Spatiotemporal dynamics of microRNA during epithelial collective cell migration

Dean, Zachary; Riahi, Reza; Wong, Pak Kin

doi:10.1016/j.biomaterials.2014.10.022

Cited by 23 publications

(28 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To investigate leader cells, we applied a double-stranded locked nucleic acid (dsLNA) assay for detecting gene expression at the single cell level 17 , 27 , 28 and compared the acquired gene expression with other leader cell characteristics. Leader cells are typically characterized by their phenotypes, including enlarged morphology and active lamellipodium on the migration tip.…”

Section: Resultsmentioning

confidence: 99%

Notch1–Dll4 signalling and mechanical force regulate leader cell formation during collective cell migration

et al. 2015

Self Cite

View full text Add to dashboard Cite

At the onset of collective cell migration, a subset of cells within an initially homogenous population acquires a distinct “leader” phenotype with characteristic morphology and motility. However, the factors driving leader cell formation as well as the mechanisms regulating leader cell density during the migration process remain to be determined. Here, we use single cell gene expression analysis and computational modeling to show that leader cell identity is dynamically regulated by Dll4 signaling through both Notch1 and cellular stress in a migrating epithelium. Time-lapse microscopy reveals that Dll4 is induced in leader cells after the creation of the cell-free region and leader cells are regulated via Notch1-Dll4 lateral inhibition. Furthermore, mechanical stress inhibits Dll4 expression and leader cell formation in the monolayer. Collectively, our findings suggest that a reduction of mechanical force near the boundary promotes Notch1-Dll4 signaling to dynamically regulate the density of leader cells during collective cell migration.

show abstract

Section: Resultsmentioning

confidence: 99%

Notch1–Dll4 signalling and mechanical force regulate leader cell formation during collective cell migration

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…The LNA probe was labeled with a fluorophore (6-FAM) at the 5’ end for fluorescence detection. The design process of LNA probes was reported previously 25, 26 . Briefly, the LNA probe was designed to be complementary to the loop region of the target mRNA structure.…”

Section: Methodsmentioning

confidence: 99%

A nanobiosensor for dynamic single cell analysis during microvascular self-organization

et al. 2016

View full text Add to dashboard Cite

The formation of microvascular networks plays essential roles in regenerative medicine and tissue engineering. Nevertheless, the self-organization mechanisms underlying the dynamic morphogenic process are poorly understood due to a paucity of effective tools for mapping the spatiotemporal dynamics of single cell behaviors. By establishing a single cell nanobiosensor along with live cell imaging, we perform dynamic single cell analysis of the morphology, displacement, and gene expression during microvascular self-organization. Dynamic single cell analysis reveals that endothelial cells self-organize into subpopulations with specialized phenotypes to form microvascular networks and identifies the involvement of Notch1-Dll4 signaling in regulating the cell subpopulations. The cell phenotype correlates with the initial Dll4 mRNA expression level and each subpopulation displays a unique dynamic Dll4 mRNA expression profile. Pharmacological perturbations and RNA interference of Notch1-Dll4 signaling modulate the cell subpopulations and modify the morphology of the microvascular network. Taken together, the nanobiosensor enables a dynamic single cell analysis approach underscoring the importance of Notch1-Dll4 signaling in microvascular self-organization.

show abstract

“…A probe-modified technique can improve the hybridization efficiency and detection sensitivity. Kin Wong et al [42] have reported an intracellular miRNAs detection strategy using a doublestranded locked nucleic acid (dsLNA) probe, which showed high performance in detecting the dynamic regulation and dose-dependent modulation of miRNAs.…”

Section: Mirna Imaging Analysismentioning

confidence: 99%

Intracellular and Organic miRNA In Situ Detection

Zhang

Dong

Tian

2015

SpringerBriefs in Molecular Science

View full text Add to dashboard Cite

The detection methods of miRNA in intracellular or organisms fall into two broad categories: indirect detection and direct analysis. The indirect measurement of the expression levels of miRNAs in cells and tissues involves cells lysis and detection by qRT-PCR, northern blotting, or microarray hybridization. The direct analysis methods are a noninvasive manner for repetitively monitoring and obtaining real-time imaging of the intracellular miRNA by using imaging analysis or in situ hybridization (ISH). Technologies for direct detection of the temporal and spatial expression sequence of specific miRNA in cells or tissues are extremely important for elucidating miRNA biology. The progress of optical imaging techniques with multimodal reporter systems holds great promise for noninvasive and real-time imaging of molecular agent expression in living cell. Recent progress in nanotechnology and imaging detection techniques leads to multifunctional nanoprobe with specific-transfection, tracing, and regulation function in intracellular miRNA detection. ISH holds great promise for visualization of the spatial localization of RNA at the tissue, cellular, and even subcellular level.Keywords Intracellular miRNA · Organic miRNA · In situ detection · Cell imaging analysis · Functional nanoprobeThe detection of intracellular miRNA is significant in the development of gene therapy and gene medicines. The deficiency of small size and degradation of the mature miRNAs make it difficult to directly transfer the specific miRNA into the cell. A noninvasive manner for repetitively monitoring and obtaining real-time imaging of the intracellular miRNA is required for the analysis of miRNAs in practical clinic application. Efficient gene vectors, including viral [1] and nonviral categories, [2] were usually required to translocate miRNA probe through membrane barrier into the cell. Retrovirus and adenovirus vectors can be effectively transferred via the gene probes (DNA, RNA) into most cell lines. However, the

show abstract

Spatiotemporal dynamics of microRNA during epithelial collective cell migration

Cited by 23 publications

References 37 publications

Notch1–Dll4 signalling and mechanical force regulate leader cell formation during collective cell migration

Notch1–Dll4 signalling and mechanical force regulate leader cell formation during collective cell migration

A nanobiosensor for dynamic single cell analysis during microvascular self-organization

Intracellular and Organic miRNA In Situ Detection

Contact Info

Product

Resources

About