Combined Immunochemistry and Live Imaging of Fluorescent Protein Expressing Neurons in Mouse Brain

2015 ◽  
pp. 357-373
Author(s):  
Ruth M. Empson ◽  
Malinda L. S. Tantirigama ◽  
Manfred J. Oswald ◽  
Stephanie M. Hughes ◽  
Thomas Knöpfel
Keyword(s):  
Mouse Brain ◽  
Fluorescent Protein ◽  
Live Imaging

Illuminating subcellular structures and dynamics in plants: a fluorescent protein toolboxThis review is one of a selection of papers published in the Special Issue on Plant Cell Biology.

2006 ◽  
Vol 84 (4) ◽  
pp. 515-522 ◽  
Author(s):  
Preetinder K. Dhanoa ◽  
Alison M. Sinclair ◽  
Robert T. Mullen ◽  
Jaideep Mathur
Keyword(s):  
Plant Cell ◽  
Cell Biology ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Live Imaging ◽  
Living Cells ◽  
Special Issue ◽  
Exciting Possibility ◽  
Selection Of

The discovery and development of multicoloured fluorescent proteins has led to the exciting possibility of observing a remarkable array of subcellular structures and dynamics in living cells. This minireview highlights a number of the more common fluorescent protein probes in plants and is a testimonial to the fact that the plant cell has not lagged behind during the live-imaging revolution and is ready for even more in-depth exploration.

Application of in utero electroporation and live imaging in the analyses of neuronal migration during mouse brain development

2012 ◽  
Vol 45 (1) ◽  
pp. 1-6 ◽  
Author(s):  
Yoshiaki V. Nishimura ◽  
Tomoyasu Shinoda ◽  
Yutaka Inaguma ◽  
Hidenori Ito ◽  
Koh-ichi Nagata
Keyword(s):  
Brain Development ◽  
Mouse Brain ◽  
Neuronal Migration ◽  
In Utero ◽  
Live Imaging ◽  
In Utero Electroporation ◽  
Mouse Brain Development

scholarly journals LIM Kinase 1 and Cofilin Regulate Actin Filament Population Required for Dynamin-dependent Apical Carrier Fission from the Trans-Golgi Network

2009 ◽  
Vol 20 (1) ◽  
pp. 438-451 ◽  
Author(s):  
Susana B. Salvarezza ◽  
Sylvie Deborde ◽  
Ryan Schreiner ◽  
Fabien Campagne ◽  
Michael M. Kessels ◽  
...  
Keyword(s):  
Actin Filaments ◽  
Fluorescent Protein ◽  
Live Imaging ◽  
Actin Dynamics ◽  
Lim Kinase ◽  
Trans Golgi Network ◽  
Lim Kinase 1 ◽  
Photoactivatable Gfp ◽  
Green Fluorescent ◽  
Golgi Network

The functions of the actin cytoskeleton in post-Golgi trafficking are still poorly understood. Here, we report the role of LIM Kinase 1 (LIMK1) and its substrate cofilin in the trafficking of apical and basolateral proteins in Madin-Darby canine kidney cells. Our data indicate that LIMK1 and cofilin organize a specialized population of actin filaments at the Golgi complex that is selectively required for the emergence of an apical cargo route to the plasma membrane (PM). Quantitative pulse-chase live imaging experiments showed that overexpression of kinase-dead LIMK1 (LIMK1-KD), or of LIMK1 small interfering RNA, or of an activated cofilin mutant (cofilin S3A), selectively slowed down the exit from the trans-Golgi network (TGN) of the apical PM marker p75-green fluorescent protein (GFP) but did not interfere with the apical PM marker glycosyl phosphatidylinositol-YFP or the basolateral PM marker neural cell adhesion molecule-GFP. High-resolution live imaging experiments of carrier formation and release by the TGN and analysis of peri-Golgi actin dynamics using photoactivatable GFP suggest a scenario in which TGN-localized LIMK1-cofilin regulate a population of actin filaments required for dynamin-syndapin-cortactin–dependent generation and/or fission of precursors to p75 transporters.

Distribution of corticotropin-releasing factor neurons in the mouse brain: a study using corticotropin-releasing factor-modified yellow fluorescent protein knock-in mouse

2016 ◽  
Vol 222 (4) ◽  
pp. 1705-1732 ◽  
Author(s):  
Junko Kono ◽  
Kohtarou Konno ◽  
Ashraf Hossain Talukder ◽  
Toshimitsu Fuse ◽  
Manabu Abe ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Corticotropin Releasing Factor

scholarly journals Live Imaging of Drosophila Brain Neuroblasts Reveals a Role for Lis1/Dynactin in Spindle Assembly and Mitotic Checkpoint Control

2005 ◽  
Vol 16 (11) ◽  
pp. 5127-5140 ◽  
Author(s):  
Karsten H. Siller ◽  
Madeline Serr ◽  
Ruth Steward ◽  
Tom S. Hays ◽  
Chris Q. Doe
Keyword(s):  
Cell Cycle ◽  
Fluorescent Protein ◽  
Spindle Assembly ◽  
Mitotic Checkpoint ◽  
Live Imaging ◽  
Cell Cycle Checkpoint ◽  
Checkpoint Control ◽  
Larval Brain ◽  
Independent Functions

Lis1 is required for nuclear migration in fungi, cell cycle progression in mammals, and the formation of a folded cerebral cortex in humans. Lis1 binds dynactin and the dynein motor complex, but the role of Lis1 in many dynein/dynactin-dependent processes is not clearly understood. Here we generate and/or characterize mutants for Drosophila Lis1 and a dynactin subunit, Glued, to investigate the role of Lis1/dynactin in mitotic checkpoint function. In addition, we develop an improved time-lapse video microscopy technique that allows live imaging of GFP-Lis1, GFP-Rod checkpoint protein, green fluorescent protein (GFP)-labeled chromosomes, or GFP-labeled mitotic spindle dynamics in neuroblasts within whole larval brain explants. Our mutant analyses show that Lis1/dynactin have at least two independent functions during mitosis: first promoting centrosome separation and bipolar spindle assembly during prophase/prometaphase, and subsequently generating interkinetochore tension and transporting checkpoint proteins off kinetochores during metaphase, thus promoting timely anaphase onset. Furthermore, we show that Lis1/dynactin/dynein physically associate and colocalize on centrosomes, spindle MTs, and kinetochores, and that regulation of Lis1/dynactin kinetochore localization in Drosophila differs from both Caenorhabditis elegans and mammals. We conclude that Lis1/dynactin act together to regulate multiple, independent functions in mitotic cells, including spindle formation and cell cycle checkpoint release.

scholarly journals Embedding and Chemical Reactivation of Green Fluorescent Protein in the Whole Mouse Brain for Optical Micro-Imaging

2017 ◽  
Vol 11 ◽  
Author(s):  
Yadong Gang ◽  
Hongfu Zhou ◽  
Yao Jia ◽  
Ling Liu ◽  
Xiuli Liu ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Mouse Brain ◽  
Fluorescent Protein ◽  
Green Fluorescent

Visualizing the Effects of Chemicals On Hematopoietic Stem Cell Engraftment in an Endogenous Niche

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 506-506
Author(s):  
Owen J. Tamplin ◽  
Ellen Durand ◽  
Pulin Li ◽  
Leonard I. Zon
Keyword(s):  
Endothelial Cells ◽  
Stem Cell ◽  
Molecular Mechanisms ◽  
Transforming Growth Factor ◽  
Fluorescent Protein ◽  
Fetal Liver ◽  
Live Imaging ◽  
Hematopoietic Tissue ◽  
Hematopoietic Stem ◽  
Niche Formation

Abstract Abstract 506 Hematopoietic stem and progenitor cells (HSPC) self-renew and give rise to all blood cell types throughout adulthood. Definitive HSPC arise from the hemogenic endothelium of the dorsal aorta, are released into circulation, and then seed an intermediate hematopoietic tissue before colonizing the adult marrow. In mammals this intermediate tissue is the fetal liver, and in the zebrafish it is the caudal hematopoietic tissue (CHT), a vascular plexus in the ventral tail of the embryo. We have generated the first highly specific zebrafish transgenic reporter of HSPC, using the previously described mouse Runx1 +23 kb intronic enhancer driving GFP (or mCherry) fluorescent protein. We have demonstrated that these Runx1+23 positive cells are capable of long-term engraftment and multi-lineage contribution. Using time-lapse live imaging in the embryo, we followed HSPC as they migrate to the CHT niche. Together with endothelial (kdrl(flk1):DsRed2) or stromal (cxcl12a(sdf-1a):DsRed2) reporter lines, we could visualize stem cell behavior directly in the endogenous niche. Upon arrival, HSPC underwent a number of distinct steps to engraftment, including: 1) adherence to the vessel wall; 2) extravasation; 3) migration to the abluminal space; 4) triggering of niche formation—endothelial cells actually remodel around a single HSPC to create a niche; 5) cell division decisions. To determine if endothelial niche formation is conserved in mammals during ontogeny, we performed live imaging of mouse fetal liver explants at embryonic day 11.5, the earliest stage of seeding by HSPC. We observed rare c-kit+/Ly6a(Sca1):GFP+ HSPC become centered in a rosette of CD31+/Lyve1+ sinusoidal endothelial cells. This dynamic remodeling of endothelial cells around an HSPC in the niche was strikingly similar to the cellular behaviors we observed in zebrafish. We hypothesized that chemical genetics could reveal the molecular mechanisms and signaling pathways that are associated with the distinct steps of HSPC engraftment. As proof-of-concept, we tested the CXCR4 antagonist AMD3100 because the CXCR4-CXCL12 receptors and ligands are expressed in the CHT, and found that it suppressed CHT hematopoiesis. Next, we performed a chemical genetic screen by applying ∼2400 individual compounds of known action to zebrafish embryos during colonization of the CHT. We found 40 compounds that increased and 107 compounds that decreased CHT hematopoiesis. Applying selected chemical hits in our live imaging assay we found that certain compounds actually modulated distinct steps during engraftment. We identified a role for sphingosine-1-phosphate signaling during extravasation. We observed that regulators of the transcription factor hypoxia inducible factor (HIF)-1α modulated migration into abluminal spaces. The HIF-1α stabilizer dimethyloxalylglycine (DMOG) promoted migration into hypoxic abluminal spaces, while conversely the HIF-1α inhibitor YC-1 promoted migration into normoxic luminal spaces. We found the plant alkaloid Lycorine promoted endothelial niche formation, creating more locations for HSPC and allowing longer residence times in the CHT. The transforming growth factor (TGF)-β receptor inhibitor SB-431542 increased the rate of HSPC division after they had arrived in the niche. Our studies provide the first genetic approach to understanding engraftment, and the chemicals found could be used therapeutically for patients receiving marrow transplantation. Disclosures: Tamplin: Boston Children's Hospital: Employment, Patents & Royalties. Zon:Fate Therapeutics: Founder Other.

scholarly journals Live imaging using adaptive optics with fluorescent protein guide-stars

2012 ◽  
Vol 20 (14) ◽  
pp. 15969 ◽  
Author(s):  
Xiaodong Tao ◽  
Justin Crest ◽  
Shaila Kotadia ◽  
Oscar Azucena ◽  
Diana C. Chen ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Fluorescent Protein ◽  
Live Imaging
Sign in / Sign up

Export Citation Format

Share Document