scholarly journals The differential interaction of snRNPs with pre-mRNA reveals splicing kinetics in living cells

2010 ◽  
Vol 191 (1) ◽  
pp. 75-86 ◽  
Author(s):  
Martina Huranová ◽  
Ivan Ivani ◽  
Aleš Benda ◽  
Ina Poser ◽  
Yehuda Brody ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Comprehensive Analysis ◽  
Mrna Splicing ◽  
Differential Interaction ◽  
Time Period ◽  
Rnp Complex ◽  
Core Components

Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleoprotein (RNP) complex composed of five small nuclear RNP particles (snRNPs) and additional proteins. Using live cell imaging of GFP-tagged snRNP components expressed at endogenous levels, we examined how the spliceosome assembles in vivo. A comprehensive analysis of snRNP dynamics in the cell nucleus enabled us to determine snRNP diffusion throughout the nucleoplasm as well as the interaction rates of individual snRNPs with pre-mRNA. Core components of the spliceosome, U2 and U5 snRNPs, associated with pre-mRNA for 15–30 s, indicating that splicing is accomplished within this time period. Additionally, binding of U1 and U4/U6 snRNPs with pre-mRNA occurred within seconds, indicating that the interaction of individual snRNPs with pre-mRNA is distinct. These results are consistent with the predictions of the step-wise model of spliceosome assembly and provide an estimate on the rate of splicing in human cells.

scholarly journals A Decade of Imaging Cellular Motility and Interaction Dynamics in the Immune System

Science ◽  
2012 ◽  
Vol 336 (6089) ◽  
pp. 1676-1681 ◽  
Author(s):  
Ronald N. Germain ◽  
Ellen A. Robey ◽  
Michael D. Cahalan
Keyword(s):  
Lymph Nodes ◽  
Live Cell Imaging ◽  
Quantitative Measurement ◽  
Cell Imaging ◽  
Plasma Cells ◽  
Effector T Cells ◽  
Cellular Interactions ◽  
Cellular Motility ◽  
Response Dynamics

To mount an immune response, lymphocytes must recirculate between the blood and lymph nodes, recognize antigens upon contact with specialized presenting cells, proliferate to expand a small number of clonally relevant lymphocytes, differentiate to antibody-producing plasma cells or effector T cells, exit from lymph nodes, migrate to tissues, and engage in host-protective activities. All of these processes involve motility and cellular interactions—events that were hidden from view until recently. Introduced to immunology by three papers in this journal in 2002, in vivo live-cell imaging studies are revealing the behavior of cells mediating adaptive and innate immunity in diverse tissue environments, providing quantitative measurement of cellular motility, interactions, and response dynamics. Here, we review themes emerging from such studies and speculate on the future of immunoimaging.

A new visible-light-excitable ICT-CHEF-mediated fluorescence ‘turn-on’ probe for the selective detection of Cd2+ in a mixed aqueous system with live-cell imaging

2015 ◽  
Vol 44 (12) ◽  
pp. 5763-5770 ◽  
Author(s):  
Shyamaprosad Goswami ◽  
Krishnendu Aich ◽  
Sangita Das ◽  
Chitrangada Das Mukhopadhyay ◽  
Deblina Sarkar ◽  
...  
Keyword(s):  
Visible Light ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Aqueous System ◽  
Live Cell ◽  
Selective Detection ◽  
Turn On ◽  
Fluorescence Turn On

A new quinoline based sensor was developed and applied for the selective detection of Cd2+ both in vitro and in vivo.

scholarly journals Regeneration of the neurogliovascular unit visualized in vivo by transcranial live-cell imaging

2020 ◽  
Vol 343 ◽  
pp. 108808 ◽  
Author(s):  
Margarita Arango-Lievano ◽  
Yann Dromard ◽  
Pierre Fontanaud ◽  
Chrystel Lafont ◽  
Patrice Mollard ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell

An aggregation-induced phosphorescence probe for calcium ion-specific detection and live-cell imaging in Arabidopsis thaliana

2019 ◽  
Vol 55 (33) ◽  
pp. 4841-4844 ◽  
Author(s):  
Guilin Chen ◽  
Zaicai Zhou ◽  
Hui Feng ◽  
Chenyan Zhang ◽  
Yifan Wang ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Calcium Ion ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Molecular Probe ◽  
Specific Detection ◽  
Phosphorescence Probe

A molecular probe with aggregation-induced phosphorescence (AIP) properties for calcium ion-specific detection and imaging in vivo was designed.

scholarly journals Detection and Characterization of Protein Interactions In Vivo by a Simple Live-Cell Imaging Method

PLoS ONE ◽  
2013 ◽  
Vol 8 (5) ◽  
pp. e62195 ◽  
Author(s):  
Oriol Gallego ◽  
Tanja Specht ◽  
Thorsten Brach ◽  
Arun Kumar ◽  
Anne-Claude Gavin ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Imaging Method

scholarly journals Live Cell Imaging of Butterfly Pupal and Larval Wings In Vivo

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0128332 ◽  
Author(s):  
Yoshikazu Ohno ◽  
Joji M. Otaki
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell

scholarly journals PinMol: Python application for designing molecular beacons for live cell imaging of endogenous mRNAs

2018 ◽  
Author(s):  
Livia V. Bayer ◽  
Omar S. Omar ◽  
Diana P. Bratu ◽  
Irina E. Catrina
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Molecular Beacon ◽  
Structural Information ◽  
Live Cell ◽  
Molecular Beacons ◽  
Intramolecular Interactions ◽  
Live Cells ◽  
Target Rna

ABSTRACTMolecular beacons are nucleic acid oligomers labeled with a fluorophore and a quencher that fold in a hairpin-shaped structure, which fluoresce only when bound to their target RNA. They are used for the visualization of endogenous mRNAs in live cells. Here, we report a Python program (PinMol) that designs molecular beacons best suited for live cell imaging by using structural information from secondary structures of the target RNA, predicted via energy minimization approaches. PinMol takes into account the accessibility of the targeted regions, as well as the inter- and intramolecular interactions of each selected probe. To demonstrate its applicability, we synthesized an oskar mRNA-specific molecular beacon (osk1236), which is selected by PinMol to target a more accessible region than a manually designed oskar-specific molecular beacon (osk2216). We previously demonstrated osk2216 to be efficient in detecting oskar mRNA in in vivo experiments. Here, we show that osk1236 outperformed osk2216 in live cell imaging experiments.

scholarly journals Direct observation of aggregate-triggered selective autophagy

2021 ◽  
Author(s):  
Anne FJ Janssen ◽  
Giel Korsten ◽  
Wilco Nijenhuis ◽  
Eugene Katrukha ◽  
Lukas Kapitein
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Relative Timing ◽  
Imaging Studies ◽  
Selective Autophagy ◽  
The Core ◽  
Spatiotemporal Regulation ◽  
Core Components ◽  
Inducible Protein

Degradation of aggregates by selective autophagy is important as damaged proteins may impose a threat to cellular homeostasis. Although the core components of the autophagy machinery are well-characterized, the spatiotemporal regulation of many selective autophagy processes, including aggrephagy, remains largely unexplored. Furthermore, because most live-cell imaging studies have so far focused on starvation-induced autophagy, little is known about the dynamics of aggrephagy. Here, we describe the development and application of the mKeima-PIM assay, which enables live-cell observation of autophagic turnover and degradation of inducible protein aggregates in conjunction with key autophagy players. This allowed us to quantify the relative timing and duration of different steps of aggrephagy and revealed the short-lived nature of the autophagosome. The assay furthermore showed the spatial distribution of omegasome formation, highlighting that autophagy initiation is directly instructed by the cargo. Moreover, we found that nascent autophagosomes mostly remain immobile until acidification occurs. Thus, our assay provides new insights into the spatiotemporal regulation and dynamics of aggrephagy.

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