Improved low-invasive mRNA electroporation method into immature mouse oocytes visualizes protein dynamics during...

Area of Science:

• Reproductive Biology
• Developmental Biology
• Cell Biology

Background:

• Oocyte quality is crucial for successful reproduction.
• Chromosome segregation errors during meiosis I are a major cause of oocyte quality decline.
• Current methods for introducing mRNA into oocytes are invasive and can impair development.

Purpose of the Study:

• To establish a low-invasive method for mRNA introduction into pre-ovulatory mouse oocytes.
• To overcome technical limitations of existing methods like microinjection.
• To enable visualization of oocyte chromosomal dynamics and assess oocyte quality.

Main Methods:

• Developed a low-invasive electroporation (EP) method for mRNA delivery into germinal vesicle (GV) stage mouse oocytes.
• Optimized EP impedance for efficient mRNA introduction.
• Introduced mRNA encoding enhanced green fluorescent protein (EGFP) and histone H2B-EGFP.

Main Results:

• Achieved efficient mRNA introduction into GV oocytes with a high survival rate (95.0%).
• Histone H2B-EGFP mRNA successfully labeled oocytes without affecting blastocyst development rates.
• Demonstrated the feasibility of visualizing chromosomal dynamics and assessing oocyte maturation.

Conclusions:

• The established EP method is a feasible and efficient technique for mRNA delivery into mouse oocytes.
• This method facilitates the study of chromosomal integrity and oocyte quality determination.
• It offers potential for improved pre-implantation embryo selection and understanding of mammalian oocyte development.