Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2001-8-27
pubmed:abstractText
Comparative analysis of the regional abundances of the various mRNAs in neural tissues requires the quantitation of target nucleic acid sequences while their tissue distribution is preserved. A quantitative in situ hybridization protocol is presented for the assessment of regional levels of calmodulin (CaM) I, II and III mRNAs in the rat brain. Coronal brain cryostat sections were hybridized with multiple CaM [35S]cRNA probes and co-exposed to an autoradiographic film or storage phosphor screen, together with a membrane-based radioactive standard scale. The membrane scale was calibrated against a brain paste standard scale. Regression analyses of the sensitometric graphs of standard scales corresponding to the autoradiographic film and to the storage phosphor screen were performed by means of exponential (ROD=p(1)(1-exp[-p(2)x])) and linear (LI=ax) functions, respectively (ROD is relative optical density, LI is labeling intensity, and x is radioactivity). The ROD/LI values for the hybridized brain regions were converted into cRNA probe copy numbers (estimations of mRNA copy numbers) through use of the above standard scales. This method was applied to compare the regional abundances of multiple CaM mRNAs in the brains of control, dehydrated, chronic ethanol-treated and ethanol withdrawal-treated animals.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1385-299X
pubmed:author
pubmed:issnType
Print
pubmed:volume
8
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
32-44
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11522526-Animals, pubmed-meshheading:11522526-Autoradiography, pubmed-meshheading:11522526-Brain, pubmed-meshheading:11522526-Calibration, pubmed-meshheading:11522526-Calmodulin, pubmed-meshheading:11522526-Central Nervous System Depressants, pubmed-meshheading:11522526-Dehydration, pubmed-meshheading:11522526-Drug Administration Schedule, pubmed-meshheading:11522526-Ethanol, pubmed-meshheading:11522526-In Situ Hybridization, pubmed-meshheading:11522526-Male, pubmed-meshheading:11522526-Protein Isoforms, pubmed-meshheading:11522526-RNA, Messenger, pubmed-meshheading:11522526-Rats, pubmed-meshheading:11522526-Rats, Sprague-Dawley, pubmed-meshheading:11522526-Signal Processing, Computer-Assisted, pubmed-meshheading:11522526-Time Factors, pubmed-meshheading:11522526-Tissue Distribution
pubmed:year
2001
pubmed:articleTitle
Methods for quantification of in situ hybridization signals obtained by film autoradiography and phosphorimaging applied for estimation of regional levels of calmodulin mRNA classes in the rat brain.
pubmed:affiliation
Department of Zoology and Cell Biology, University of Szeged, 2 Egyetem Street, PO Box 659, H-6722, Szeged, Hungary.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't