Cell membranes consist of heterogeneous
lipid domains that influence key cellular processes, including signal transduction, endocytosis, and electrical excitability. The goal of this study was to assess the size of
cholesterol-enriched ordered membrane domains (OMD) in various cell types. Multiple cell types were tested using fluorescence lifetime imaging microscopy (FLIM) and Förster resonance energy transfer (FRET), whereby small nociceptor DRG neurons and cardiac pacemaker cells displayed the highest FRET intensities. This implies that electrically active cells tend to have large OMDs. Treatment of cells with the
cholesterol-extracting
reagent β-
cyclodextrin (β-CD) led to a decrease in FRET, indicating a reduction in the OMD size, whereas
detergents known to promote domain coalescence in
artificial membranes increased OMD size. In an in vitro
fatty liver model,
palmitate supplementation increased FRET whereas
oleate supplementation decreased FRET in isolated primary murine hepatocytes, highlighting the importance of unsaturated
lipid tails in
lipid domain organization. Disruption of OMD using β-CD potentiated action potential firing in nociceptor DRG neurons and decreased the free energy needed for opening native hyperpolarization-activated
cyclic nucleotide-gated (HCN) channels. After disrupting the OMD, HCN channels exhibited an increased relative open probability at the resting membrane potential (RMP). A significant reduction in FRET was observed in both a
chemotherapy-induced
neuropathic pain model and a spared nerve injury model of
neuropathic pain, consistent with disrupted or shrunken OMD in these models. Collectively, these findings show that disturbances in
lipid domains may contribute to the progression of
neuropathic pain, and they suggest new therapeutic strategies to achieve
pain relief.