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. 2020 Aug 13;10:13757. doi: 10.1038/s41598-020-70735-2

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Retinas of heterozygous and homozygous mtnr1a VIL mutant tadpoles display Mel1a protein expression at intensities similar to WT siblings. Double-label confocal immunohistochemistry (IHC) of (a,b) WT, (c,d) F2 VIL heterozygous, and (e,f) F2 VIL homozygous tadpole head paraffin sections was performed with antibodies to X. laevis Mel1a (red label) and Mel1b (green label) melatonin receptors. Panels on the right are higher magnifications of areas in panels on the left side of the figure. Mel1a receptor fluorescent signal intensity and location appears to be similar in animals of all three genotypes. The Mel1a receptor is expressed in RPE, rod and cone photoreceptors (arrows), and inner retinal neurons. The Mel1a and Mel1b receptors display an expression pattern different from each other, although there are areas in which the red and green labels merge as a yellow label, suggestive of co-expression and/or very close proximity to each other. (g,h) WT retina sections processed for IHC with Mel1b antibodies, but without Mel1a antibodies (No 1° red) display specific immunofluorescence for Mel1b (green), but not for Mel1a (red). RPE retinal pigment epithelium, ROS rod outer segments, ONL outer nuclear layer, INL inner nuclear layer, IPL inner plexiform layer, arrows; rod photoreceptor inner segments. Magnification bars = 20 µm.