The BCPH Unit of Molecular Physiology, Department of Chemistry, Biology and Marine Science, Faculty of Science, University of the Ryukyus, Nishihara, Okinawa 903-0213, Japan.
Cells. 2024 Sep 2;13(17):1476. doi: 10.3390/cells13171476.
One of the important genes for eyespot development in butterfly wings is . Its function has been evaluated via several methods, including CRISPR/Cas9 genome editing. However, functional inhibition may be performed at the right time at the right place using a different method. Here, we used a novel protein delivery method for pupal wing tissues in vivo to inactivate a target protein, Distal-less, with a polyclonal anti-Distal-less antibody using the blue pansy butterfly . We first demonstrated that various antibodies including the anti-Distal-less antibody were delivered to wing epithelial cells in vivo in this species. Treatment with the anti-Distal-less antibody reduced eyespot size, confirming the positive role of Distal-less in eyespot development. The treatment eliminated or deformed a parafocal element, suggesting a positive role of Distal-less in the development of the parafocal element. This result also suggested the integrity of an eyespot and its corresponding parafocal element as the border symmetry system. Taken together, these findings demonstrate that the antibody-mediated protein knockdown method is a useful tool for functional assays of proteins, such as Distal-less, expressed in pupal wing tissues, and that Distal-less functions for eyespots and parafocal elements in butterfly wing color pattern development.
在蝴蝶翅膀的眼斑发育中，一个重要的基因是. 已经通过几种方法评估了其功能，包括 CRISPR/Cas9 基因组编辑。然而，使用不同的方法可以在适当的时间和地点进行功能抑制。在这里，我们使用一种新的蛋白递呈方法，在蓝目菊凤蝶的蛹期翅组织中体内抑制靶蛋白 Distal-less，使用多克隆抗 Distal-less 抗体。我们首先证明，包括抗 Distal-less 抗体在内的各种抗体可以在该物种的翅上皮细胞中体内递呈。用抗 Distal-less 抗体处理会减小眼斑的大小，这证实了 Distal-less 在眼斑发育中的积极作用。这种处理消除或使一个旁焦点元素变形，提示 Distal-less 对旁焦点元素的发育有积极作用。这一结果还表明眼斑及其相应的旁焦点元素作为边界对称系统是完整的。总之，这些发现表明抗体介导的蛋白敲低方法是研究在蛹期翅组织中表达的蛋白（如 Distal-less）功能的有用工具，并且 Distal-less 对蝴蝶翅膀颜色图案发育中的眼斑和旁焦点元素起作用。
