Intracellular potassium depletion enhances apoptosis induced by staurosporine in cultured trigeminal satellite glial cells

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dc.contributor.authorBustamante, Hedie A.en
dc.contributor.authorEhrich, Marion F.en
dc.contributor.authorKlein, Bradley G.en
dc.contributor.departmentBiomedical Sciences and Pathobiologyen
dc.date.accessioned2021-07-21T17:46:10Zen
dc.date.available2021-07-21T17:46:10Zen
dc.date.issued2021-06-30en
dc.description.abstractPurpose Satellite glial cells (SGC) surrounding neurons in sensory ganglia can buffer extracellular potassium, regulating the excitability of injured neurons and possibly influencing a shift from acute to neuropathic pain. SGC apoptosis may be a key component in this process. This work evaluated induction or enhancement of apoptosis in cultured trigeminal SGC following changes in intracellular potassium [K]ic. Materials and methods We developed SGC primary cultures from rat trigeminal ganglia (TG). Purity of our cultures was confirmed using immunofluorescence and western blot analysis for the presence of the specific marker of SGC, glutamine synthetase (GS). SGC [K]ic was depleted using hypo-osmotic shock and 4 mM bumetanide plus 10 mM ouabain. [K]ic was measured using the K+ fluorescent indicator potassium benzofuran isophthalate (PBFI-AM). Results SGC tested positive for GS and hypo-osmotic shock induced a significant decrease in [K]ic at every evaluated time. Cells were then incubated for 5 h with either 2 mM staurosporine (STS) or 20 ng/ml of TNF-alpha and evaluated for early apoptosis and late apoptosis/necrosis by flow cytometry using annexin V and propidium iodide. A significant increase in early apoptosis, from 16 to 38%, was detected in SGC with depleted [K]ic after incubation with STS. In contrast, TNF-alpha did not increase early apoptosis in normal or [K]ic depleted SGC. Conclusion Hypo-osmotic shock induced a decrease in intracellular potassium in cultured trigeminal SGC and this enhanced apoptosis induced by STS that is associated with the mitochondrial pathway. These results suggest that K+ dysregulation may underlie apoptosis in trigeminal SGC.en
dc.description.notesThis work was supported by the Virginia-Maryland College of Veterinary Medicine under Internal Research Competition (IRC) [Grant 4-41588].en
dc.description.sponsorshipVirginia-Maryland College of Veterinary Medicine under Internal Research Competition (IRC) [4-41588]en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1080/08990220.2021.1941843en
dc.identifier.eissn1369-1651en
dc.identifier.issn0899-0220en
dc.identifier.pmid34187291en
dc.identifier.urihttp://hdl.handle.net/10919/104245en
dc.language.isoenen
dc.rightsCreative Commons Attribution-NonCommercial-NoDerivs 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectSatellite glial cellsen
dc.subjectApoptosisen
dc.subjectstaurosporineen
dc.subjectneuropathic painen
dc.titleIntracellular potassium depletion enhances apoptosis induced by staurosporine in cultured trigeminal satellite glial cellsen
dc.title.serialSomatosensory and Motor Researchen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.dcmitypeStillImageen

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