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Mouse Round Spermatid Injection

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Published on: January 26, 2024

Piezo-actuated mouse intracytoplasmic sperm injection (ICSI).

Naoko Yoshida¹, Anthony C F Perry

¹Laboratory of Mammalian Molecular Embryology, RIKEN Center for Developmental Biology, 2-2-3 Minatojima Minamimachi, Chuo-ku, Kobe 650-0047, Japan.

Nature Protocols

|April 5, 2007

Summary

A new protocol for mouse intracytoplasmic sperm injection (ICSI) using piezo-actuated micromanipulation significantly improves oocyte survival rates. This method enables efficient genetic manipulation and offspring production, even from non-motile sperm.

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Area of Science:

Reproductive Biology
Developmental Biology
Genetics

Background:

Mouse oocytes are sensitive to conventional intracytoplasmic sperm injection (ICSI) methods, often resulting in cell death.
Developing robust micromanipulation techniques is crucial for advancing mammalian reproductive technologies and genetic studies.
Existing ICSI protocols pose significant challenges for mouse oocyte viability and subsequent applications.

Purpose of the Study:

To present a detailed, step-by-step protocol for mouse piezo-actuated ICSI suitable for non-specialists.
To demonstrate the utility of piezo ICSI for various reproductive and genetic applications in mice.
To establish a simplified method for mouse strain preservation using cryopreserved non-motile sperm.

Main Methods:

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Mouse Round Spermatid Injection

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Utilized piezo-actuated micromanipulation for precise and rapid microinjection needle movement.

Developed a comprehensive 2-4 hour protocol for mouse intracytoplasmic sperm injection (ICSI).

Applied the protocol to achieve successful oocyte penetration and subsequent embryo development.

Main Results:

Piezo-actuated micromanipulation significantly enhances membrane and matrix penetration during ICSI.
The protocol facilitates high survival rates for mouse metaphase II oocytes post-ICSI.
Successful offspring production was achieved using non-motile sperm, simplifying cryopreservation.

Conclusions:

Mouse piezo ICSI offers a highly effective and accessible method for assisted reproduction in laboratory mice.
This technique serves as a foundation for advanced applications like nuclear transfer cloning and transgenesis.
The protocol streamlines micromanipulation for broader applications in mammalian and potentially non-mammalian species.