Validation of internal controls for gene expression analysis in the intestine of rats infected with Hymenolepis diminuta.
The non-invasive parasitic cestode Hymenolepis diminuta induces hypertrophy, hyperplasia and other changes in cell activity in the intestine of rats which are indicated in the expression of mRNA. We have investigated various house-keeping genes (GAPDH, beta-actin, 18S and HPRT) and other internal controls (total RNA/unit biomass, total RNA/unit length of intestine) to validate gene expression in the rat intestine after cestode infection and drug-induced neuromodulation. Variation in GAPDH, beta-actin, 18S and HPRT expression was observed in rat jejunal tissue according to treatment. Total RNA/unit length of intestine was found to be the most suitable internal control for normalizing target gene mRNA expression in both infected and/or drug-induced rat intestine. This normalization method may be applied to studies of gene expression levels in intestinal tissue where hypertrophy, hyperplasia, rapid growth and cell differentiation generally occur.
Department of Biology, York University, 4700 Keele St., Toronto, ONT., Canada M3J 1P3.
Tafazzal
T
Hoque
Meetu
M
Bhogal
Meetu
M
Boghal
Rodney A
RA
Webb
D000818:Animals; D020869:Gene Expression Profiling / Q000592:standards*; D005786:Gene Expression Regulation; D006925:Hymenolepiasis / Q000469:parasitology* / Q000503:physiopathology; D048190:Hymenolepis diminuta / Q000472:pathogenicity*; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000469:parasitology; D008297:Male; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*
D011506:Proteins; D012333:RNA, Messenger