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Transcriptome analysis of boar spermatozoa with different freezability using RNA-Seq
L Fraser1, P Brym2, C S Pareek3
1Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, 10-719, Olsztyn, Poland.
Theriogenology
|November 13, 2019
Summary
Semen freezability in boars is linked to specific gene expression. Researchers identified key sperm transcripts, like FOS and BAMBI, that are upregulated in poor freezability ejaculates, offering potential markers for cryo-survival prediction.
Area of Science:
- Animal reproduction and genetics
- Sperm biology and cryopreservation
- Molecular biology and transcriptomics
Background:
- Semen freezability is crucial for boar artificial insemination and genetic preservation.
- Genetic factors and sperm transcript profiles are known to influence semen cryo-survival.
- Understanding these molecular underpinnings can improve boar semen preservation techniques.
Purpose of the Study:
- To compare the transcript profiles of spermatozoa from boars with good and poor semen freezability.
- To identify specific differentially expressed genes (DEGs) associated with variations in semen cryo-survival.
- To explore the potential of sperm-related transcripts as predictive markers for boar semen freezability.
Main Methods:
- RNA-Sequencing (RNA-Seq) was performed on spermatozoa from Polish large white boars classified as good (GSF) and poor (PSF) semen freezability.
- Spermatozoa quality was assessed via post-thaw motility, mitochondrial membrane potential, and plasma/acrosome integrity.
- Differential gene expression analysis was conducted using DESeq2, DESeq, and EdgeR; findings were validated with RT-qPCR and Western blotting.
Main Results:
- Numerous differentially expressed gene (DEG) transcripts were detected between GSF and PSF groups.
- Genes associated with inflammation (FOS), apoptosis (NFATC3), spermatogenesis (BAMBI), and energy metabolism (ND6, ACADM) were predominantly upregulated in PSF ejaculates.
- Validation confirmed that upregulated DEG transcripts in spermatozoa correlate with poor semen freezability.
Conclusions:
- Spermatozoa transcriptome profiling is a valuable approach to understand cryo-survival mechanisms.
- Upregulation of specific transcripts (e.g., FOS, NFATC3, BAMBI, PTPRU, PTPN2, ND6, ACADM) is linked to poor boar semen freezability.
- These identified transcripts represent potential biomarkers for predicting boar semen cryo-survival and guiding breeding strategies.
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