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Analysis of gene expression in the rat hippocampus using Real Time PCR reveals high inter-individual variation in mRNA expression levels
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för genetik och patologi.
Vise andre og tillknytning
2002 (engelsk)Inngår i: Journal of Neuroscience Research, ISSN 0360-4012, E-ISSN 1097-4547, Vol. 67, nr 2, s. 225-34Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

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.

sted, utgiver, år, opplag, sider
2002. Vol. 67, nr 2, s. 225-34
HSV kategori
Identifikatorer
URN: urn:nbn:se:uu:diva-90901DOI: 10.1002/jnr.10105OAI: oai:DiVA.org:uu-90901DiVA, id: diva2:163418
Tilgjengelig fra: 2003-10-14 Laget: 2003-10-14 Sist oppdatert: 2018-01-13bibliografisk kontrollert
Inngår i avhandling
1. High-resolution Studies of mRNA Expression in Brain: A Search for Genes Differently Expressed in Schizophrenia
Åpne denne publikasjonen i ny fane eller vindu >>High-resolution Studies of mRNA Expression in Brain: A Search for Genes Differently Expressed in Schizophrenia
2003 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Gene expression differences between patients and controls can be used to find susceptibility genes and drug targets for a disease. High-resolution strategies are required because the differences between the investigated groups may be small and numerous factors may affect the mRNA quantity. This thesis is based on the use of real-time RT-PCR combined with a new statistical approach, developed to detect small differences between patients and controls and differences due to patient subgroups.

Comparisons between human brain biopsy and autopsy samples showed that post-mortem tissue can be used to make conclusions on the relative mRNA levels in the living brain.

Power analysis based on human brain mRNA expression from 14 genes adjusted with two reference genes, revealed that a sample size of 50 patients and 50 controls was required to detect a 2-fold difference with a power and a confidence of 95%. A similar study in rats revealed that approximately the same sample size was required for rat brain mRNA expression studies.

The mRNA levels of several genes were studied in 55 schizophrenia and 55 control prefrontal brain autopsies, using a novel and more powerful statistical analysis. The serotonin receptor 2C gene (HTR2C) showed a significant 1.5-fold decrease in the patients as compared to controls, and the monoamine oxidase B gene (MAOB) a 1.2-fold increase.

The mechanism behind the decrease of HTR2C mRNA levels was investigated by studying the correlation of drug treatment and HTR2C promoter polymorphisms to the HTR2C expression levels. The observed decrease was present in untreated patients, suggesting that the HTR2C mRNA decrease is correlated with the disease and not the treatment. There was no association between promoter polymorphisms and HTR2C expression levels. Thus, the molecular mechanism for the decreased expression remains unclear. Nevertheless, the results support a role for monoaminergic synapses in schizophrenia.

sted, utgiver, år, opplag, sider
Uppsala: Acta Universitatis Upsaliensis, 2003. s. 47
Serie
Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1104-232X ; 894
Emneord
Genetics, mRNA, gene expression, real-time RT-PCR, schizophrenia, 5-HT (serotonin) receptor 2C, brain, psychiatric genetics, Genetik
HSV kategori
Identifikatorer
urn:nbn:se:uu:diva-3605 (URN)91-554-5755-X (ISBN)
Disputas
2003-11-07, Lindahlsalen, EBC, Uppsala, 09:15 (engelsk)
Opponent
Veileder
Tilgjengelig fra: 2003-10-14 Laget: 2003-10-14 Sist oppdatert: 2018-01-13bibliografisk kontrollert

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