Hauptseite > Publikationsdatenbank > Fast, quantitative in situ hybridization of rare mRNAs using 14C-standards and phosphorus imaging > print

001     7022
005     20180208231257.0
024 7 _ |2 pmid
|a pmid:19761793
024 7 _ |2 DOI
|a 10.1016/j.jneumeth.2009.09.010
024 7 _ |2 WOS
|a WOS:000272473000008
037 _ _ |a PreJuSER-7022
041 _ _ |a eng
082 _ _ |a 610
084 _ _ |2 WoS
|a Biochemical Research Methods
084 _ _ |2 WoS
|a Neurosciences
100 1 _ |a Cremer, C.M.
|b 0
|u FZJ
|0 P:(DE-Juel1)VDB71164
245 _ _ |a Fast, quantitative in situ hybridization of rare mRNAs using 14C-standards and phosphorus imaging
260 _ _ |a Amsterdam [u.a.]
|b Elsevier Science
|c 2009
300 _ _ |a 56 - 61
336 7 _ |a Journal Article
|0 PUB:(DE-HGF)16
|2 PUB:(DE-HGF)
336 7 _ |a Output Types/Journal article
|2 DataCite
336 7 _ |a Journal Article
|0 0
|2 EndNote
336 7 _ |a ARTICLE
|2 BibTeX
336 7 _ |a JOURNAL_ARTICLE
|2 ORCID
336 7 _ |a article
|2 DRIVER
440 _ 0 |a Journal of Neuroscience Methods
|x 0165-0270
|0 9911
|y 1
|v 185
500 _ _ |a Record converted from VDB: 12.11.2012
520 _ _ |a The 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.
536 _ _ |a Funktion und Dysfunktion des Nervensystems
|c P33
|2 G:(DE-HGF)
|0 G:(DE-Juel1)FUEK409
|x 0
588 _ _ |a Dataset connected to Web of Science, Pubmed
650 _ 2 |2 MeSH
|a Animals
650 _ 2 |2 MeSH
|a Cacodylic Acid: toxicity
650 _ 2 |2 MeSH
|a Carbon Radioisotopes: chemistry
650 _ 2 |2 MeSH
|a Hippocampus: drug effects
650 _ 2 |2 MeSH
|a Hippocampus: metabolism
650 _ 2 |2 MeSH
|a Hippocampus: physiopathology
650 _ 2 |2 MeSH
|a Image Processing, Computer-Assisted: instrumentation
650 _ 2 |2 MeSH
|a Image Processing, Computer-Assisted: methods
650 _ 2 |2 MeSH
|a In Situ Hybridization: methods
650 _ 2 |2 MeSH
|a Male
650 _ 2 |2 MeSH
|a Neurochemistry: methods
650 _ 2 |2 MeSH
|a Oligonucleotides: chemistry
650 _ 2 |2 MeSH
|a Phosphorus Radioisotopes: chemistry
650 _ 2 |2 MeSH
|a RNA, Messenger: analysis
650 _ 2 |2 MeSH
|a RNA, Messenger: metabolism
650 _ 2 |2 MeSH
|a Rats
650 _ 2 |2 MeSH
|a Rats, Wistar
650 _ 2 |2 MeSH
|a Receptors, AMPA: genetics
650 _ 2 |2 MeSH
|a Reference Standards
650 _ 2 |2 MeSH
|a Time Factors
650 _ 7 |0 0
|2 NLM Chemicals
|a Carbon Radioisotopes
650 _ 7 |0 0
|2 NLM Chemicals
|a Oligonucleotides
650 _ 7 |0 0
|2 NLM Chemicals
|a Phosphorus Radioisotopes
650 _ 7 |0 0
|2 NLM Chemicals
|a RNA, Messenger
650 _ 7 |0 0
|2 NLM Chemicals
|a Receptors, AMPA
650 _ 7 |0 0
|2 NLM Chemicals
|a glutamate receptor ionotropic, AMPA 1
650 _ 7 |0 0
|2 NLM Chemicals
|a glutamate receptor ionotropic, AMPA 2
650 _ 7 |0 75-60-5
|2 NLM Chemicals
|a Cacodylic Acid
650 _ 7 |a J
|2 WoSType
653 2 0 |2 Author
|a Quantitative in situ hybridization
653 2 0 |2 Author
|a Standardization
653 2 0 |2 Author
|a Microscales
653 2 0 |2 Author
|a Phosphorus imaging
653 2 0 |2 Author
|a Autoradiography
653 2 0 |2 Author
|a Neurotransmitter receptors
653 2 0 |2 Author
|a Dimethyl arsenic acid
700 1 _ |a Cremer, M.
|b 1
|u FZJ
|0 P:(DE-Juel1)VDB21539
700 1 _ |a Lopez Escobar, J.
|b 2
|u FZJ
|0 P:(DE-Juel1)VDB89117
700 1 _ |a Speckmann, E.J.
|b 3
|0 P:(DE-HGF)0
700 1 _ |a Zilles, K.
|b 4
|u FZJ
|0 P:(DE-Juel1)131714
773 _ _ |a 10.1016/j.jneumeth.2009.09.010
|g Vol. 185, p. 56 - 61
|p 56 - 61
|q 185<56 - 61
|0 PERI:(DE-600)1500499-5
|t Journal of neuroscience methods
|v 185
|y 2009
|x 0165-0270
856 7 _ |u http://dx.doi.org/10.1016/j.jneumeth.2009.09.010
909 C O |o oai:juser.fz-juelich.de:7022
|p VDB
913 1 _ |k P33
|v Funktion und Dysfunktion des Nervensystems
|l Funktion und Dysfunktion des Nervensystems
|b Gesundheit
|0 G:(DE-Juel1)FUEK409
|x 0
914 1 _ |y 2009
915 _ _ |0 StatID:(DE-HGF)0010
|a JCR/ISI refereed
920 1 _ |0 I:(DE-Juel1)INM-2-20090406
|k INM-2
|l Molekulare Organisation des Gehirns
|g INM
|x 0
920 1 _ |0 I:(DE-82)080010_20140620
|k JARA-BRAIN
|l Jülich-Aachen Research Alliance - Translational Brain Medicine
|g JARA
|x 1
970 _ _ |a VDB:(DE-Juel1)115510
980 _ _ |a VDB
980 _ _ |a ConvertedRecord
980 _ _ |a journal
980 _ _ |a I:(DE-Juel1)INM-2-20090406
980 _ _ |a I:(DE-82)080010_20140620
980 _ _ |a UNRESTRICTED
981 _ _ |a I:(DE-Juel1)VDB1046

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