Analysis of gene expression in the rat hippocampus using Real Time PCR reveals high inter-individual variation in mRNA expression levels.
In mammals, gene transcription is a step subjected to tight regulation mechanisms. In fact, changes in mRNA levels in the central nervous system (CNS) can account for numerous phenotypic differences in brain function. We performed a high-resolution analysis of mRNA expression levels for 37 genes selected from a normal rat hippocampus cDNA library. mRNA amounts were quantified using a Real Time PCR SYBR Green assay. We found that, in general, individuals from an inbred rat population (n = 20) have shown 2-3 times differences in the basal level of expression of the genes analyzed. Up to several fold differences among individuals were observed for certain genes. These inter-individual differences were obtained after correction for the different amounts of mRNA in each sample. Power calculations were performed to determine the number of individuals required to detect reliable differences in expression levels between a control and an experimental group. These data indicated that, depending on the variability of the candidate gene selected, it was necessary to analyze from five to 135 individuals in each group to detect differences of 50% in the levels of mRNA expression between two groups investigated. The comparison of mRNA abundance from different genes revealed a wide range of expression levels for the 37 genes, showing a 26,000-fold difference between the highest and lowest expressed gene.
Alfonso
Julieta
J
Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús-Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de General San Martín, Buenos Aires, Argentina.
Pollevick
Guido D
GD
Castensson
Anja
A
Jazin
Elena
E
Frasch
Alberto C C
AC
D000818:Animals; D052160:Benzothiazoles; D003199:Computer Systems; D018076:DNA, Complementary / Q000032:analysis; D003959:Diamines; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation / Q000502:physiology*; D015723:Gene Library; D014644:Genetic Variation / Q000235:genetics*; D006624:Hippocampus / Q000378:metabolism*; D008297:Male; D023421:Models, Animal; D009419:Nerve Tissue Proteins / Q000235:genetics; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
D052160:Benzothiazoles; D018076:DNA, Complementary; D003959:Diamines; D005456:Fluorescent Dyes; D009419:Nerve Tissue Proteins; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; C098022:SYBR Green I