Confocal visualization of opsin and thyroid hormone-related gene expression in 4dpf wildtype and trβ2 GOF transgenic larval retinas treated with T3 or DMSO. (A) Single z-projection of lws2 expression overlaid with dio2, trβ2, and rpe65a labels. White rectangular boxes mark magnified areas in the N: Nasal region (left) and cyan rectangular boxes mark areas in the T: Temporal region (right), both rotated 90° (such that RPE is upper and photoreceptor layer is lower in the insets) and without trβ2 or rpe65a labels. N and T panels reveal the naso-temporal patterning of dio2 in the retinal pigment epithelium (RPE). The second column in A shows a full projection of the dio2 channel, highlighting its naso-temporal gradient in both the RPE and neural retina, where dio2 expression decreases from nasal to temporal regions. (B) Single z-projection of lws (labeling both lws1 and lws2) expression overlaid with dio3b and mct8. Smaller white boxes within the lws1&2 column show regions magnified and rotated 90° within the same column (such that nasal is upper, and temporal is lower in the insets). Mct8 expression extends beyond the neural retina into the RPE, with increased expression in T3-treated groups. Larger white and cyan insets in dio3b and mct8 columns magnify areas highlighted by smaller boxes, showing dio3b dorso-ventral patterning in the photoreceptor cell layer and mct8 upregulation in response to T3 treatment. Scale bar = 50 μm.
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