PMID- 32079727
OWN - NLM
STAT- MEDLINE
DCOM- 20200713
LR  - 20240328
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 117
IP  - 10
DP  - 2020 Mar 10
TI  - An integrative approach to the facile functional classification of dorsal root 
      ganglion neuronal subclasses.
PG  - 5494-5501
LID - 10.1073/pnas.1911382117 [doi]
AB  - Somatosensory neurons have historically been classified by a variety of 
      approaches, including structural, anatomical, and genetic markers; 
      electrophysiological properties; pharmacological sensitivities; and more 
      recently, transcriptional profile differentiation. These methodologies, used 
      separately, have yielded inconsistent classification schemes. Here, we describe 
      phenotypic differences in response to pharmacological agents as measured by 
      changes in cytosolic calcium concentration for the rapid classification of 
      neurons in vitro; further analysis with genetic markers, whole-cell recordings, 
      and single-cell transcriptomics validated these findings in a functional context. 
      Using this general approach, which we refer to as tripartite constellation 
      analysis (TCA), we focused on large-diameter dorsal-root ganglion (L-DRG) neurons 
      with myelinated axons. Divergent responses to the K-channel antagonist, 
      kappaM-conopeptide RIIIJ (RIIIJ), reliably identified six discrete functional cell 
      classes. In two neuronal subclasses (L1 and L2), block with RIIIJ led to an 
      increase in [Ca] (i) Simultaneous electrophysiology and calcium imaging showed 
      that the RIIIJ-elicited increase in [Ca] (i) corresponded to different patterns 
      of action potentials (APs), a train of APs in L1 neurons, and sporadic firing in 
      L2 neurons. Genetically labeled mice established that L1 neurons are 
      proprioceptors. The single-cell transcriptomes of L1 and L2 neurons showed that 
      L2 neurons are Adelta-low-threshold mechanoreceptors. RIIIJ effects were replicated 
      by application of the K(v)1.1 selective antagonist, Dendrotoxin-K, in several 
      L-DRG subclasses (L1, L2, L3, and L5), suggesting the presence of functional 
      K(v)1.1/K(v)1.2 heteromeric channels. Using this approach on other neuronal 
      subclasses should ultimately accelerate the comprehensive classification and 
      characterization of individual somatosensory neuronal subclasses within a mixed 
      population.
FAU - Giacobassi, Mario J
AU  - Giacobassi MJ
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120.
FAU - Leavitt, Lee S
AU  - Leavitt LS
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120.
FAU - Raghuraman, Shrinivasan
AU  - Raghuraman S
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120.
FAU - Alluri, Rishi
AU  - Alluri R
AUID- ORCID: 0000-0002-4229-927X
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120.
FAU - Chase, Kevin
AU  - Chase K
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120.
FAU - Finol-Urdaneta, Rocio K
AU  - Finol-Urdaneta RK
AUID- ORCID: 0000-0003-2157-4532
AD  - Illawarra Health and Medical Research Institute, University of Wollongong, 
      Wollongong, NSW 2522, Australia.
FAU - Terlau, Heinrich
AU  - Terlau H
AD  - Institute of Physiology, Christian Albrechts University Kiel, Kiel 24118, 
      Germany.
FAU - Teichert, Russell W
AU  - Teichert RW
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120.
FAU - Olivera, Baldomero M
AU  - Olivera BM
AUID- ORCID: 0000-0003-4556-1410
AD  - School of Biological Sciences, University of Utah, Salt Lake City, UT 841120; 
      olivera@biology.utah.edu.
LA  - eng
GR  - P01 GM048677/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20200220
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Conotoxins)
RN  - 0 (Peptides)
RN  - 0 (Potassium Channel Blockers)
RN  - 0 (kappaM-conotoxin RIIIK, Conus radiatus)
RN  - 119128-61-9 (dendrotoxin K)
RN  - 147173-20-4 (Kv1.1 Potassium Channel)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Calcium/metabolism
MH  - Conotoxins/pharmacology
MH  - Cytosol/metabolism
MH  - Ganglia, Spinal/*cytology/drug effects
MH  - Kv1.1 Potassium Channel/antagonists & inhibitors
MH  - Mice
MH  - Mice, Transgenic
MH  - Peptides/pharmacology
MH  - Potassium Channel Blockers/pharmacology
MH  - Sensory Receptor Cells/*classification/drug effects/*physiology
MH  - Single-Cell Analysis
MH  - Transcriptome
PMC - PMC7071849
OTO - NOTNLM
OT  - DRG neurons
OT  - calcium imaging
OT  - conopeptide
OT  - neuronal subclasses
COIS- The authors declare no competing interest.
EDAT- 2020/02/23 06:00
MHDA- 2020/07/14 06:00
PMCR- 2020/08/20
CRDT- 2020/02/22 06:00
PHST- 2020/02/23 06:00 [pubmed]
PHST- 2020/07/14 06:00 [medline]
PHST- 2020/02/22 06:00 [entrez]
PHST- 2020/08/20 00:00 [pmc-release]
AID - 1911382117 [pii]
AID - 201911382 [pii]
AID - 10.1073/pnas.1911382117 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2020 Mar 10;117(10):5494-5501. doi: 
      10.1073/pnas.1911382117. Epub 2020 Feb 20.