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Urocortin3-expressing neurons in sensory transmission
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology. (Malin Lagerström)ORCID iD: 0000-0002-2174-6392
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Genomics and Neurobiology. (Malin Lagerström)ORCID iD: 0000-0003-4095-6456
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Genomics and Neurobiology. (Malin Lagerström)ORCID iD: 0000-0002-9681-5129
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Genomics and Neurobiology. (Malin Lagerström)
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Urocortin 3 (UCN3) is a neuropeptide involved in mechanosensation and stress regulation, and Ucn3-Cre neurons have been assigned a role in mechanical itch. Here, we show that Ucn3 marks a population of excitatory neurons in the mouse dorsal horn, divided into two non-overlapping subpopulations expressing protein kinase C g or calretinin/calbindin 2. Chemogenetic activation of spinal Ucn3-Cre neurons evoked a targeted biting/licking behavior towards the corresponding dermatome. Genetic deletion of vesicular glutamate transporter 2 (VGLUT2) in Ucn3-Cre neurons removed the phenotype, showing that the biting/licking behavior is VGLUT2-dependent. Conditional deletion of VGLUT2 did not affect acute thermal or mechanical withdrawal responses, nor thermal withdrawal responses after nerve growth factor-induced hypersensitivity or the prurifensive response to 48/80 or von Frey stimuli applied in nape. Instead, we found that a group of spinal Ucn3 neurons were activated in response to artificial scratching or 48/80-induced itch. Electrophysiological experiments showed that spinal Ucn3 neurons received both glycinergic and GABAergic tonic inhibition, and monosynaptic inputs from both Aβ and C fibers, which could be confirmed by rabies tracing. Spinal Ucn3/Ucn3-Cre neurons thus represent a mechanically sensitive population with several roles in the itch-scratch cycle. 

Keywords [en]
Urocortin 3, spinal cord, itch, vesicular glutamate transporter 2, retrograde rabies tracing, pharmacogenetics
National Category
Neurosciences
Identifiers
URN: urn:nbn:se:uu:diva-499112OAI: oai:DiVA.org:uu-499112DiVA, id: diva2:1745462
Funder
Swedish Research Council, 2015-02442Swedish Research Council, 2016-00851The Swedish Brain FoundationKnut and Alice Wallenberg Foundation, 2019.0047Swedish Society for Medical Research (SSMF), P14-0033Olle Engkvists stiftelse, 190-0370Gunvor och Josef Anérs stiftelseO.E. och Edla Johanssons vetenskapliga stiftelseThe Royal Swedish Academy of SciencesUppsala UniversityAvailable from: 2023-03-23 Created: 2023-03-23 Last updated: 2023-03-23
In thesis
1. Somatosensory Circuits in the Central Nervous System
Open this publication in new window or tab >>Somatosensory Circuits in the Central Nervous System
2023 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Spinal glycinergic neurons prevent spontaneous sensations of pain and itch, and development of persistent pain and itch conditions. The glycine receptor alpha 3 subunit (GLRA3) is suggested to be involved in certain inflammatory models, but not in acute nociception. In paper I, we found that spinal Glra3-Cre neurons mediate compound 48/80- and chloroquine-evoked itch. This population receive monosynaptic input from local spinal neurons, motor- and sensory-associated brain areas and innocuous, proprioceptive, noxious and pruritic primary afferents. In paper II, global exonal deletion of the Vglut2 or Viaat locus in Glra3-Cre neurons did not affect the withdrawal response to interleukin 1 beta-induced thermal hypersensitivity. In paper III, single-cell RNA sequencing and bulk qRT-PCR analyses revealed widespread, but low Glra3 expression in both Slc17a6(+) and Slc32a1(+) neurons within the central nervous system. Males expressed higher levels in all investigated brain structures compared to females and RNAscope spatially validated Glra3 expression in Slc17a6(+) and Slc32a1(+) neurons. Urocortin 3 (UCN3) is a neuropeptide belonging to the corticotropin-releasing factor family. The spinal KF43 Ucn3-Cre neurons mediate mechanical itch and alloknesis. In paper IV, the lumbar KF31 Ucn3-Cre neurons were divided into two non-overlapping protein kinase C gamma(+) and calretinin(+) subpopulations. Activation evoked biting/licking and Cre-dependent exonal deletion of the Vglut2 locus removed this phenotype. The deletion did not affect the response to mechanical stimulation nor compound 48/80, however, these sensory-modality activated fos(+) cells co-expressed Ucn3. In rodents, the layer IV barrel field of S1 comprises of excitatory pyramidal and spiny stellate cells and this area receives sensory input from the whisker pad via the thalamic ventroposteromedial nucleus. Vesicular monoaminergic transporter 2 (VMAT2) sequesters monoamines into synaptic vesicles. Vmat2 is prominent in the developmental brain, while more sparsely detected in the adult brain. In paper V, a Vmat2-Cre mouse line labelled adult Vmat2(+) and Cre(+) layer IV S1 barrel neurons, which received monosynaptic thalamic input. These neurons displayed a homogenous intrinsically bursting firing pattern, indicative of excitatory neurons and shared the morphology of stellate spiny neurons. The Vmat2-Cre neurons also expressed excitatory and layer IV markers, and mainly cluster with layer IV neurons.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2023. p. 95
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1929
Keywords
Glycine receptor alpha 3, urocortin 3, vesicular monoaminergic transporter 2, nociception, pruriception, spinal cord, somatosensory cortex, DREADD, rabies tracing, transcriptomics
National Category
Neurosciences
Research subject
Neuroscience
Identifiers
urn:nbn:se:uu:diva-499118 (URN)978-91-513-1768-7 (ISBN)
Public defence
2023-05-17, Room A1:111, BMC, Husargatan 3, Uppsala, 13:15 (English)
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Supervisors
Available from: 2023-04-25 Created: 2023-03-23 Last updated: 2023-04-25

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Franck, Marina Christina MikaelaMa, HaishaKoning, Harmen KornelisÖhman, Caroline

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Franck, Marina Christina MikaelaWeman, Hannah M.Ceder, Mikaela M.Ahemaiti, AikeremuMa, HaishaMagnusson, Kajsa A.Koning, Harmen KornelisÖhman, CarolineLagerström, Malin C.
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