000007022 001__ 7022
000007022 005__ 20180208231257.0
000007022 0247_ $$2pmid$$apmid:19761793
000007022 0247_ $$2DOI$$a10.1016/j.jneumeth.2009.09.010
000007022 0247_ $$2WOS$$aWOS:000272473000008
000007022 037__ $$aPreJuSER-7022
000007022 041__ $$aeng
000007022 082__ $$a610
000007022 084__ $$2WoS$$aBiochemical Research Methods
000007022 084__ $$2WoS$$aNeurosciences
000007022 1001_ $$0P:(DE-Juel1)VDB71164$$aCremer, C.M.$$b0$$uFZJ
000007022 245__ $$aFast, quantitative in situ hybridization of rare mRNAs using 14C-standards and phosphorus imaging
000007022 260__ $$aAmsterdam [u.a.]$$bElsevier Science$$c2009
000007022 300__ $$a56 - 61
000007022 3367_ $$0PUB:(DE-HGF)16$$2PUB:(DE-HGF)$$aJournal Article
000007022 3367_ $$2DataCite$$aOutput Types/Journal article
000007022 3367_ $$00$$2EndNote$$aJournal Article
000007022 3367_ $$2BibTeX$$aARTICLE
000007022 3367_ $$2ORCID$$aJOURNAL_ARTICLE
000007022 3367_ $$2DRIVER$$aarticle
000007022 440_0 $$09911$$aJournal of Neuroscience Methods$$v185$$x0165-0270$$y1
000007022 500__ $$aRecord converted from VDB: 12.11.2012
000007022 520__ $$aThe use of radiolabelled probes for in situ hybridization (ISH) bears the advantage of high sensitivity and quantifiability. The crucial disadvantages are laborious hybridization protocols, exposition of hybridized sections to film for up to several weeks and the time consuming need to prepare tissue standards with relatively short-lived isotopes like (33)P or (35)S for each experiment. The quantification of rare mRNAs like those encoding for subunits of neurotransmitter receptors is therefore a challenge in ISH. Here, we describe a method for fast, quantitative in situ hybridization (qISH) of mRNAs using (33)P-labelled oligonucleotides together with (14)C-polymer standards (Microscales, Amersham Biosciences) and a phosphorus imaging system (BAS 5000 BioImage Analyzer, Raytest-Fuji). It enables a complete analysis of rare mRNAs by ISH. The preparation of short-lived (33)P-standards for each experiment was replaced by co-exposition and calibration of long-lived (14)C-standards together with (33)P-labelled brain paste standards. The use of a phosphorus imaging system allowed a reduction of exposition time following hybridization from several weeks to a few hours or days. We used this approach as an example for applications to quantify the expression of GluR1 and GluR2 subunit mRNAs of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor in the hippocampus of untreated rats, and after intraperitoneal application of the organo-arsenic compound dimethyl arsenic acid.
000007022 536__ $$0G:(DE-Juel1)FUEK409$$2G:(DE-HGF)$$aFunktion und Dysfunktion des Nervensystems$$cP33$$x0
000007022 588__ $$aDataset connected to Web of Science, Pubmed
000007022 650_2 $$2MeSH$$aAnimals
000007022 650_2 $$2MeSH$$aCacodylic Acid: toxicity
000007022 650_2 $$2MeSH$$aCarbon Radioisotopes: chemistry
000007022 650_2 $$2MeSH$$aHippocampus: drug effects
000007022 650_2 $$2MeSH$$aHippocampus: metabolism
000007022 650_2 $$2MeSH$$aHippocampus: physiopathology
000007022 650_2 $$2MeSH$$aImage Processing, Computer-Assisted: instrumentation
000007022 650_2 $$2MeSH$$aImage Processing, Computer-Assisted: methods
000007022 650_2 $$2MeSH$$aIn Situ Hybridization: methods
000007022 650_2 $$2MeSH$$aMale
000007022 650_2 $$2MeSH$$aNeurochemistry: methods
000007022 650_2 $$2MeSH$$aOligonucleotides: chemistry
000007022 650_2 $$2MeSH$$aPhosphorus Radioisotopes: chemistry
000007022 650_2 $$2MeSH$$aRNA, Messenger: analysis
000007022 650_2 $$2MeSH$$aRNA, Messenger: metabolism
000007022 650_2 $$2MeSH$$aRats
000007022 650_2 $$2MeSH$$aRats, Wistar
000007022 650_2 $$2MeSH$$aReceptors, AMPA: genetics
000007022 650_2 $$2MeSH$$aReference Standards
000007022 650_2 $$2MeSH$$aTime Factors
000007022 650_7 $$00$$2NLM Chemicals$$aCarbon Radioisotopes
000007022 650_7 $$00$$2NLM Chemicals$$aOligonucleotides
000007022 650_7 $$00$$2NLM Chemicals$$aPhosphorus Radioisotopes
000007022 650_7 $$00$$2NLM Chemicals$$aRNA, Messenger
000007022 650_7 $$00$$2NLM Chemicals$$aReceptors, AMPA
000007022 650_7 $$00$$2NLM Chemicals$$aglutamate receptor ionotropic, AMPA 1
000007022 650_7 $$00$$2NLM Chemicals$$aglutamate receptor ionotropic, AMPA 2
000007022 650_7 $$075-60-5$$2NLM Chemicals$$aCacodylic Acid
000007022 650_7 $$2WoSType$$aJ
000007022 65320 $$2Author$$aQuantitative in situ hybridization
000007022 65320 $$2Author$$aStandardization
000007022 65320 $$2Author$$aMicroscales
000007022 65320 $$2Author$$aPhosphorus imaging
000007022 65320 $$2Author$$aAutoradiography
000007022 65320 $$2Author$$aNeurotransmitter receptors
000007022 65320 $$2Author$$aDimethyl arsenic acid
000007022 7001_ $$0P:(DE-Juel1)VDB21539$$aCremer, M.$$b1$$uFZJ
000007022 7001_ $$0P:(DE-Juel1)VDB89117$$aLopez Escobar, J.$$b2$$uFZJ
000007022 7001_ $$0P:(DE-HGF)0$$aSpeckmann, E.J.$$b3
000007022 7001_ $$0P:(DE-Juel1)131714$$aZilles, K.$$b4$$uFZJ
000007022 773__ $$0PERI:(DE-600)1500499-5$$a10.1016/j.jneumeth.2009.09.010$$gVol. 185, p. 56 - 61$$p56 - 61$$q185<56 - 61$$tJournal of neuroscience methods$$v185$$x0165-0270$$y2009
000007022 8567_ $$uhttp://dx.doi.org/10.1016/j.jneumeth.2009.09.010
000007022 909CO $$ooai:juser.fz-juelich.de:7022$$pVDB
000007022 9131_ $$0G:(DE-Juel1)FUEK409$$bGesundheit$$kP33$$lFunktion und Dysfunktion des Nervensystems$$vFunktion und Dysfunktion des Nervensystems$$x0
000007022 9141_ $$y2009
000007022 915__ $$0StatID:(DE-HGF)0010$$aJCR/ISI refereed
000007022 9201_ $$0I:(DE-Juel1)INM-2-20090406$$gINM$$kINM-2$$lMolekulare Organisation des Gehirns$$x0
000007022 9201_ $$0I:(DE-82)080010_20140620$$gJARA$$kJARA-BRAIN$$lJülich-Aachen Research Alliance - Translational Brain Medicine$$x1
000007022 970__ $$aVDB:(DE-Juel1)115510
000007022 980__ $$aVDB
000007022 980__ $$aConvertedRecord
000007022 980__ $$ajournal
000007022 980__ $$aI:(DE-Juel1)INM-2-20090406
000007022 980__ $$aI:(DE-82)080010_20140620
000007022 980__ $$aUNRESTRICTED
000007022 981__ $$aI:(DE-Juel1)VDB1046