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  Indian J Med Microbiol
 

Figure 4: Inhibition of the GABAA-ρ receptor induces gene expression changes consistent with canonical retina regeneration. (A) Retinas from Tg(GFAP:GFP)mi2001 zebrafish expressing GFP in Müller glia were dissected, dissociated, and fluorescence sorted to obtain GFP and GFP+ cells without injection of the gamma aminobutyric acid A receptor (GABAA-ρ receptor) inhibitor TPMPA or 24 hours after 25 nmol TPMPA injection. (B) Quantitative reverse transcription PCR was performed on RNA from GFP+ and GFP pools and fold changes in expression were determined for the indicated mRNAs. Fold changes are displayed as 2–ΔΔCt. The red line indicates no change in expression. Fisher's least significant difference tests were used to analyze fold change expression of the indicated RNAs in TPMPA-injected retinas compared to the uninjected retinas. Error bars are shown as the mean ± SEM. n = 3 biological replicates, each with 3 technical triplicates. ****P = 3.3 × 10–6 (ascl1a); **P = 0.0037 (insm1a); ****P = 5.4 × 10–6 (dkk1b); **P = 0.0056 (let-7a). GFAP: Glial fibrillary acid protein; GFP: green fluorescent protein; TPMPA: (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid.

Figure 4:  Inhibition of the GABA<sub>A</sub>-ρ receptor induces gene expression changes consistent with canonical retina regeneration. 
(A) Retinas from <i>Tg(GFAP:GFP)<sup>mi2001</sup></i> zebrafish expressing GFP in Müller glia were dissected, dissociated, and fluorescence sorted to obtain GFP<sup>–</sup> and GFP<sup>+</sup> cells without injection of the gamma aminobutyric acid A receptor (GABA<sub>A</sub>-ρ receptor) inhibitor TPMPA or 24 hours after 25 nmol TPMPA injection. (B) Quantitative reverse transcription PCR was performed on RNA from GFP<sup>+</sup> and GFP<sup>–</sup> pools and fold changes in expression were determined for the indicated mRNAs. Fold changes are displayed as 2<sup>–ΔΔCt</sup>. The red line indicates no change in expression. Fisher's least significant difference tests were used to analyze fold change expression of the indicated RNAs in TPMPA-injected retinas compared to the uninjected retinas. Error bars are shown as the mean ± SEM. <i>n</i> = 3 biological replicates, each with 3 technical triplicates. ****<i>P</i> = 3.3 × 10<sup>–6</sup> (<i>ascl1a</i>); **<i>P</i> = 0.0037 (<i>insm1a</i>); ****<i>P</i> = 5.4 × 10<sup>–6</sup> (<i>dkk1b</i>); **<i>P</i> = 0.0056 (<i>let-7a</i>). GFAP: Glial fibrillary acid protein; GFP: green fluorescent protein; TPMPA: (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid.