Time Series Analyzer

Goulding, Gunilla

doi:10.2175/193864702784900156

Cited by 2 publications

(1 citation statement)

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“…The threshold was then adjusted to select for fluorescent events within the difference images stack, and the inbuilt ‘Analyze Particles’ function was employed to select and analyse fluorescent events above the set threshold. The fluorescence profile over‐time of every event highlighted by the Analyze Particles function was assessed with the Time Series Analyzer V3 Plugin (Balaji, 2014). Events with minimal change in fluorescence, an irregular fluorescence profile and/or lateral movement of the fluorescence signal were excluded from the analysis.…”

Section: Methodsmentioning

confidence: 99%

TRPC3 signalling contributes to the biogenesis of extracellular vesicles

Padbury,

Bálint,

Carollo

et al. 2023

J of Extracellular Bio

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Extracellular vesicles (EVs) contribute to a wide range of pathological processes including cancer progression, yet the molecular mechanisms underlying their biogenesis remain incompletely characterized. The development of tetraspanin‐based pHluorin reporters has enabled the real‐time analysis of EV release at the plasma membrane. Here, we employed CD81‐pHluorin to investigate mechanisms of EV release in ovarian cancer (OC) cells and report a novel role for the Ca2+‐permeable transient receptor potential (TRP) channel TRPC3 in EV‐mediated communication. We found that specific activation of TRPC3 increased Ca2+ signalling in SKOV3 cells and stimulated an immediate increase in EV release. Ca2+‐stimulants histamine and ionomycin likewise induced EV release, and imaging analysis revealed distinct stimulation‐dependent temporal and spatial release dynamics. Interestingly, inhibition of TRPC3 attenuated histamine‐stimulated Ca2+‐entry and EV release, indicating that TRPC3 is likely to act downstream of histamine signalling in EV biogenesis. Furthermore, we found that direct activation of TRPC3 as well as the application of EVs derived from TRPC3‐activated cells increased SKOV3 proliferation. Our data provides insights into the molecular mechanisms and dynamics underlying EV release in OC cells, proposing a key role for TRPC3 in EV biogenesis.

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Section: Methodsmentioning

confidence: 99%