scholarly journals Synchronized Formation and Remodeling of Postsynaptic Densities: Long-Term Visualization of Hippocampal Neurons Expressing Postsynaptic Density Proteins Tagged with Green Fluorescent Protein

2003 ◽  
Vol 23 (6) ◽  
pp. 2170-2181 ◽  
Author(s):  
Tatsuhiko Ebihara ◽  
Izumi Kawabata ◽  
Shinichi Usui ◽  
Kenji Sobue ◽  
Shigeo Okabe
Keyword(s):  
Green Fluorescent Protein ◽  
Hippocampal Neurons ◽  
Fluorescent Protein ◽  
Postsynaptic Density ◽  
Postsynaptic Density Proteins ◽  
Green Fluorescent

Functional expression of distinct NMDA channel subunits tagged with green fluorescent protein in hippocampal neurons in culture

2002 ◽  
Vol 42 (3) ◽  
pp. 306-318 ◽  
Author(s):  
J.-H. Luo ◽  
Z.Y. Fu ◽  
G. Losi ◽  
B.G. Kim ◽  
K. Prybylowski ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Hippocampal Neurons ◽  
Fluorescent Protein ◽  
Functional Expression ◽  
Nmda Channel ◽  
Green Fluorescent

scholarly journals Retrograde Transport of Transcription Factor NF-κB in Living Neurons

2000 ◽  
Vol 276 (15) ◽  
pp. 11821-11829 ◽  
Author(s):  
Henning Wellmann ◽  
Barbara Kaltschmidt ◽  
Christian Kaltschmidt
Keyword(s):  
Transcription Factor ◽  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Retrograde Transport ◽  
Receptor Activation ◽  
Transcriptional Changes ◽  
Localization Signal ◽  
Green Fluorescent ◽  
Living Neurons

The mechanism by which signals such as those produced by glutamate are transferred to the nucleus may involve direct transport of an activated transcription factor to trigger long-term transcriptional changes. Ionotropic glutamate receptor activation or depolarization activates transcription factor NF-κB and leads to translocation of NF-κB from the cytoplasm to the nucleus. We investigated the dynamics of NF-κB translocation in living neurons by tracing the NF-κB subunit RelA (p65) with jellyfish green fluorescent protein. We found that green fluorescent protein-RelA was located in either the nucleus or cytoplasm and neurites, depending on the coexpression of the cognate inhibitor of NF-κB, IκB-α. Stimulation with glutamate, kainate, or potassium chloride resulted in a redistribution of NF-κB from neurites to the nucleus. This transport depended on an intact nuclear localization signal on RelA. Thus, in addition to its role as a transcription factor, NF-κB may be a signal transducer, transmitting transient glutamatergic signals from distant sites to the nucleus.

scholarly journals Characterization of Leptin-Responsive Neurons in the Caudal Brainstem

Endocrinology ◽  
2006 ◽  
Vol 147 (7) ◽  
pp. 3190-3195 ◽  
Author(s):  
Kate L. J. Ellacott ◽  
Ilia G. Halatchev ◽  
Roger D. Cone
Keyword(s):  
Green Fluorescent Protein ◽  
Energy Homeostasis ◽  
Leptin Receptor ◽  
Fluorescent Protein ◽  
Physiological Role ◽  
Cell Group ◽  
Melanocortin System ◽  
Peptide Precursor ◽  
Green Fluorescent

The central melanocortin system plays a key role in the regulation of energy homeostasis. Neurons containing the peptide precursor proopiomelanocortin (POMC) are found at two sites in the brain, the arcuate nucleus of the hypothalamus (ARC) and the caudal region of the nucleus of the solitary tract (NTS). ARC POMC neurons, which also express cocaine- and amphetamine-regulated transcript (CART), are known to mediate part of the response to factors regulating energy homeostasis, such as leptin and ghrelin. In contrast, the physiological role(s) of the POMC neurons in the caudal brainstem are not well characterized. However, development of a transgenic mouse expressing green fluorescent protein under the control of the POMC promoter [POMC-enhanced green fluorescent protein (EGFP) mouse] has aided the study of these neurons. Indeed, recent studies have shown significant activation of NTS POMC-EGFP cells by the gut released satiety factor cholecystokinin (CCK). Here we show that peripheral leptin administration induces the expression of phospho-signal transducer and activator of transcription 3 immunoreactivity (pSTAT3-IR), a marker of leptin receptor signaling, in more than 50% of NTS POMC-EGFP neurons. Furthermore, these POMC-EGFP neurons comprise 30% of all pSTAT3-IR cells in the NTS. Additionally, we also show that in contrast to the ARC population, NTS POMC-EGFP neurons do not coexpress CART immunoreactivity. These data suggest that NTS POMC neurons may participate with ARC POMC cells in mediating some of the effects of leptin and thus comprise a novel cell group regulated by both long-term adipostatic signals and satiety factors such as CCK.

Analysis of GABAA Receptor Assembly in Mammalian Cell Lines and Hippocampal Neurons Using γ2 Subunit Green Fluorescent Protein Chimeras

2000 ◽  
Vol 16 (4) ◽  
pp. 440-452 ◽  
Author(s):  
Josef T. Kittler ◽  
Jianfeng Wang ◽  
Christopher N. Connolly ◽  
Stefano Vicini ◽  
Trevor G. Smart ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Gabaa Receptor ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Hippocampal Neurons ◽  
Fluorescent Protein ◽  
Mammalian Cell Lines ◽  
Green Fluorescent ◽  
Receptor Assembly

scholarly journals A neuron-specific cytoplasmic dynein isoform preferentially transports TrkB signaling endosomes

2008 ◽  
Vol 181 (6) ◽  
pp. 1027-1039 ◽  
Author(s):  
Junghoon Ha ◽  
Kevin W.-H. Lo ◽  
Kenneth R. Myers ◽  
Tiffany M. Carr ◽  
Michael K. Humsi ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Receptor Tyrosine Kinase ◽  
Pc12 Cell ◽  
Hippocampal Neurons ◽  
Fluorescent Protein ◽  
Cytoplasmic Dynein ◽  
Neurotrophin Receptor ◽  
Immunoaffinity Purification ◽  
Green Fluorescent ◽  
Cultured Hippocampal Neurons

Cytoplasmic dynein is the multisubunit motor protein for retrograde movement of diverse cargoes to microtubule minus ends. Here, we investigate the function of dynein variants, defined by different intermediate chain (IC) isoforms, by expressing fluorescent ICs in neuronal cells. Green fluorescent protein (GFP)–IC incorporates into functional dynein complexes that copurify with membranous organelles. In living PC12 cell neurites, GFP–dynein puncta travel in both the anterograde and retrograde directions. In cultured hippocampal neurons, neurotrophin receptor tyrosine kinase B (TrkB) signaling endosomes are transported by cytoplasmic dynein containing the neuron-specific IC-1B isoform and not by dynein containing the ubiquitous IC-2C isoform. Similarly, organelles containing TrkB isolated from brain by immunoaffinity purification also contain dynein with IC-1 but not IC-2 isoforms. These data demonstrate that the IC isoforms define dynein populations that are selectively recruited to transport distinct cargoes.

scholarly journals Introduction of Green Fluorescent Protein (GFP) into Hippocampal Neurons through Viral Infection

2010 ◽  
Vol 2010 (4) ◽  
pp. pdb.prot5406-pdb.prot5406 ◽  
Author(s):  
R. Malinow ◽  
Y. Hayashi ◽  
M. Maletic-Savatic ◽  
S. H. Zaman ◽  
J.-C. Poncer ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Viral Infection ◽  
Hippocampal Neurons ◽  
Fluorescent Protein ◽  
Green Fluorescent

High-Efficiency Transduction and Long-Term Gene Expression with a Murine Stem Cell Retroviral Vector Encoding the Green Fluorescent Protein in Human Marrow Stromal Cells

1999 ◽  
Vol 10 (7) ◽  
pp. 1163-1173 ◽  
Author(s):  
Jeffrey C. Marx ◽  
James A. Allay ◽  
Derek A. Persons ◽  
Sharon A. Nooner ◽  
Phillip W. Hargrove ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Green Fluorescent Protein ◽  
Stromal Cells ◽  
High Efficiency ◽  
Fluorescent Protein ◽  
Retroviral Vector ◽  
Marrow Stromal Cells ◽  
Green Fluorescent

scholarly journals Microtubule-dependent Recruitment of Staufen-Green Fluorescent Protein into Large RNA-containing Granules and Subsequent Dendritic Transport in Living Hippocampal Neurons

1999 ◽  
Vol 10 (9) ◽  
pp. 2945-2953 ◽  
Author(s):  
Martin Köhrmann ◽  
Ming Luo ◽  
Christoph Kaether ◽  
Luc DesGroseillers ◽  
Carlos G. Dotti ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Hippocampal Neurons ◽  
Rna Binding ◽  
Fluorescent Protein ◽  
Maximum Velocity ◽  
Time Lapse ◽  
Mrna Transport ◽  
Transport Pathway ◽  
Green Fluorescent ◽  
Dependent Transport

Dendritic mRNA transport and local translation at individual potentiated synapses may represent an elegant way to form synaptic memory. Recently, we characterized Staufen, a double-stranded RNA-binding protein, in rat hippocampal neurons and showed its presence in large RNA-containing granules, which colocalize with microtubules in dendrites. In this paper, we transiently transfect hippocampal neurons with human Staufen-green fluorescent protein (GFP) and find fluorescent granules in the somatodendritic domain of these cells. Human Stau-GFP granules show the same cellular distribution and size and also contain RNA, as already shown for the endogenous Stau particles. In time-lapse videomicroscopy, we show the bidirectional movement of these Staufen-GFP–labeled granules from the cell body into dendrites and vice versa. The average speed of these particles was 6.4 μm/min with a maximum velocity of 24.3 μm/min. Moreover, we demonstrate that the observed assembly into granules and their subsequent dendritic movement is microtubule dependent. Taken together, we have characterized a novel, nonvesicular, microtubule-dependent transport pathway involving RNA-containing granules with Staufen as a core component. This is the first demonstration in living neurons of movement of an essential protein constituent of the mRNA transport machinery.

LONG-TERM EXPRESSION OF THE GENE ENCODING GREEN FLUORESCENT PROTEIN IN MURINE HEMATOPOIETIC CELLS USING RETROVIRAL GENE TRANSFER1

1998 ◽  
Vol 65 (9) ◽  
pp. 1233-1240 ◽  
Author(s):  
Jessamyn Bagley ◽  
Karen Aboody-Guterman ◽  
Xandra Breakefield ◽  
John Iacomini
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Hematopoietic Cells ◽  
Gene Encoding ◽  
Green Fluorescent
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