A novel high throughput screen to identify candidate molecular networks that regulate spermatogenic stem cell...

Area of Science:

• Reproductive Biology
• Molecular Genetics
• Cellular Biology

Background:

• Male fertility depends on spermatogenic regeneration, driven by undifferentiated spermatogonia.
• Understanding the molecular mechanisms regulating spermatogonial function is critical for fertility research.

Purpose of the Study:

• To develop a high-throughput screening platform for assessing spermatogonial functional capacity.
• To identify novel molecular regulators of spermatogonial regenerative functions.

Main Methods:

• Primary mouse spermatogonial cultures were used for a high-throughput screening platform.
• A large-scale RNAi screen of transcription factors was conducted.
• Pathway analysis, single-cell RNA sequencing, and Zscan2 knockout mouse models were employed.

Main Results:

• A SRCAP-CREBBP/EP300 complex was identified, mediating histone acetylation levels in spermatogonia.
• The previously undescribed transcription factor ZSCAN2 was found to be regulated by this complex and influences self-renewal genes.
• ZSCAN2 deficiency impaired spermatogonial regeneration and affected processes like translation and chromatin modification.

Conclusions:

• The high-throughput screening approach is validated as a tool for uncovering mechanisms in spermatogonial function.
• ZSCAN2 plays a significant role in spermatogonial regenerative capacity, particularly after cytotoxic damage.
• The generated transcription factor database offers insights into male fertility regulation.