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Updated: Feb 14, 2026

In situ Protocol for Butterfly Pupal Wings Using Riboprobes
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TRPA1 for Butterfly Eyespot Formation.

Momo Ozaki1, Joji M Otaki1

  • 1The BCPH Unit of Molecular Physiology, Department of Chemistry, Biology and Marine Science, Faculty of Science, University of the Ryukyus, Nishihara, Okinawa 903-0213, Japan.

International Journal of Molecular Sciences
|February 13, 2026
PubMed
Summary
This summary is machine-generated.

Transient Receptor Potential Melastatin 8 (TRPM8) channels are crucial for butterfly eyespot development. Manipulating TRPM8 activity in pupae altered eyespot size and color patterns in adult butterfly wings.

Keywords:
TRPA1butterfly wingcolor pattern formationeyespotorganizer

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Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • Butterfly wing color patterns form through 2D morphogenesis and lateral signaling in pupal tissues.
  • Transient Receptor Potential (TRP) channels are multimodal sensors potentially involved in this developmental process.

Purpose of the Study:

  • To investigate the role of TRP channels, specifically TRPA1, in the development of butterfly wing color patterns, focusing on eyespot formation.

Main Methods:

  • Injected TRPA1 antagonists (AM0902, AP-18) and an agonist (JT010) into pupae of the blue pansy butterfly (Junonia orithya).
  • Analyzed changes in eyespot size (core disk, outer black ring, orange ring) in adult wings.
  • Detected TRPA1 mRNA using RT-PCR in pupal wing tissues.
  • Used an antibody against J. orithya TRPA1 to observe effects on color patterns and wing veins.

Main Results:

  • TRPA1 antagonist AM0902 increased eyespot core disk and outer black ring areas.
  • TRPA1 agonist JT010 decreased eyespot core disk area and increased the orange ring area.
  • TRPA1 mRNA was detected in pupal wing tissues.
  • Antibody injection caused aberrant color patterns and wing vein defects.

Conclusions:

  • TRPA1 is expressed in pupal wing tissue and plays a role in integrating signaling for eyespot size and structure.
  • TRPA1 appears to suppress black pigmentation (core disk, outer ring) and enhance non-black (orange) pigmentation during eyespot development.