A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds
Posted on 2020-03-31 - 03:45
Abstract Background Mature sperm carry thousands of RNAs, including mRNAs, lncRNAs, tRNAs, rRNAs and sncRNAs, though their functional significance is still a matter of debate. Growing evidence suggests that sperm RNAs, especially sncRNAs, are selectively retained during spermiogenesis or specifically transferred during epididymis maturation, and are thus delivered to the oocyte at fertilization, providing resources for embryo development. However , a deep characterization of the sncRNA content of bull sperm and its expression profile across breeds is currently lacking. To fill this gap, we optimized a guanidinium–Trizol total RNA extraction protocol to prepare high-quality RNA from frozen bull sperm collected from 40 representative bulls from six breeds. Deep sequencing was performed (40 M single 50-bp reads per sample) to establish a comprehensive repertoire of cattle sperm sncRNA. Results Our study showed that it comprises mostly piRNAs (26%), rRNA fragments (25%), miRNAs (20%) and tRNA fragments (tsRNA, 14%). We identified 5p-halves as the predominant tsRNA subgroup in bull sperm, originating mostly from Gly and Glu isoacceptors. Our study also increased by ~ 50% the sperm repertoire of known miRNAs and identified 2022 predicted miRNAs. About 20% of sperm miRNAs were located within genomic clusters, expanding the list of known polycistronic pri-miRNA clusters and defining several networks of co-expressed miRNAs. Strikingly, our study highlighted the great diversity of isomiRs, resulting mainly from deletions and non-templated additions (A and U) at the 3p end. Substitutions within miRNA sequence accounted for 40% of isomiRs, with G>A, U>C and C>U substitutions being the most frequent variations. In addition, many sncRNAs were found to be differentially expressed across breeds. Conclusions Our study provides a comprehensive overview of cattle sperm sncRNA, and these findings will pave the way for future work on the role of sncRNAs in embryo development and their relevance as biomarkers of semen fertility.
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Sellem, Eli; Marthey, Sylvain; Rau, Andrea; Jouneau, Luc; Bonnet, Aurelie; Perrier, Jean-Philippe; et al. (2020). A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds. figshare. Collection. https://doi.org/10.6084/m9.figshare.c.4912863.v1
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AUTHORS (12)
ES
Eli Sellem
SM
Sylvain Marthey
AR
Andrea Rau
LJ
Luc Jouneau
AB
Aurelie Bonnet
JP
Jean-Philippe Perrier
SF
Sébastien Fritz
CL
Chrystelle Le Danvic
MB
Mekki Boussaha
HK
Hélène Kiefer
HJ
Hélène Jammes
LS
Laurent Schibler