Skip to main content
. 2020 Feb 14;30(2):314–328. doi: 10.1089/thy.2019.0598

FIG. 7.

FIG. 7.

The whole-body content of thyroid hormones and relative expression patterns of thyroid hormone receptor genes at embryonic, larvae, juvenile, and adult stages. (A) Total RNA of pooled embryos or larvae (N = 25–30 at each time point from 24 to 120 hpf) were extracted for RT-qPCR analysis. The relative expression of thyroid hormone receptor genes (thraa, thrab, and thrb) were measured and normalized against the housekeeping gene efla as described in Materials and Methods section. (B) Whole-body content of total l-T4 (TT4) and total l-T3 (TT3) in WT zebrafish were determined from larva to adult at the time point (indicated by the arrows) described in Materials and Methods section. Total RNA of larvae (pooled from 3–7 embryos) or total RNA from skin on the belly of 15-dps larvae, 30-day juveniles or 100-day adults were extracted by RT-qPCR analysis. The relative expression of thyroid hormone receptor genes (thraa, thrab, and thrb) were measured and normalized against the housekeeping gene efla as described in Materials and Methods section. For RT-qPCR, the number of whole larvae at 3 and 7 dpf (N = 25–30 each), skin on the belly were dissected and extracted at 15 dpf (N = 35), at 30 dpf (N = 15), and 100 dpf (N = 10). The data are shown as mean ± SE (n = 3). Whole-body content of TT3 and TT4 are shown as ± SE (n = 3). (C) Whole-body content of TT3 and TT4 were determined by ELISA in WT and homozygous thrab 1-bp insertion (m/m) mutant juvenile at 30 dpf and adult at 100 dpf as described in Materials and Methods section. For 30 dpf juveniles, the number of fish used were in the range of 6–9, or 20–31 per sample, each in triplicates for WT fish and 16–19 or 36 fish per sample, each in triplicates for homozygous thrab 1-bp ins (m/m) mutant fish. For 100 dpf adult fish, the number of fish used were 3 per sample, each in triplicates for WT fish and 8–9 or 7–9 fish per sample, each in triplicates for homozygous thrab 1-bp ins (m/m) mutants. The data are shown mean ± SE (n = 6). Two-tail unpaired t-test, p-adjusted <0.05 was used for statistical analysis. ELISA, enzyme-linked immunosorbent assay; T3, triiodothyronine, ; T4, thyroxine; TT3, total T3; TT4, total T4.