Small RNA sequencing of cryopreserved semen from single bull revealed altered miRNAs and piRNAs expression between High- and Low-motile sperm populations

Capra, Emanuele; Turri, Federica; Lazzari, Barbara; Cremonesi, Paola; Gliozzi, T.M.; Fojadelli, Ilaria; Stella, Alessandra; Pizzi, Flavia

doi:10.1186/s12864-016-3394-7

Cited by 133 publications

(150 citation statements)

References 69 publications

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“…In addition, sperm tsRNAs are subjected to numerous RNA modifications and edits that contribute to their stability (for review see [24]). Detailed studies of the sperm miRnome have also been undertaken in several species, including humans [25], mice [26,27], horses [28], pigs [29] and cattle [30,31]. Extensive variations in length and sequence composition of miR-NAs were detected by deep sequencing.…”

mentioning

confidence: 99%

A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds

Sellem¹,

Marthey

Rau

et al. 2020

Epigenetics & Chromatin

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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 sncR-NAs, 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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mentioning

confidence: 99%

A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds

Sellem¹,

Marthey

Rau

et al. 2020

Epigenetics & Chromatin

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“…In the present study, the miR-34a was found in a smaller quantity in a low-efficiency sample in IVEPs. Capra et al (2017) identified 83 miRNAs differentially expressed in the high-and low-motile sperm fractions separated by Precoll from four different bulls. Among these miRNAs were not the three miRNAs identified in the present work.…”

Section: Discussionmentioning

confidence: 99%

Micro-RNAs differentially expressed in comparative analysis of sperm samples with high and low efficiency in the in vitro production of bovine (Bos Taurus) embryos

Silva¹,

Gomes²,

Fujimura³

et al. 2019

Biosci. J.

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Infertility or subfertility in bovine males may be related to spermatic microRNAs (miRNAs), whose function seems to be associated with the regulation of gene expression, degradation or storage of messenger RNAs (mRNAs) for later translation into early embryonic development. Thus, the purpose of this study was to identify differentially expressed miRNAs in semen samples from bulls (Bos taurus) with low and high efficiency in the in vitro embryo production (IVEP) and to evaluate if they can be used as markers of semen efficiency for IVEPs. In order to identify miRNA markers of semen efficiency in the in vitro embryo production, eight semen samples from each animal, one bull with high and two bulls with low efficiency in IVEPs were used to perform the RNAseq technique for miRNAs. Initially the samples were washed with PBS to remove the extender semen and subsequently were submitted to RNA extraction protocols performed according to procedures described by mirVana™ miRNA Isolation Kit. Then, the amplification of the miRNAs was carried out, not to mention the preparation of the library (Ion Total RNA-Seq Kit v2), the PCR emulsion reaction, enrichment, as well as the injection of the sample on the chip by the Ion Chef equipment. The sequencing was done on Ion Proton equipment. The comparison between the samples was established using two methodologies for searching for targets to increase the robustness of the analytical procedure: the miRanda program using as cutoff minimum free energy of the hybridization-20 kcal/Mol, 100% of identity between nucleotides 2 and 8 of the miRNA, and the RNAhybrid program, using as cutoff minimum free energy of hybridization-20 kcal/mol. In sum, 1306 miRNAs were identified in the samples. The bta-miR-380-5p, bta-miR-155, bta-miR-30c and bta-miR-34a genes were identified by the Bioinformatics as being strongly differentially expressed between the groups, indicating that these genes may present themselves as possible efficiency markers. However, it has become clear that there is no single miRNA that marks different types and causes of fertility problems.

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“…Studies have investigated miRNAs through RNA sequencing of low and high motile cryopreserved spermatozoa. It was reported that numbers of miRNAs were expressed differentially in both the populations (Capra et al, ). Furthermore, the role of the particular miRNAs targeted genes has been found in the apoptosis, the integrity of the mitochondrial membrane and alteration in spermatogenesis, representing its role in fertility.…”

Section: Mirna and Spermatogenesismentioning

confidence: 99%

Epigenetics and oxidative stress: A twin‐edged sword in spermatogenesis

et al. 2019

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Spermatogenesis is a series of complex events involving a delicate balance between cell proliferation and cell differentiation. Aggregation of chromatins and epigenetic modifications play a vital role in spermatogenesis via regulation of molecular pathways to maintain testicular homeostasis. These epigenetic mechanisms consist of histone modification, chromatin remodelling, DNA methylation and miRNA, etc., which reportedly are critical players in spermatogenesis. One such mechanism involves regulation of oxidative stress in the male reproductive system. The fact that testicular cells contain plenty of unsaturated fatty acids and undergo division at a high rate makes spermatogenic cells highly susceptible to oxidative insult leading to deleterious effect on spermatozoa, which may culminate in infertility in men. Although the correlation between ROS‐mediated oxidative stress and epigenetic alterations has been indicated, research in this regard is still in infancy. Further, the fact that environmental and life style factors are critical determinants of spermatogenic potential indicates the importance of epigenetic regulation of key molecular events in spermatogenesis. Therefore, the current review aims to discuss the ROS‐induced epigenetic deregulation of the molecular mechanism(s) involved in spermatogenesis.

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Small RNA sequencing of cryopreserved semen from single bull revealed altered miRNAs and piRNAs expression between High- and Low-motile sperm populations

Cited by 133 publications

References 69 publications

A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds

A comprehensive overview of bull sperm-borne small non-coding RNAs and their diversity across breeds

Micro-RNAs differentially expressed in comparative analysis of sperm samples with high and low efficiency in the in vitro production of bovine (Bos Taurus) embryos

Epigenetics and oxidative stress: A twin‐edged sword in spermatogenesis

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