Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies

Abstract If the same pigment is found in different tissues in a body, it is natural to assume that the same metabolic pathways are deployed similarly in each tissue. Here we show that this is not the case for ommochromes, the red and orange pigments found in the eyes and wings of butterflies. We tes...

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Main Authors: Shaun Hong Chuen How, Tirtha Das Banerjee, Antόnia Monteiro
Format: Article
Language:English
Published: Nature Portfolio 2023-06-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-023-36491-9
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author Shaun Hong Chuen How
Tirtha Das Banerjee
Antόnia Monteiro
author_facet Shaun Hong Chuen How
Tirtha Das Banerjee
Antόnia Monteiro
author_sort Shaun Hong Chuen How
collection DOAJ
description Abstract If the same pigment is found in different tissues in a body, it is natural to assume that the same metabolic pathways are deployed similarly in each tissue. Here we show that this is not the case for ommochromes, the red and orange pigments found in the eyes and wings of butterflies. We tested the expression and function of vermilion and cinnabar, two known fly genes in the ommochrome pathway, in the development of pigments in the eyes and in the wings of Bicyclus anynana butterflies, both traits having reddish/orange pigments. By using fluorescent in-situ hybridization (HCR3.0) we localized the expression of vermilion and cinnabar in the cytoplasm of pigment cells in the ommatidia but observed no clear expression for either gene on larval and pupal wings. We then disrupted the function of both genes, using CRISPR-Cas9, which resulted in the loss of pigment in the eyes but not in the wings. Using thin-layer chromatography and UV–vis spectroscopy we identified the presence of ommochrome and ommochrome precursors in the orange wing scales and in the hemolymph of pupae. We conclude that the wings either synthesize ommochromes locally, with yet unidentified enzymes or incorporate these pigments synthesized elsewhere from the hemolymph. Different metabolic pathways or transport mechanisms, thus, lead to the presence of ommochromes in the wings and eyes of B. anynana butterflies.
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spelling doaj.art-8d4b714e1f8a4bd69bdf7fdd1d4c4b5b2023-06-25T11:16:40ZengNature PortfolioScientific Reports2045-23222023-06-0113111210.1038/s41598-023-36491-9Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterfliesShaun Hong Chuen How0Tirtha Das Banerjee1Antόnia Monteiro2Department of Biological Sciences, National University of SingaporeDepartment of Biological Sciences, National University of SingaporeDepartment of Biological Sciences, National University of SingaporeAbstract If the same pigment is found in different tissues in a body, it is natural to assume that the same metabolic pathways are deployed similarly in each tissue. Here we show that this is not the case for ommochromes, the red and orange pigments found in the eyes and wings of butterflies. We tested the expression and function of vermilion and cinnabar, two known fly genes in the ommochrome pathway, in the development of pigments in the eyes and in the wings of Bicyclus anynana butterflies, both traits having reddish/orange pigments. By using fluorescent in-situ hybridization (HCR3.0) we localized the expression of vermilion and cinnabar in the cytoplasm of pigment cells in the ommatidia but observed no clear expression for either gene on larval and pupal wings. We then disrupted the function of both genes, using CRISPR-Cas9, which resulted in the loss of pigment in the eyes but not in the wings. Using thin-layer chromatography and UV–vis spectroscopy we identified the presence of ommochrome and ommochrome precursors in the orange wing scales and in the hemolymph of pupae. We conclude that the wings either synthesize ommochromes locally, with yet unidentified enzymes or incorporate these pigments synthesized elsewhere from the hemolymph. Different metabolic pathways or transport mechanisms, thus, lead to the presence of ommochromes in the wings and eyes of B. anynana butterflies.https://doi.org/10.1038/s41598-023-36491-9
spellingShingle Shaun Hong Chuen How
Tirtha Das Banerjee
Antόnia Monteiro
Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies
Scientific Reports
title Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies
title_full Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies
title_fullStr Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies
title_full_unstemmed Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies
title_short Vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of Bicyclus anynana butterflies
title_sort vermilion and cinnabar are involved in ommochrome pigment biosynthesis in eyes but not wings of bicyclus anynana butterflies
url https://doi.org/10.1038/s41598-023-36491-9
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AT tirthadasbanerjee vermilionandcinnabarareinvolvedinommochromepigmentbiosynthesisineyesbutnotwingsofbicyclusanynanabutterflies
AT antoniamonteiro vermilionandcinnabarareinvolvedinommochromepigmentbiosynthesisineyesbutnotwingsofbicyclusanynanabutterflies