Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

Ashdown, George W.; Pandžić, Elvis; Cope, A. D.; Wiseman, Paul W.; Owen, Dylan M.

doi:10.3791/53749-v

Cited by 4 publications

(1 citation statement)

References 9 publications

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“…Background noise was removed using the subtract background function in F iji . To analyze the actin filament flow, flow velocity vectors ( v x , v y ) were obtained from spatiotemporal image correlation spectroscopy analysis using M atlab scripts (Ashdown et al ., 2015) with parameter settings: pixelSize 0.146, timeframe 5, tauLimit 4, filtering FourierWhole, MoveAverage 21, ROIsize 16, ROIshift 4, TOIsize 3, and TOIshift 1 as described in the manual (Ashdown et al ., 2015). After the flow velocity vectors were acquired from each cell, all angles of the flow velocity vectors from the entire analyzed frames were calculated and normalized as probability to get the direction distribution of the actin filament flow.…”

Section: Methodsmentioning

confidence: 99%

Basally distributed actin array drives embryonic hypocotyl elongation during the seed‐to‐seedling transition in Arabidopsis

Cui

Zou

2023

New Phytologist

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Summary Seed germination is a vital developmental transition for the production of progeny by sexual reproduction in spermatophytes. The seed‐to‐seedling transition is predominately driven by hypocotyl cell elongation. However, the mechanism that underlies hypocotyl growth remains largely unknown. In this study, we characterized the actin array reorganization in embryonic hypocotyl epidermal cells. Live‐cell imaging revealed a basally organized actin array formed during hypocotyl cell elongation. This polarized actin assembly is a barrel‐shaped network, which comprises a backbone of longitudinally aligned actin cables and a fine actin cap linking these cables. We provide genetic evidence that the basal actin array formation requires formin‐mediated actin polymerization and directional movement of actin filaments powered by myosin XIs. In fh1‐1 and xi3ko mutants, actin filaments failed to reorganize into the basal actin array, and the hypocotyl cell elongation was inhibited compared with wild‐type plants. Collectively, our work uncovers the molecular mechanisms for basal actin array assembly and demonstrates the connection between actin polarization and hypocotyl elongation during seed‐to‐seedling transition.

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Section: Methodsmentioning

confidence: 99%

Basally distributed actin array drives embryonic hypocotyl elongation during the seed‐to‐seedling transition in Arabidopsis

Cui

Zou

2023

New Phytologist

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HIV-1 diverts cortical actin for particle assembly and release

Dibsy,

Bremaud,

Mak

et al. 2023

Nat Commun

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Enveloped viruses assemble and bud from the host cell membranes. Any role of cortical actin in these processes have often been a source of debate. Here, we assessed if cortical actin was involved in HIV-1 assembly in infected CD4 T lymphocytes. Our results show that preventing actin branching not only increases HIV-1 particle release but also the number of individual HIV-1 Gag assembly clusters at the T cell plasma membrane. Indeed, in infected T lymphocytes and in in vitro quantitative model systems, we show that HIV-1 Gag protein prefers areas deficient in F-actin for assembling. Finally, we found that the host factor Arpin, an inhibitor of Arp2/3 branched actin, is recruited at the membrane of infected T cells and it can associate with the viral Gag protein. Altogether, our data show that, for virus assembly and particle release, HIV-1 prefers low density of cortical actin and may favor local actin debranching by subverting Arpin.

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Current capabilities and future perspectives of FCS: super-resolution microscopy, machine learning, and in vivo applications

Sankaran

Wohland

2023

Commun Biol

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Fluorescence correlation spectroscopy (FCS) is a single molecule sensitive tool for the quantitative measurement of biomolecular dynamics and interactions. Improvements in biology, computation, and detection technology enable real-time FCS experiments with multiplexed detection even in vivo. These new imaging modalities of FCS generate data at the rate of hundreds of MB/s requiring efficient data processing tools to extract information. Here, we briefly review FCS’s capabilities and limitations before discussing recent directions that address these limitations with a focus on imaging modalities of FCS, their combinations with super-resolution microscopy, new evaluation strategies, especially machine learning, and applications in vivo.

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Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data

Cited by 4 publications

References 9 publications

Basally distributed actin array drives embryonic hypocotyl elongation during the seed‐to‐seedling transition in Arabidopsis

Basally distributed actin array drives embryonic hypocotyl elongation during the seed‐to‐seedling transition in Arabidopsis

HIV-1 diverts cortical actin for particle assembly and release

Current capabilities and future perspectives of FCS: super-resolution microscopy, machine learning, and in vivo applications

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