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. 2020 Dec 8;10:21450. doi: 10.1038/s41598-020-78452-6

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© The Author(s) 2020

Open Access This 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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Specific expression of Arhgef33 in the parafovea of the P14 zebra finch retina. (a) In situ hybridization analysis of Arhgef33 expression in the zebra finch retina at P14. The arrowhead indicates a foveal pit. Scale bar = 200 μm. (b) Higher magnification images of the fovea (center) and parafovea (left, left center, right center, right). The Arhgef33 expression was observed in the middle layer of the INL in the parafovea, while the Arhgef33 expression was absent at the center of the fovea. Scale bar = 50 μm. ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. (c) Immunostaining of P14 zebra finch retinal sections using an anti-Sox9 antibody (green) and DAPI (blue). Sox9 signal was detected in the middle layer of the INL. Scale bar = 50 μm. (d) Expression levels of Rhodopsin (left panel), Cone opsin (middle panel), and Arhgef33 (right panel) were measured by RT-qPCR using RNAs purified from the foveal, nasal, and temporal regions of P14 zebra finch retinas. Rhodopsin expression was up-regulated in the nasal and temporal regions, while Cone opsin expression levels were not significantly different between each region. Arhgef33 expression was up-regulated in the foveal region. Significant differences assessed using a Mann–Whitney U test are marked by an asterisk (n = 5; one retina of each bird). n.s., not significant.