Quantitative gene expression analysis in a brain slice model: Influence of temperature and incubation media

We describe the RNA integrity (28S/18S ratio) and the messenger RNA (mRNA) expression of genes encoding glyceraldehyde 3-phosphate dehydrogenase (GAPDH), microtubule-associated serine/threonine kinase 2 (Mast2), and β-actin in cortical brain slices incubated for up to 24 h in Ringer’s solution and D...

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Bibliographic Details
Published in:Analytical biochemistry 2008-07, Vol.378 (1), p.99-101
Main Authors: Dos-Anjos, Severiano, Martínez-Villayandre, Beatriz, Montori, Sheyla, Salas, Alberto, Pérez-García, Carlos César, Fernández-López, Arsenio
Format: Article
Language:English
Subjects:
Animals
Brain - metabolism
Culture Media
Electrochemistry
Gene Expression Regulation - genetics
Male
Models, Genetic
Rats
Rats, Sprague-Dawley
RNA, Messenger - genetics
Temperature
Tissue Culture Techniques - methods
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Summary:We describe the RNA integrity (28S/18S ratio) and the messenger RNA (mRNA) expression of genes encoding glyceraldehyde 3-phosphate dehydrogenase (GAPDH), microtubule-associated serine/threonine kinase 2 (Mast2), and β-actin in cortical brain slices incubated for up to 24 h in Ringer’s solution and Dulbecco’s modified Eagle’s medium (DMEM) at 25 and 37 °C. Our data reveal an optimal temporal working window between 1 and 6 h when slices are incubated in Ringer’s solution at 25 °C that allows experiments related to gene expression dynamics to be performed more suitably than those carried out at 37 °C. In addition, we show that reference gene expression may be modified in dynamic experiments and may compromise studies of gene expression.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2008.03.048