Tissue-specific selection of reference genes is required for expression studies in the mouse model of maternal protein undernutrition

Suboptimal maternal nutrition during gestation results in the establishment of long-term phenotypic changes and an increased disease risk in the offspring. To elucidate how such environmental sensitivity results in physiological outcomes, the molecular characterisation of these offspring has become...

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Bibliographic Details
Published in:Theriogenology 2011-08, Vol.76 (3), p.558-569
Main Authors: Lucas, E.S., Watkins, A.J., Cox, A.L., Marfy-Smith, S.J., Smyth, N., Fleming, T.P.
Format: Article
Language:English
Subjects:
animal models
Animals
blastocyst
Blastocyst - metabolism
embryonic stem cells
Embryonic Stem Cells - metabolism
Female
Gene Expression
Gene Expression Profiling
Gene Expression Regulation, Developmental
genes
heart
Intra-Abdominal Fat - metabolism
Kidney - metabolism
kidneys
liver
Liver - metabolism
malnutrition
Maternal diet
maternal nutrition
Maternal Nutritional Physiological Phenomena
metabolism
Mice
Mouse embryonic stem cells
Mouse embryos
Myocardium - metabolism
placenta
Placenta - metabolism
Pregnancy
progeny
Protein-Energy Malnutrition - genetics
Protein-Energy Malnutrition - metabolism
Real-time qPCR
Reference genes
Reference Values
risk
Sex Factors
yolk sac
Yolk Sac - metabolism
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Summary:Suboptimal maternal nutrition during gestation results in the establishment of long-term phenotypic changes and an increased disease risk in the offspring. To elucidate how such environmental sensitivity results in physiological outcomes, the molecular characterisation of these offspring has become the focus of many studies. However, the likely modification of key cellular processes such as metabolism in response to maternal undernutrition raises the question of whether the genes typically used as reference constants in gene expression studies are suitable controls. Using a mouse model of maternal protein undernutrition, we have investigated the stability of seven commonly used reference genes ( 18s, Hprt1, Pgk1, Ppib, Sdha, Tbp and Tuba1) in a variety of offspring tissues including liver, kidney, heart, retro-peritoneal and inter-scapular fat, extra-embryonic placenta and yolk sac, as well as in the preimplantation blastocyst and blastocyst-derived embryonic stem cells. We find that although the selected reference genes are all highly stable within this system, they show tissue, treatment and sex-specific variation. Furthermore, software-based selection approaches rank reference genes differently and do not always identify genes which differ between conditions. Therefore, we recommend that reference gene selection for gene expression studies should be thoroughly validated for each tissue of interest.
ISSN:0093-691X
1879-3231
DOI:10.1016/j.theriogenology.2011.03.008