Faglige interesser
- Autofagi
- Cellebiologi
- Membrantransport
- Nevrodegenerative sykdommer
Emneord:
Autofagi,
Cellebiologi,
Membrantransport,
Nevrodegenerative sykdommer
Publikasjoner
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Holme, Børge; Bjørnerud, Birgitte; Pedersen, Nina Marie; Rodriguez de la Ballina, Laura; Wesche, Jørgen & Haugsten, Ellen Margrethe
(2023).
Automated tracking of cell migration in phase contrast images with CellTraxx.
Scientific Reports.
ISSN 2045-2322.
13(1).
doi:
10.1038/s41598-023-50227-9.
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The ability of cells to move and migrate is required during development, but also in the adult in processes such as wound healing and immune responses. In addition, cancer cells exploit the cells’ ability to migrate and invade to spread into nearby tissue and eventually metastasize. The majority of cancer deaths are caused by metastasis and the process of cell migration is therefore intensively studied. A common way to study cell migration is to observe cells through an optical microscope and record their movements over time. However, segmenting and tracking moving cells in phase contrast time-lapse video sequences is a challenging task. Several tools to track the velocity of migrating cells have been developed. Unfortunately, most of the automated tools are made for fluorescence images even though unlabelled cells are often preferred to avoid phototoxicity. Consequently, researchers are constrained with laborious manual tracking tools using ImageJ or similar software. We have therefore developed a freely available, user-friendly, automated tracking tool called CellTraxx. This software makes it easy to measure the velocity and directness of migrating cells in phase contrast images. Here, we demonstrate that our tool efficiently recognizes and tracks unlabelled cells of different morphologies and sizes (HeLa, RPE1, MDA-MB-231, HT1080, U2OS, PC-3) in several types of cell migration assays (random migration, wound healing and cells embedded in collagen). We also provide a detailed protocol and download instructions for CellTraxx.
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Munson, Michael Joseph; Mathai, Benan J.; Ng, Matthew; Trachsel Moncho, Laura Cristina; de la Ballina, Laura R & Schultz, Sebastian W.
[Vis alle 13 forfattere av denne artikkelen]
(2021).
GAK and PRKCD are positive regulators of PRKN-independent mitophagy.
Nature Communications.
ISSN 2041-1723.
12(1).
doi:
10.1038/s41467-021-26331-7.
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The mechanisms involved in programmed or damage-induced removal of mitochondria by mitophagy remains elusive. Here, we have screened for regulators of PRKN-independent mitophagy using an siRNA library targeting 197 proteins containing lipid interacting domains. We identify Cyclin G-associated kinase (GAK) and Protein Kinase C Delta (PRKCD) as regulators of PRKN-independent mitophagy, with both being dispensable for PRKN- dependent mitophagy and starvation-induced autophagy. We demonstrate that the kinase activity of both GAK and PRKCD are required for efficient mitophagy in vitro, that PRKCD is present on mitochondria, and that PRKCD facilitates recruitment of ULK1/ATG13 to early autophagic structures. Importantly, we demonstrate in vivo relevance for both kinases in the regulation of basal mitophagy. Knockdown of GAK homologue (gakh-1) in C. elegans or knockout of PRKCD homologues in zebrafish led to significant inhibition of basal mitophagy, highlighting the evolutionary relevance of these kinases in mitophagy regulation.
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Se alle arbeider i Cristin
Publisert
3. jan. 2017 18:15
- Sist endret
17. aug. 2023 13:53