Centrin-egfp
Centrin-egfp - Related Videos
11:43
Easy Measurement of Diffusion Coefficients of EGFP-tagged Plasma Membrane Proteins Using k-Space Image Correlation Spectroscopy
0 Views •
Cited by 37 •
2014
This paper provides a step by step guide to the fluctuation analysis technique k-Space Image Correlation Spectroscopy (kICS) for measuring diffusion coefficients of fluorescently labeled plasma membrane proteins in live mammalian cells.
10:03
Development of Leishmania Species Strains with Constitutive Expression of eGFP
0 Views •
2023
Here, we describe the methodology used for generating L. panamensis and L. donovani strains expressing the gene for eGFP as a stable integrated transgene using the pLEXSY system. Transfected parasites were cloned by limiting dilution, and clones with the highest fluorescence intensity in both species were selected for further use in drug screening assays.
10:20
Studying Membrane Protein Trafficking in Drosophila Photoreceptor Cells Using eGFP-Tagged Proteins
0 Views •
Cited by 1 •
2022
Here, non-invasive methods are described for localization of photoreceptor membrane proteins and assessment of retinal degeneration in the Drosophila compound eye using eGFP fluorescence.
09:53
Volumetric Imaging and Analysis of Primary Cilia in Musculoskeletal Tissue using the ARL13B-CENTRIN-2 Mouse Model
0 Views •
Cited by 1 •
2025
This method describes the exploitation of the endogenous signal in an ARL13B-CENTRIN-2 transgenic mouse model to image primary cilia in situ, in three dimensions across large areas of mineralized tissue, from tissue collection to analysis of cilia organization. This protocol may also be applied to other types of tissues.
10:54
Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression
0 Views •
Cited by 9 •
2017
In Parkinson's Disease (PD), Substantia Nigra (SNc) dopaminergic neurons degenerate, leading to motor dysfunction. Here we report a protocol for culturing ventral midbrain neurons from a mouse expressing eGFP driven by a Tyrosine Hydroxylase (TH) promoter sequence, harvesting individual fluorescent neurons from the cultures, and measuring their transcriptome using RNA-seq.
FAQs
Centrin supplies the localization information because it is associated with centrosomal structures, while EGFP provides the fluorescent readout. Together, the fusion concentrates signal in centrioles, centrosomes, and related microtubule-organizing structures rather than distributing it uniformly through the cell. This connection allows microscopy to visualize the organization of these compartments in living cells.
Centrin is a calcium-binding protein associated with centrosomal structures, so its biological identity helps direct the fusion protein to the compartments being studied. EGFP does not provide that targeting role; it supplies the detectable fluorescence. The combination therefore links a centrosome-associated protein component with an optical signal suitable for observing cellular organization and dynamics.
Time-resolved imaging can show how centrosomal structures change in position, organization, duplication, and behavior. These observations are valuable because centrosomes also organize microtubules, so their movement or reorganization can be examined alongside changes in cellular architecture. The resulting view emphasizes dynamic relationships rather than treating the centrosome as a static compartment.
Researchers express the fusion protein in living cells and use fluorescence microscopy to detect its EGFP signal at centrioles, centrosomes, and related organizing structures. They can then follow signal location and behavior over time, focusing on centrosome positioning, organization, or duplication. This workflow connects visible fluorescence patterns with changes in cellular architecture and microtubule organization.
The probe is useful when a study needs to follow centrosomal behavior during neuronal development, migration, or differentiation. Imaging can reveal where the centrosome is positioned and how its organization changes while neuronal cells undergo these processes. Such observations help investigate how centrosome-associated microtubule organization relates to the development of neuronal structure and function.
Changes in the fluorescently labeled centrosomal structures can provide information about centrosome positioning, duplication, and behavior in neuronal cells. Because these structures are related to microtubule organization, their dynamics can be considered alongside broader changes in cellular architecture. The approach therefore supports microscopy-based analysis of how intracellular organization accompanies neuronal development, migration, and differentiation.