Analysis of Cell Movement Between Skin and Other Anatomical Sites In Vivo Using Photoconvertible Fluorescent Protein “Kaede”-Transgenic Mice

2012 ◽  
pp. 279-286 ◽  
Author(s):  
Michio Tomura ◽  
Kenji Kabashima
Keyword(s):  
Transgenic Mice ◽  
Fluorescent Protein ◽  
Cell Movement

scholarly journals Fluorescently Tagged Potato virus Y: A Versatile Tool for Functional Analysis of Plant-Virus Interactions

2015 ◽  
Vol 28 (7) ◽  
pp. 739-750 ◽  
Author(s):  
Matevz Rupar ◽  
Florence Faurez ◽  
Michel Tribodet ◽  
Ion Gutiérrez-Aguirre ◽  
Agnès Delaunay ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Potato Virus ◽  
Plant Virus ◽  
Potato Virus Y ◽  
Fluorescent Protein ◽  
Ex Vivo ◽  
Cell Movement ◽  
Long Distance ◽  
Green Fluorescent

Potato virus Y (PVY) is an economically important plant virus that infects Solanaceous crops such as tobacco and potato. To date, studies into the localization and movement of PVY in plants have been limited to detection of viral RNA or proteins ex vivo. Here, a PVY N605 isolate was tagged with green fluorescent protein (GFP), characterized and used for in vivo tracking. In Nicotiana tabacum cv. Xanthi, PVY N605-GFP was biologically comparable to nontagged PVY N605, stable through three plant-to-plant passages and persisted for four months in infected plants. GFP was detected before symptoms and fluorescence intensity correlated with PVY RNA concentrations. PVY N605-GFP provided in vivo tracking of long-distance movement, allowing estimation of the cell-to-cell movement rate of PVY in N. tabacum cv. Xanthi (7.1 ± 1.5 cells per hour). PVY N605-GFP was adequately stable in Solanum tuberosum cvs. Désirée and NahG-Désirée and able to infect S. tuberosum cvs. Bintje and Bea, Nicotiana benthamiana, and wild potato relatives. PVY N605-GFP is therefore a powerful tool for future studies of PVY-host interactions, such as functional analysis of viral and plant genes involved in viral movement.

scholarly journals Neuroendocrine Plasticity in the Anterior Pituitary: Gonadotropin-Releasing Hormone-Mediated Movement in Vitro and in Vivo

Endocrinology ◽  
2007 ◽  
Vol 148 (4) ◽  
pp. 1736-1744 ◽  
Author(s):  
Amy M. Navratil ◽  
J. Gabriel Knoll ◽  
Jennifer D. Whitesell ◽  
Stuart A. Tobet ◽  
Colin M. Clay
Keyword(s):  
Pituitary Gland ◽  
Anterior Pituitary ◽  
Fluorescent Protein ◽  
Cell Movement ◽  
Pituitary Cells ◽  
Rous Sarcoma ◽  
Cytoskeleton Disruption ◽  
Green Fluorescent

The secretion of LH is cued by the hypothalamic neuropeptide, GnRH. After delivery to the anterior pituitary gland via the hypothalamic-pituitary portal vasculature, GnRH binds to specific high-affinity receptors on the surface of gonadotrope cells and stimulates synthesis and secretion of the gonadotropins, FSH, and LH. In the current study, GnRH caused acute and dramatic changes in cellular morphology in the gonadotrope-derived αT3-1 cell line, which appeared to be mediated by engagement of the actin cytoskeleton; disruption of actin with jasplakinolide abrogated cell movement and GnRH-induced activation of ERK. In live murine pituitary slices infected with an adenovirus-containing Rous sarcoma virus-green fluorescent protein, selected cells responded to GnRH by altering their cellular movements characterized by both formation and extension of cell processes and, surprisingly, spatial repositioning. Consistent with the latter observation, GnRH stimulation increased the migration of dissociated pituitary cells in transwell chambers. Our data using live pituitary slices are a striking example of neuropeptide-evoked movements of cells outside the central nervous system and in a mature peripheral endocrine organ. These findings call for a fundamental change in the current dogma of simple passive diffusion of LH from gonadotropes to capillaries in the pituitary gland.

scholarly journals GFP transgenic animals in biomedical research: a review of potential disadvantages

2019 ◽  
pp. 525-530
Author(s):  
N. Lipták ◽  
Z. Bősze ◽  
L. Hiripi
Keyword(s):  
Transgenic Mice ◽  
Biomedical Research ◽  
Fluorescent Protein ◽  
Yellow Fluorescent Protein ◽  
Transgenic Animals ◽  
In Vivo Studies ◽  
Reporter Protein ◽  
Physiological Processes ◽  
Green Fluorescent

Green Fluorescent protein (GFP) transgenic animals are accepted tools for studying various physiological processes, including organ development and cell migration. However, several in vivo studies claimed that GFP may impair transgenic animals’ health. Glomerulosclerosis was observed in transgenic mice and rabbits with ubiquitous reporter protein expression. Heart-specific GFP expression evoked dilated cardiomyopathy and altered cardiac function in transgenic mouse and zebrafish lines, respectively. Moreover, growth retardation and increased axon swelling were observed in GFP and yellow fluorescent protein (YFP) transgenic mice, respectively. This review will focus on the potential drawbacks of the applications of GFP transgenic animals in biomedical research.

scholarly journals A Novel Platform to Test In Vivo Single Gene Dependencies in t(8,21) and t(15,17) AML Confirms Zeb2 as Leukemia Target

Cancers ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3768
Author(s):  
Giulia De Conti ◽  
Alicja M. Gruszka ◽  
Debora Valli ◽  
Andrea Umberto Cammarata ◽  
Matteo Righi ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Fluorescent Protein ◽  
Constitutive Expression ◽  
Model Systems ◽  
Leukemic Cells ◽  
Tamoxifen Treatment ◽  
Selection Marker ◽  
In Vivo Model ◽  
Hematopoietic Stem

The increased usage of high-throughput technologies in cancer research, including genetic and drug screens, generates large sets of candidate targets that need to be functionally validated for their roles in tumor development. Thus, reliable and robust in vivo model systems are needed to perform reverse genetic experiments. Ideally, these models should allow for a conditional silencing of the target and an unambiguous identification of engineered cancer cells. Here, we present a platform consisting of: (i) t(8;21) and t(15;17) driven acute myeloid leukemia (AML) transgenic mice with constitutive expression of green fluorescent protein (GFP) and inducible expression of Cre recombinase, and (ii) REX, a modified pSico lentiviral vector for inducible shRNA expression and red fluorescent protein (RFP) as a selection marker. In this system, leukemic cells from transgenic mice are transduced with REX, flow sorted, and transplanted into syngeneic hosts. Gene interference is induced in established tumors by tamoxifen treatment. Dual-color cell fluorescence guides the in vivo identification of shRNA interfered AML cells, monitoring engraftment and disease progression. We tested the platform by inducing knockdown of Zeb2, a gene upregulated by AML1-ETO and PML-RARα oncogenes in pre-leukemic hematopoietic stem cell compartment, and observed a significant delay in leukemia onset. This proves the power and utility of the platform and confirms Zeb2 contribution to the pathogenesis of AML.

scholarly journals Downregulation of the NbNACa1 Gene Encoding a Movement-Protein-Interacting Protein Reduces Cell-to-Cell Movement of Brome mosaic virus in Nicotiana benthamiana

2007 ◽  
Vol 20 (6) ◽  
pp. 671-681 ◽  
Author(s):  
Masanori Kaido ◽  
Yosuke Inoue ◽  
Yoshika Takeda ◽  
Kazuhiko Sugiyama ◽  
Atsushi Takeda ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Movement Protein ◽  
Sequence Similarity ◽  
Microprojectile Bombardment ◽  
Cell Movement ◽  
Brome Mosaic Virus ◽  
Viral Movement

The 3a movement protein (MP) plays a central role in the movement of the RNA plant virus, Brome mosaic virus (BMV). To identify host factor genes involved in viral movement, a cDNA library of Nicotiana benthamiana, a systemic host for BMV, was screened with far-Western blotting using a recombinant BMV MP as probe. One positive clone encoded a protein with sequence similarity to the α chain of nascent-polypeptide-associated complex from various organisms, which is proposed to contribute to the fidelity of translocation of newly synthesized proteins. The orthologous gene from N. benthamiana was designated NbNACa1. The binding of NbNACa1 to BMV MP was confirmed in vivo with an agroinfiltration-immunoprecipitation assay. To investigate the involvement of NbNACa1 in BMV multiplication, NbNACa1-silenced (GSNAC) transgenic N. benthamiana plants were produced. Downregulation of NbNACa1 expression reduced virus accumulation in inoculated leaves but not in protoplasts. A microprojectile bombardment assay to monitor BMV-MP-assisted viral movement demonstrated reduced virus spread in GSNAC plants. The localization to the cell wall of BMV MP fused to green fluorescent protein was delayed in GSNAC plants. From these results, we propose that NbNACa1 is involved in BMV cell-to-cell movement through the regulation of BMV MP localization to the plasmodesmata.

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