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Fig. 5. Microarray analysis. (A) Cluster analysis of genes identified as
differentially expressed in brains from control, nicotine- and ethanol-treated
fish. All Zebrafish data were imported into GeneSpring 6.1, analytical
software for microarray analysis. There was a significant 1.5-fold or greater
change in expression of 545 genes between control and treated animals. Using
these 545 genes an experiment tree was generated using a Spearman correlation.
Subsequently, a gene tree was produced using a Pearson correlation. The
resulting tree is shown. Data are coloured based on how far the gene is over-
or underexpressed (relative to a normalized expression level of 1), in terms
of the standard error of the measurement. The colour bar ranges from +3
to –3. The standard error is based on the standard deviation of
the replicate data for a particular gene and condition. Note that the samples
cluster according to their experimental treatment either control, ethanol or
nicotine treated. (B) Quantitative real-time PCR (Q-RT-PCR) was used to
validate the microarray data. Individual genes with different cellular roles
(see Table 3) were selected for
validation. The four genes selected showed similar expression changes when
assessed by Q-RT-PCR as determined by microarray analysis. EtOH, ethanol; Nic,
nicotine; Cal B, calcineurin B; AMMECR1, Alport syndrome, mental retardation,
midface hyperplasia and elliptocytosis chromosome region; GRIA2a, AMPA
glutamate receptor 2a; pBDZR, peripheral benzodiazepine receptor.
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