Observing silkworm embryos at the fertilization stage using a tissue clearing reagent.

Area of Science:

• Developmental Biology
• Cell Biology
• Insect Science

Background:

• Silkworm (Bombyx mori) embryogenesis is a key model for lepidopteran development.
• Advancements in biological science enable novel tissue observation methods.
• Traditional methods limit the observation of fine morphological and histological features in early development.

Purpose of the Study:

• To apply advanced tissue transparency technology to silkworm eggs.
• To investigate early embryogenesis and nuclear behavior during fertilization.
• To adapt and utilize the CUBIC method for Bombyx mori egg observation.

Main Methods:

• Utilized a modified CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis) tissue clearing method.
• Employed a water-soluble reagent containing urea and amino alcohol for effective tissue clearing.
• Applied the method to Bombyx mori eggs at the fertilization stage.

Main Results:

• Successfully rendered silkworm eggs transparent using the modified CUBIC method.
• Observed the behavior of the egg nucleus, polar body nuclei, and sperm nuclei during fertilization.
• Captured previously unobserved morphological and histological details of early silkworm development.

Conclusions:

• The modified CUBIC method is effective for visualizing early embryogenesis in silkworm eggs.
• This technique provides new insights into nuclear dynamics during fertilization in Bombyx mori.
• Tissue transparency offers a powerful approach for studying lepidopteran developmental processes.