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Specific Tandem 3'UTR Patterns and Gene Expression Profiles in Mouse Thy1+ Germline Stem Cells
Yan Huang1, Yuanyan Xiong1,2, Zhuoheng Lin1
1Key Laboratory of Reproductive Medicine of Guangdong Province, the First Affiliated Hospital and Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou, 51000, China.
Plos One
|December 30, 2015
Summary
This study reveals alternative polyadenylation (APA) patterns in mouse germline stem cells (GSCs), showing shorter 3' untranslated regions (3'UTRs) and identifying novel genes crucial for GSC function and spermatogenesis.
Area of Science:
- Molecular Biology
- Genomics
- Stem Cell Biology
Background:
- Spermatogonial stem cells (SSCs) are essential for male fertility, but their self-renewal and differentiation mechanisms remain incompletely understood.
- Alternative polyadenylation (APA) is a key post-transcriptional regulatory mechanism influencing gene expression and 3' untranslated region (3'UTR) length.
Purpose of the Study:
- To investigate the genome-wide APA patterns and global gene expression profiles in mouse male germline stem cells (GSCs).
- To identify novel genes and regulatory mechanisms involved in GSC self-renewal and spermatogenesis.
Main Methods:
- Isolation and purification of mouse Thy1+ cells (GSCs) using magnetic-activated cell sorting (MACS).
- Application of the sequencing of alternative polyadenylation (APA) sites (SAPAS) strategy for comprehensive poly(A) site analysis.
- Bioinformatic analysis of poly(A) sites, gene expression, and cis-regulatory elements, with validation via qRT-PCR and APA reporter assays.
Main Results:
- Identified 99,944 poly(A) sites across mouse GSCs, ESCs, and MEFs, with ~40% being newly discovered.
- Observed a global trend of shorter 3'UTR lengths in GSCs compared to MEFs, with 334 genes exhibiting distal-to-proximal APA switching.
- Discovered 1337 highly expressed genes in GSCs consistent with their functional characteristics, and identified potential regulatory roles for C-rich elements.
Conclusions:
- The study elucidates specific APA profiles and gene expression patterns in GSCs, providing novel insights into GSC biology.
- Findings highlight the significance of APA in regulating 3'UTR length and gene expression during spermatogenesis.
- The identified genes and regulatory elements offer potential targets for understanding and manipulating male fertility.
