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Sequential membrane remodeling by cholesterol distinctly modulates HCN channels in naïve and neuropathic DRG neurons.

Overview

  1. Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, MO, USA
  2. Department of Pharmacology and Physiology, Saint Louis University School of Medicine, St. Louis, MO, USA
Institutions: Saint Louis University (United States)
Journal: The Journal of general physiology, volume 158, issue 3, article e202513925
Dates: received 4 November 2025; accepted 19 February 2026; published online 12 March 2026; in print May 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.1085/jgp.202513925 · PMID 41818135 · PMCID PMC12981344 · OpenAlex W7135207916
Open access: hybrid, a free copy (OpenAlex)
Status: data only
Categories: rat (organism), pain (population), cellular / molecular (subfield)
Methods: Statistics, Evoked potentials, Single-unit activity, calcium imaging
MeSH: Cell Membrane*, Cholesterol*, Ganglia, Spinal*, Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels*, Neurons*, Animals, Ion Channel Gating, Male, Nociceptors, Rats, Rats, Sprague-Dawley (* major topic)
Topic: Pain Mechanisms and Treatments (Physiology, Medicine), according to OpenAlex
Funding: NINDS NIH HHS (R01NS119263, R01 NS119263); NIGMS NIH HHS (R35 GM154778, R35GM154778); SLU Institute for Drug & Biotherapeutic Innovation; National Institute of Neurological Disorders and Stroke (R01NS119263); National Institute of General Medical Sciences (R35GM154778); Edward A. Doisy Department of Biochemistry and Molecular Biology at Saint Louis University School of Medicine (Doisy Fund); Department of Defense Chronic Pain Management Research Program (#CP220080P1); Department of Pharmacology and Physiology, Saint Louis University
Citations: cited by 3 papers (Europe PMC); 73 references in the paper
Research resources: The tsA-201 cell line RRID:CVCL_2737, RRID:RGD_70508, RRID:SCR_000034

Abstract

Cholesterol, abundantly present in distinct plasma membrane pools, is a critical modulator of ion channel function, including hyperpolarization-activated cyclic nucleotide-gated (HCN) channels that regulate the excitability of dorsal root ganglion (DRG) nociceptor neurons. Depletion of membrane cholesterol potentiated HCN channel opening and accelerated activation kinetics, whereas cholesterol supplementation reduced channel opening and slowed activation kinetics. However, the relative contributions of cholesterol that organizes ordered membrane domains (OMDs) versus freely accessible cholesterol pools to HCN channel modulation remain unknown. Using fluorescence lifetime imaging microscopy, FRET and fluorescence anisotropy techniques, we examined how supplementing cholesterol alters plasma membrane properties and HCN gating in nociceptor DRG neurons. We uncovered a process of sequential, stepwise membrane remodeling: an initial phase with OMD expansion and a rapid rise in free cholesterol, followed by continued accumulation of free cholesterol without further OMD expansion. Notably, the slope factor of the HCN G-V relationship is sensitive to OMD expansion but remains unaffected by changes in free cholesterol. Other gating parameters, including open probability and activation kinetics, were affected by elevating free cholesterol. In a rat model of nerve injury, where DRG neurons exhibit reduced free cholesterol levels and smaller OMDs, HCN channel modulation by cholesterol involves contributions from both OMD expansion and free cholesterol accumulation. In contrast, in naïve DRG neurons—characterized by high cholesterol and large OMDs—modulation occurs mostly via increased free cholesterol. These findings provide mechanistic insights into cholesterol-dependent modulation of ion channels and its role in neuropathic pain.

Reproduced under the paper's license (CC BY), from the paper cited above.

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Data

Datasets cited

Data availability

All source data in Figs. 1, 2, 3, 4, 5, and 6 are provided as a Source Data file in figshare.com as https://doi.org/10.6084/m9.figshare.29983183, and additional data are in the online supplemental material.

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

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Version 1, 30 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 6 authors, 11 MeSH terms, 8 funders, 73 references, 3 RRIDs.

Cite

This paper

Handlin, L. J., Gieré, C., Dumaire, N. L., Salih, L., Moutal, A., & Dai, G. (2026). Sequential membrane remodeling by cholesterol distinctly modulates HCN channels in naïve and neuropathic DRG neurons. The Journal of general physiology, 158(3), e202513925. https://doi.org/10.1085/jgp.202513925

BibTeX

@article{handlin2026sequential,
author = {Handlin, Lucas J and Gieré, Clémence and Dumaire, Nicolas LA and Salih, Lyuba and Moutal, Aubin and Dai, Gucan},
title = {{Sequential membrane remodeling by cholesterol distinctly modulates HCN channels in naïve and neuropathic DRG neurons}},
journal = {The Journal of general physiology},
year = {2026},
month = mar,
volume = {158},
number = {3},
pages = {e202513925},
publisher = {Rockefeller University Press},
issn = {0022-1295},
doi = {10.1085/jgp.202513925},
url = {https://doi.org/10.1085/jgp.202513925},
pmid = {41818135},
pmcid = {PMC12981344}
}

RIS

TY - JOUR
AU - Handlin, Lucas J
AU - Gieré, Clémence
AU - Dumaire, Nicolas LA
AU - Salih, Lyuba
AU - Moutal, Aubin
AU - Dai, Gucan
TI - Sequential membrane remodeling by cholesterol distinctly modulates HCN channels in naïve and neuropathic DRG neurons
T2 - The Journal of general physiology
J2 - J Gen Physiol
PY - 2026
DA - 2026/03/12
VL - 158
IS - 3
SP - e202513925
SN - 0022-1295
PB - Rockefeller University Press
DO - 10.1085/jgp.202513925
UR - https://doi.org/10.1085/jgp.202513925
LA - en
ER -

CSL-JSON

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