Methods for Staging Pupal Periods and Measurement of Wing Pigmentation of &lt;em&gt;Drosophila guttifera&lt;/em&gt;

Fukutomi, Yuichi; Matsumoto, Keiji; Funayama, Noriko; Koshikawa, Shigeyuki

doi:10.3791/56935-v

Cited by 4 publications

(3 citation statements)

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“…Amplified sequences were then inserted into S3aG [41] with the 5’ end oriented closer to the EGFP sequence and the constructs were then introduced into D. melanogaster , VK00006 (attP on cytogenetic location 19E7) [42] using PhiC31 integrase-mediated transgenesis systems (BestGene, CA). Transgenic flies were then crossed to be homozygous for the introduced sequence and maintained at 25 °C with standard cornmeal/sugar/yeast/agar food [43].…”

Section: Methodsmentioning

confidence: 99%

“…Obtained pupas were taken out from the puparium, had their fat and organs removed in 1 x PBS, and were fixed in 1% paraformaldehyde at 4 °C overnight. Fixed wings were taken out from pupal cuticles in 1 x PBS and observed in the fluorescence field using a BX60 microscope (Olympus) [43]. To examine the EGFP expression pattern on wings, we took photographs under the same conditions of light intensity and exposure.…”

Section: Examination Of the Function Of Modified Cis-regulatory Seque...mentioning

confidence: 99%

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Cis-regulatory evolution that caused change inwinglessexpression pattern associated with wing pigmentation pattern ofDrosophila

Karasawa

Saito

Koshikawa

2023

Preprint

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Genetic mechanisms underlying the acquisition of new traits are an important topic in evolutionary developmental biology. Especially, the co-option of important regulatory genes potentially plays an important role in the gain of new traits. However, how the co-option occurs at the sequence level is still elusive.Drosophila guttiferahas a unique wing pigmentation pattern and this is newly gained via the evolution of the expression pattern ofwingless, which induces the pigmentation pattern formation. In this study, to reveal the changes in thecis-regulatory sequence which caused the co-option ofwinglessthat lead to the expression in a new place, we conducted transgenic EGFP reporter assays of alteredcis-regulatory sequences. As a result, the sequence was divided into regions needed to activate expression in the entire wing veins and a region required for repressing expression in excess parts. Comparisons with the homologous sequence ofDrosophila melanogastershowed that the repressive function of thecis-regulatory region is also possessed byD. melanogasterwhile the activating function is newly gained in a lineage leading toD. guttifera. Furthermore, a putative binding site of SMAD transcription factors is shown to be essential for activating expression but also existing in the homologous region ofD. melanogaster. Our results suggest that the pre-existing regulatory sequences in thecis-regulatory region coordinate with the newly gained sequences to acquire the new expression pattern ofwingless.Graphical Abstract

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Section: Methodsmentioning

confidence: 99%

Section: Examination Of the Function Of Modified Cis-regulatory Seque...mentioning

confidence: 99%

Cis-regulatory evolution that caused change inwinglessexpression pattern associated with wing pigmentation pattern ofDrosophila

Karasawa

Saito

Koshikawa

2023

Preprint

Self Cite

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show abstract

“…Amplified sequences were then inserted into S3aG [42] with the 5′ end oriented closer to the EGFP sequence and the constructs were then introduced into D. melanogaster , VK00006 (attP on cytogenetic location 19E7) [43] using PhiC31 integrase‐mediated transgenesis systems (BestGene, CA). Transgenic flies were then crossed to be homozygous for the introduced sequence and maintained at 25 °C with standard cornmeal/sugar/yeast/agar food [44].…”

Section: Methodsmentioning

confidence: 99%

Cis‐regulatory evolution underlying the changes in wingless expression pattern associated with wing pigmentation of Drosophila

2023

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The co‐option of regulatory genes has the potential to play an important role in the evolutionary gain of new traits. However, the changes at the sequence level that underlie such a co‐option event are still elusive. We identified the changes in the cis‐regulatory sequence of wingless that caused co‐option of wingless and led to its expression in new places in Drosophila guttifera, which has unique pigmentation patterns on its wings. The newly gained function of gene expression activation was acquired evolutionarily via a combination of pre‐existing sequences containing a putative binding site for SMAD transcription factors that exhibit an ancestral function in driving expression at crossveins, and a sequence that is specific to the lineage leading to D. guttifera.

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Protocol for dissecting Drosophila pupae and visualizing RNA expression using hybridization chain reaction

Bayala,

Sinha,

Wittkopp

2024

STAR Protocols

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Methods for Staging Pupal Periods and Measurement of Wing Pigmentation of <em>Drosophila guttifera</em>

Cited by 4 publications

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Cis-regulatory evolution that caused change inwinglessexpression pattern associated with wing pigmentation pattern ofDrosophila

Cis-regulatory evolution that caused change inwinglessexpression pattern associated with wing pigmentation pattern ofDrosophila

Cis‐regulatory evolution underlying the changes in wingless expression pattern associated with wing pigmentation of Drosophila

Protocol for dissecting Drosophila pupae and visualizing RNA expression using hybridization chain reaction

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