f076c3b
Bogdanov, Yury
0c970999-e191-4f1b-90d9-7bf25a5d5b4b
Gossage, Samuel J.
8c32a048-2e69-44f3-a1e5-3844f9461520
Okorokov, Andrei L.
0e171d26-1418-4955-b8ce-f89543129ec1
Li, Shengnan
0f2f3693-9107-4055-bfbc-f8016323ea01
de Clauser, Larissa
8be9ea0d-9bd9-4e2b-8cac-0476f2c4817b
Alves-Simoes, Marta
802425d0-4783-4755-8f8a-df6895243820
Sexton, Jane E.
92a69ad9-8ac3-45a8-80e4-148f2e727359
Iseppon, Federico
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Luiz, Ana P.
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Santana-Varela, Sonia, Bogdanov, Yury, Gossage, Samuel J., Okorokov, Andrei L., Li, Shengnan, de Clauser, Larissa, Alves-Simoes, Marta, Sexton, Jane E., Iseppon, Federico and Luiz, Ana P.
(2021)
Tools for analysis and conditional deletion of subsets of sensory neurons.
Wellcome Open Research, 6.
(doi:10.12688/wellcomeopenres.17090.1).
Abstract
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.
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Accepted/In Press date: 6 September 2021
Published date: 30 September 2021
Identifiers
Local EPrints ID: 452743
URI: http://eprints.soton.ac.uk/id/eprint/452743
ISSN: 2398-502X
PURE UUID: 9ed48b2f-7489-4313-841a-7044e21e29b4
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Date deposited: 17 Dec 2021 17:51
Last modified: 17 Mar 2024 03:37
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Contributors
Author:
Sonia Santana-Varela
Author:
Samuel J. Gossage
Author:
Andrei L. Okorokov
Author:
Shengnan Li
Author:
Larissa de Clauser
Author:
Marta Alves-Simoes
Author:
Jane E. Sexton
Author:
Federico Iseppon
Author:
Ana P. Luiz
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