scholarly journals Transgenic Mice Expressing Green Fluorescent Protein under the Control of the Corticotropin-Releasing Hormone Promoter

Endocrinology ◽  
2009 ◽  
Vol 150 (12) ◽  
pp. 5626-5632 ◽  
Author(s):  
Tamar Alon ◽  
Ligang Zhou ◽  
Cristian A. Pérez ◽  
Alastair S. Garfield ◽  
Jeffrey M. Friedman ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Artificial Chromosome ◽  
Brain Regions ◽  
Ingestive Behavior ◽  
Physiological Processes ◽  
Functional Relevance ◽  
Green Fluorescent ◽  
And Function ◽  
The Brain

Abstract CRH is widely expressed in the brain and is of broad functional relevance to a number of physiological processes, including stress response, parturition, immune response, and ingestive behavior. To delineate further the organization of the central CRH network, we generated mice expressing green fluorescent protein (GFP) under the control of the CRH promoter, using bacterial artificial chromosome technology. Here we validate CRH-GFP transgene expression within specific brain regions and confirm the distribution of central GFP-producing cells to faithfully recapitulate that of CRH-expressing cells. Furthermore, we confirm the functional integrity of a population of GFP-producing cells by demonstrating their apposite responsiveness to nutritional status. We anticipate that this transgenic model will lend itself as a highly tractable tool for the investigation of CRH expression and function in discrete brain regions.

Serca2a-Phospholamban Interaction Monitored by an Interposed Circularly Permutated Green Fluorescent Protein

2021 ◽  
Author(s):  
Maren E Arnold ◽  
Wolfgang R Dostmann ◽  
Jody Martin ◽  
Michael J Previs ◽  
Bradley Palmer ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Recombinant Protein ◽  
Integrated Assessment ◽  
Fluorescent Protein ◽  
Cardiac Contractility ◽  
Fluorescence Yield ◽  
Selective Interference ◽  
General Activation ◽  
Green Fluorescent ◽  
And Function

Background: The interaction of phospholamban (PLB) and the sarcoplasmic reticulum Ca2+-ATPase (Serca2a) is a key regulator of cardiac contractility and a therapeutic target in heart failure (HF). PLB mediated increases in Serca2a activity improve cardiac function and HF. Clinically this mechanism can only be exploited by a general activation of the proteinkinase A (PKA) which is associated with side effects and adverse clinical outcomes. A selective interference of the PLB-Serca2a interaction is desirable but will require novel tools that allow for an integrated assessment of this interaction under both physiological and pathophysiological conditions. Methods: A circularly permutated green fluorescent protein (cpGFP) was interposed between Serca2a and PLB to result into a single Serca2a-cpGFP-PLB recombinant protein (SGP). Expression, phosphorylation, fluorescence and function of SGP were evaluated. Results: Expression of SGP-cDNA results in a functional recombinant protein at the predicted molecular weight. The PLB domain of SGP retains its ability to polymerize and can be phosphorylated by PKA activation. This increases the fluorescent yield of SGP by between 10% to 165% depending on cell line and conditions. Summary: A single recombinant fusion protein that combines Serca2a, a circularly permutated green fluorescent protein and PLB can be expressed in cells and can be phosphorylated at the PLB domain which markedly increases the fluorescence yield. SGP is a novel cellular Serca2a-PLB interaction monitor.

scholarly journals Visualization of lymphatic vessels by Prox1-promoter directed GFP reporter in a bacterial artificial chromosome-based transgenic mouse

Blood ◽  
2011 ◽  
Vol 117 (1) ◽  
pp. 362-365 ◽  
Author(s):  
Inho Choi ◽  
Hee Kyoung Chung ◽  
Swapnika Ramu ◽  
Ha Neul Lee ◽  
Kyu Eui Kim ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Bacterial Artificial Chromosome ◽  
Transgenic Mouse ◽  
Fluorescent Protein ◽  
Lymphatic Vessels ◽  
Artificial Chromosome ◽  
Transgenic Animal ◽  
Lymphatic Development ◽  
Vasculogenesis And Angiogenesis ◽  
Green Fluorescent

Abstract Although the blood vessel-specific fluorescent transgenic mouse has been an excellent tool to study vasculogenesis and angiogenesis, a lymphatic-specific fluorescent mouse model has not been established to date. Here we report a transgenic animal model that expresses the green fluorescent protein under the promoter of Prox1, a master control gene in lymphatic development. Generated using an approximately 200-kb-long bacterial artificial chromosome harboring the entire Prox1 gene, this Prox1-green fluorescent protein mouse was found to faithfully recapitulate the expression pattern of the Prox1 gene in lymphatic endothelial cells and other Prox1-expressing organs, and enabled us to conveniently visualize detailed structure and morphology of lymphatic vessels and networks throughout development. Our data demonstrate that this novel transgenic mouse can be extremely useful for detection, imaging, and isolation of lymphatic vessels and monitoring wound-associated lymphangiogenesis. Together, this Prox1-green fluorescent protein transgenic mouse will be a great tool for the lymphatic research.

scholarly journals TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2751
Author(s):  
Osung Kwon ◽  
Hayoung Yang ◽  
Seung-Chan Kim ◽  
Juhyun Kim ◽  
Jaewon Sim ◽  
...  
Keyword(s):  
Animal Model ◽  
Neuronal Excitability ◽  
Fluorescent Protein ◽  
Granule Cells ◽  
Expression Patterns ◽  
Artificial Chromosome ◽  
Lack Of Information ◽  
Green Fluorescent ◽  
Pore Domain ◽  
The Brain

TWIK-1 is the first identified member of the two-pore domain potassium (K2P) channels that are involved in neuronal excitability and astrocytic passive conductance in the brain. Despite the physiological roles of TWIK-1, there is still a lack of information on the basic expression patterns of TWIK-1 proteins in the brain. Here, using a modified bacterial artificial chromosome (BAC), we generated a transgenic mouse (Tg mouse) line expressing green fluorescent protein (GFP) under the control of the TWIK-1 promoter (TWIK-1 BAC-GFP Tg mice). We confirmed that nearly all GFP-producing cells co-expressed endogenous TWIK-1 in the brain of TWIK-1 BAC-GFP Tg mice. GFP signals were highly expressed in various brain areas, including the dentate gyrus (DG), lateral entorhinal cortex (LEC), and cerebellum (Cb). In addition, we found that GFP signals were highly expressed in immature granule cells in the DG. Finally, our TWIK-1 BAC-GFP Tg mice mimic the upregulation of TWIK-1 mRNA expression in the hippocampus following the injection of kainic acid (KA). Our data clearly showed that TWIK-1 BAC-GFP Tg mice are a useful animal model for studying the mechanisms regulating TWIK-1 gene expression and the physiological roles of TWIK-1 channels in the brain.

scholarly journals Live imaging of avian embryos revealing a new head precursor map and the role for the anterior mesendoderm in brain development

2020 ◽  
Author(s):  
Koya Yoshihi ◽  
Kagayaki Kato ◽  
Hideaki Iida ◽  
Machiko Teramoto ◽  
Akihito Kawamura ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Brain Development ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Cellular Mechanisms ◽  
Avian Embryos ◽  
Head Development ◽  
Green Fluorescent ◽  
The Brain ◽  
Graft Positioning

ABSTRACTWe investigated the initial stages of head development using a new method to randomly label chicken epiblast cells with enhanced green fluorescent protein, and tracking the labeled cells. This analysis was combined with grafting mCherry-expressing quail nodes, or node-derived anterior mesendoderm (AME). These live imagings provided a new conception of the cellular mechanisms regulating brain and head ectoderm development. Virtually all anterior epiblast cells are bipotent for the development into the brain or head ectoderm. Their fate depends on the positioning after converging to the AME. When two AME tissues exist following the ectopic node graft, the epiblast cells converge to the two AME positions and develop into two brain tissues. The anterior epiblast cells bear gross regionalities that already correspond to the forebrain, midbrain, and hindbrain axial levels shortly after the node is formed. Therefore, brain portions that develop with the graft-derived AME are dependent on graft positioning.

Visualization of glucocorticoid receptor in the brain of green fluorescent protein–glucocorticoid receptor knockin mice

Neuroscience ◽  
2005 ◽  
Vol 135 (4) ◽  
pp. 1119-1128 ◽  
Author(s):  
T. Usuku ◽  
M. Nishi ◽  
M. Morimoto ◽  
J.A. Brewer ◽  
L.J. Muglia ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Glucocorticoid Receptor ◽  
Fluorescent Protein ◽  
Green Fluorescent ◽  
The Brain

scholarly journals Construction and function of a recombinant adenovirus encoding a human aquaporin 1-green fluorescent protein fusion product

2000 ◽  
Vol 7 (3) ◽  
pp. 476-485 ◽  
Author(s):  
A T M Shamsul Hoque ◽  
Xibao Liu ◽  
Hideaki Kagami ◽  
William D Swaim ◽  
Robert B Wellner ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Recombinant Adenovirus ◽  
Fusion Product ◽  
Green Fluorescent Protein Fusion ◽  
Protein Fusion ◽  
Aquaporin 1 ◽  
Green Fluorescent ◽  
And Function

scholarly journals Survival and Immunogenicity of Mesenchymal Stem Cells From the Green Fluorescent Protein Transgenic Rat in the Adult Rat Brain

2010 ◽  
Vol 24 (7) ◽  
pp. 645-656 ◽  
Author(s):  
Teresa C. Moloney ◽  
Peter Dockery ◽  
Anthony J. Windebank ◽  
Frank P. Barry ◽  
Linda Howard ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Adult Brain ◽  
Transgenic Rat ◽  
Reporter Protein ◽  
Technical Limitation ◽  
Green Fluorescent ◽  
The Brain

Background. A major technical limitation in preclinical cell replacement research is the ability to discriminate between donor and host tissue after transplantation. This problem has been lessened by the availability of transgenic animals that express “reporter” genes, such as green fluorescent protein (GFP). Objective. We determined the usefulness of one such transgenic reporter rat to assess the survival of bone marrow—derived rat mesenchymal stem cells (MSCs) following direct transplantation into the intact adult brain. We also sought to determine if the expression of GFP in the brain affected the survival of the MSCs or the host’s neuroimmune response to the cells. Methods. Rats received intrastriatal injections of sterile transplantation medium, 100 000 normal MSCs, or 100 000 GFP-MSCs and were killed humanely 1, 4, 7, 28, and 42 days posttransplantation for astrocyte and microglial immunohistochemical staining. Results. GFP-MSCs were evident at each examination, although their survival declined over time. Graft volume estimates comparing normal and GFP-MSCs revealed that GFP expression did not adversely affect the survival of the stem cells in the brain. Furthermore, immunostaining for astrocytes and microglia revealed that expression of the reporter protein did not affect the immunogenicity of the stem cells. Conclusions. These data indicate the usefulness of GFP for investigating the survival of MSCs following transplantation to the brain. However, the mechanisms responsible for the poor survival of the stem cells must be elucidated if these cells are to serve cell-based therapies for neurodegenerative disorders.

Measurements of vector-derived neurotrophic factor and green fluorescent protein levels in the brain

Methods ◽  
2002 ◽  
Vol 28 (2) ◽  
pp. 286-292 ◽  
Author(s):  
Ronald L. Klein ◽  
Mary E. Hamby ◽  
Christopher F. Sonntag ◽  
William J. Millard ◽  
Michael A. King ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Neurotrophic Factor ◽  
Fluorescent Protein ◽  
Protein Levels ◽  
Green Fluorescent ◽  
The Brain

scholarly journals The jellyfish green fluorescent protein: A new tool for studying ion channel expression and function

Neuron ◽  
1995 ◽  
Vol 14 (2) ◽  
pp. 211-215 ◽  
Author(s):  
John Marshall ◽  
Raymond Molloy ◽  
Guy W.J Moss ◽  
James R Howe ◽  
Thomas E Hughes
Keyword(s):  
Green Fluorescent Protein ◽  
Ion Channel ◽  
Fluorescent Protein ◽  
Protein A ◽  
Channel Expression ◽  
Green Fluorescent ◽  
And Function

scholarly journals Astrocyte-to-neuron transportation of Enhanced Green Fluorescent Protein in cerebral cortex requires F-actin dependent Tunneling Nanotubes

2021 ◽  
Author(s):  
Jing Chen ◽  
Junyan Cao
Keyword(s):  
Cerebral Cortex ◽  
Green Fluorescent Protein ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Cell Contact ◽  
Neural Cells ◽  
Tunneling Nanotubes ◽  
Green Fluorescent ◽  
And Function

Abstract Tunneling Nanotube (TNT) , a dynamic cell-cell contact, is dependent on actin polimerization. TNTs are efficient in transporting ions, proteins and organelles intercellularly, which are important mechanisms in physiological and phathological precesses. Reported studies on the existence and function of TNTs among neural cells focus on cultured cell for the convenience in detecting TNTs’ ultrastructure. In this study, the adeno-associated virus (AAV-GFAP-EGFP-p2A-cre) was injected in the cerebral cortex of knock-in mice ROSA26 GNZ. GFAP promoter initiated the expression of Enhanced Green Fluorescent Protein (EGFP) in infected astrocytes. At 10 days post injection (10 DPI), we found that EGFP could transfer from astrocytes in layerⅠ-Ⅲ to neurons in layer Ⅴ. The dissemination of EGFP was not through endocytosis or exosome. And the intercellular transportation of EGFP was F-actin dependent. Therefore, we concluded EGFP transported from astrocytes to neurons in coetex via F-actin dependent TNTs. Although it is hardly to detect the ultrastrucute of TNTs in brain for its transiency and the noisy background, we established an animal model and indirect experimental methods to explore TNTs in vivo.

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