| Robust up-regulation of nuclear red fluorescent-tagged fos marks neuronal activation in green fluorescent vasopressin neurons after osmotic stimulation in a double-transgenic rat. | |
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MedLine Citation:
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PMID: 19850746 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The up-regulation in the expression of mRNA or protein encoded by the c-fos gene is widely used as a marker of neuronal activation elicited by various stimuli. To facilitate the detection of activated neurons, we generated transgenic rats expressing a fusion gene consisting of c-fos coding sequences in frame with monomeric red fluorescent protein 1 (mRFP1) under the control of c-fos gene regulatory sequences (c-fos-mRFP1 rats). In c-fos-mRFP1 transgenic rats, 90 min after hypertonic saline ip administration, nuclear mRFP1 fluorescence was observed abundantly in brain regions known to be osmosensitive, namely the median preoptic nucleus, organum vasculosum lamina terminalis, supraoptic nucleus, paraventricular nucleus, and subfornical organ. Immunohistochemistry for Fos protein confirmed that the distribution of Fos-like immunoreactivity in nontransgenic rats was similar to those of mRFP1 fluorescence after ip administration of hypertonic saline in the transgenic rats. Several double-transgenic rats were obtained from matings between transgenic rats expressing an arginine vasopressin-enhanced green fluorescent protein fusion gene (AVP-eGFP rats) and c-fos-mRFP1 rats. In these double-transgenic rats, almost all eGFP neurons in the supraoptic nucleus and PVN expressed nuclear mRFP1 fluorescence 90 min after hypertonic saline administration. The c-fos-mRFP1 rats are a powerful tool that enables the facile identification of activated neurons in the nervous system. Furthermore, when combined with transgenes expressing another fluorophore under the control of cell-specific regulatory sequences, activation of specific neuronal cell types in response to physiological cues can be readily detected. |
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Authors:
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Hiroaki Fujihara; Yoichi Ueta; Hitoshi Suzuki; Akiko Katoh; Toyoaki Ohbuchi; Hiroki Otsubo; Govindan Dayanithi; David Murphy |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-10-22 |
Journal Detail:
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Title: Endocrinology Volume: 150 ISSN: 1945-7170 ISO Abbreviation: Endocrinology Publication Date: 2009 Dec |
Date Detail:
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Created Date: 2009-11-25 Completed Date: 2010-01-19 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0375040 Medline TA: Endocrinology Country: United States |
Other Details:
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Languages: eng Pagination: 5633-8 Citation Subset: AIM; IM |
Affiliation:
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Department of Physiology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Arginine Vasopressin / genetics, metabolism* Brain / cytology, metabolism* Female Green Fluorescent Proteins / genetics, metabolism Hypothalamus / cytology, metabolism Immunohistochemistry Luminescent Proteins / genetics, metabolism Male Mice Mice, Inbred C57BL Microscopy, Confocal Microscopy, Fluorescence Neurons / metabolism* Paraventricular Hypothalamic Nucleus / cytology, metabolism Proto-Oncogene Proteins c-fos / genetics, metabolism* Rats Rats, Transgenic Rats, Wistar Recombinant Fusion Proteins / genetics, metabolism Regulatory Sequences, Nucleic Acid / genetics Saline Solution, Hypertonic / administration & dosage, pharmacology Subfornical Organ / cytology, metabolism Supraoptic Nucleus / cytology, metabolism Up-Regulation / drug effects |
| Chemical | |
Reg. No./Substance:
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0/Luminescent Proteins; 0/Proto-Oncogene Proteins c-fos; 0/Recombinant Fusion Proteins; 0/Saline Solution, Hypertonic; 0/red fluorescent protein; 113-79-1/Arginine Vasopressin; 147336-22-9/Green Fluorescent Proteins |
| Comments/Corrections | |
Comment In:
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Endocrinology. 2009 Dec;150(12):5199-201
[PMID:
19933396
]
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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