Fig. 4

mRNA expression of myelin-related genes is significantly decreased in iGluSnFR-expressing 5 dpf zebrafish larvae and adult brains. (a) Logarithmic representation of calibrated normalized relative quantity (CNRQ) values showing mRNA quantification of the myelin genes mbpa: 0.045 ± 0.009 versus 0.152 ± 0.053 green; p = 0.0473; t = 1.984 and plp1b: 0.06 ± 0.01 versus 0.16 ± 0.01; blue; p = 0.0023; t = 4.378 together with the housekeeping genes ß-actin: 0.27 ± 0.06 versus 0.32 ± 0.04; red; p = 0.512; t = 0.6962 and ef1a: 4.23 ± 0.77 versus 3.32 ± 0.40; purple; p = 0.3307; t = 1.058; isolated from iGluSnFR-expressing 5 dpf larvae in comparison to iGluSnFR⁽⁻⁾ siblings, respectively. mRNA levels were normalized against ß-actin and ef1a housekeeping gene expression. Each color (dark for iGluSnFR⁽⁺⁾ and light for iGluSnFr⁽⁻⁾) represents a different gene, each shape represents an independent experiment. Data were analyzed by an unpaired two-tailed t test; * indicates a p-value < 0.05; ** indicates a p-value < 0.01; *** indicates a p-value < 0.001. The housekeeping genes ß-actin and ef1a do not show any significant difference whether iGluSnFR is expressed (+) or not (−). Values show mean ± SEM. N = 4 independent experiments as indicated by different shapes. (b) mRNA expression of myelin-related genes is decreased significantly in iGluSnFR-expressing adult fish brain compared to WT littermate brain. As for the larvae fish in (a) logarithmic representation of CNRQ values showing mRNA quantification of the following genes: mbpa: 0.21 ± 0.03 versus 0.53 ± 0.05; p = 0.0014; t = 4.860 plp1b: 0.05 ± 0.009 versus 0.20 ± 0.01; p < 0.001; t = 8.281; ß-actin: 1.44 ± 0.03 versus 1.33 ± 0.04; p = 0.1041; t = 1.914 and ef1a: 0.69 ± 0.02 versus 0.75 ± 0.02; p = 0.1038; t = 1.916. This time though isolated from iGluSnFR-expressing adult fish brain compared to control WT littermate brains, and not from larvae fish. mRNA levels were normalized against ß-actin and ef1a housekeeping genes expression. Both housekeeping genes do not show any significant difference when iGluSnFR is expressed compared to WT brain controls. Again, each color represents a different gene, each shape represents an independent experiment. Data were evaluated by t test. Values show mean ± SEM. N = 4 independent experiments as indicated by different shapes