Microinjection methods for sea urchin eggs and blastomeres

Area of Science:

• Developmental Biology
• Marine Biology
• Molecular Biology

Background:

• Microinjection into sea urchin eggs has advanced due to technical improvements.
• Egg size, elasticity, and toughness vary significantly across sea urchin species.
• Adapting microinjection protocols to specific species is essential for successful manipulation.

Purpose of the Study:

• To provide detailed microinjection methods tailored for different sea urchin species.
• To address the challenges posed by species-specific variations in egg characteristics.
• To facilitate research involving gene manipulation and developmental studies in sea urchins.

Main Methods:

• Detailed protocols for microinjection into eggs of *Hemicentrotus pulcherrimus* (tough eggs).
• Adapted methods for *Temnopleurus reevesii* (slightly weak eggs).
• Standard and modified techniques for *Strongylocentrotus purpuratus* (commonly used species).
• Procedures for co-injection of morpholino anti-sense oligonucleotides and mRNAs.
• Methods for microinjection into sea urchin blastomeres at early developmental stages.

Main Results:

• Successfully optimized microinjection techniques for three distinct sea urchin species.
• Demonstrated adaptability of methods to varying egg properties like toughness and elasticity.
• Established protocols for simultaneous delivery of multiple molecules (morpholinos and mRNAs).
• Provided a reliable method for blastomere microinjection, enabling cell-specific manipulation.

Conclusions:

• Species-specific microinjection protocols are vital for efficient manipulation of sea urchin embryos.
• These refined methods enhance the feasibility of genetic and molecular studies in diverse sea urchin models.
• The described techniques support advanced research in developmental biology and gene function using sea urchins.