Fluorescence Imaging of Actin Turnover Parses Early Stem Cell Lineage Divergence and Senescence

Mishra, Prakhar; Martin, D.C.; Androulakis, Ioannis P.; Moghe, Prabhas V.

doi:10.1038/s41598-019-46682-y

Cited by 19 publications

(18 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Previous confocal studies with fixed tissues showed that the actin cytoskeleton undergoes dramatic rearrangements as cells damaged by Plasmodium invasion are extruded from the midgut epithelial layer, by forming an actin ring at the base of damaged cells that is gradually constricted ( 1 ). Because traditional actin markers, such as phalloidin, are highly toxic ( 11 ) and impermeable to live cells, we decided to use the F-actin-specific SiR-actin fluorophore ( 12 ).…”

Section: Resultsmentioning

confidence: 99%

Live In Vivo Imaging of Plasmodium Invasion of the Mosquito Midgut

Trisnadi

Barillas‐Mury

2020

mSphere

View full text Add to dashboard Cite

The mosquito midgut is a critical barrier that Plasmodium parasites must overcome to complete their developmental cycle and be transmitted to a new vertebrate host. Previous confocal studies with fixed infected midguts showed that ookinetes traverse midgut epithelial cells and cause irreversible tissue damage. Here, we investigated the spatiotemporal dynamics of ookinete midgut traversal and the response of midgut cells to invasion. A novel mounting strategy was established, suitable fluorescent dye combinations were identified and protocols optimized to label mosquito tissues in vivo, and live imaging protocols using confocal microscopy were developed. Tracking data showed that ookinetes gliding on the midgut surface travel faster and farther than those that remain in the lumen or those that have invaded the epithelium. Image analysis confirmed that parasite invasion and cell traversal occur within a couple of minutes, while caspase activity in damaged cells, indicative of cellular apoptosis, and F-actin cytoskeletal rearrangements in cells extruded into the gut lumen persist for several hours. This temporal difference highlights the importance of hemocyte-mediated cellular immunity and the mosquito complement system to mount a coordinated and effective antiplasmodial response. This novel in vivo imaging protocol allowed us to continuously observe individual ookinetes in live mosquitoes within the gut lumen and during cell traversal and to capture the subsequent cellular responses to invasion in real time for several hours, without loss of tissue integrity. IMPORTANCE Malaria is one of the most devastating parasitic diseases in humans and is transmitted by anopheline mosquitoes. The mosquito midgut is a critical barrier that Plasmodium parasites must overcome to complete their developmental cycle and be transmitted to a new host. Here, we developed a new strategy to visualize Plasmodium ookinetes as they traverse the mosquito midgut and to follow the response of damaged epithelial cells by imaging live mosquitoes. Understanding the spatial and temporal aspects of these interactions is critical when developing novel strategies to disrupt disease transmission.

show abstract

Section: Resultsmentioning

confidence: 99%

Live In Vivo Imaging of Plasmodium Invasion of the Mosquito Midgut

Trisnadi

Barillas‐Mury

2020

mSphere

View full text Add to dashboard Cite

show abstract

“…Besides the abovementioned markers, new tools and approaches have been proposed to monitor MSC aging such as SiR-actin, a fluorogenic F-actin specific probe that can be used to evaluate actin turnover (Mishra et al, 2019). CyBC9 (another fluorescent probe) combined with high-throughput screening revealed accumulation of mitochondria in senescent MSCs that presumably resulted from the loss of membrane potential (Ang et al, 2019).…”

Section: Strategies For Monitoring Msc Senescencementioning

confidence: 99%

Senescence in Mesenchymal Stem Cells: Functional Alterations, Molecular Mechanisms, and Rejuvenation Strategies

Liu

Ding

Liu

et al. 2020

Front. Cell Dev. Biol.

185

136

View full text Add to dashboard Cite

Mesenchymal stem cells (MSCs) are multipotent cells capable of self-renewal and differentiation. There is increasing evidence of the therapeutic value of MSCs in various clinical situations, however, these cells gradually lose their regenerative potential with age, with a concomitant increase in cellular dysfunction. Stem cell aging and replicative exhaustion are considered as hallmarks of aging and functional attrition in organisms. MSCs do not proliferate infinitely but undergo only a limited number of population doublings before becoming senescent. This greatly hinders their clinical application, given that cultures must be expanded to obtain a sufficient number of cells for cellbased therapy. Here, we review the current knowledge of the phenotypic and functional characteristics of senescent MSCs, molecular mechanisms underlying MSCs aging, and strategies to rejuvenate senescent MSCs, which can broaden their range of therapeutic applications.

show abstract

“…Notably, various biological processes such as proliferation [10] and differentiation [11,12] are influenced by cell shape. In addition, actin cytoskeleton-mediated cell shape changes have been shown to be vital for the regulation of MSC lineage commitment [13].…”

Section: Actin Propertiesmentioning

confidence: 99%

A glance on the role of actin in osteogenic and adipogenic differentiation of mesenchymal stem cells

et al. 2020

View full text Add to dashboard Cite

Mesenchymal stem cells (MSCs) have the capacity to differentiate into multiple lineages including osteogenic and adipogenic lineages. An increasing number of studies have indicated that lineage commitment by MSCs is influenced by actin remodeling. Moreover, actin has roles in determining cell shape, nuclear shape, cell spreading, and cell stiffness, which eventually affect cell differentiation. Osteogenic differentiation is promoted in MSCs that exhibit a large spreading area, increased matrix stiffness, higher levels of actin polymerization, and higher density of stress fibers, whereas adipogenic differentiation is prevalent in MSCs with disrupted actin networks. In addition, the mechanical properties of F-actin empower cells to sense and transduce mechanical stimuli, which are also reported to influence differentiation. Various biomaterials, mechanical, and chemical interventions along with pathogeninduced actin alteration in the form of polymerization and depolymerization in MSC differentiation were studied recently. This review will cover the role of actin and its modifications through the use of different methods in inducing osteogenic and adipogenic differentiation.

show abstract

Fluorescence Imaging of Actin Turnover Parses Early Stem Cell Lineage Divergence and Senescence

Cited by 19 publications

References 46 publications

Live In Vivo Imaging of Plasmodium Invasion of the Mosquito Midgut

Live In Vivo Imaging of Plasmodium Invasion of the Mosquito Midgut

Senescence in Mesenchymal Stem Cells: Functional Alterations, Molecular Mechanisms, and Rejuvenation Strategies

A glance on the role of actin in osteogenic and adipogenic differentiation of mesenchymal stem cells

Contact Info

Product

Resources

About