Stallion sperm transcriptome comprises functionally coherent coding and regulatory RNAs as revealed by microarray analysis and RNA-seq.
Abstract: Mature mammalian sperm contain a complex population of RNAs some of which might regulate spermatogenesis while others probably play a role in fertilization and early development. Due to this limited knowledge, the biological functions of sperm RNAs remain enigmatic. Here we report the first characterization of the global transcriptome of the sperm of fertile stallions. The findings improved understanding of the biological significance of sperm RNAs which in turn will allow the discovery of sperm-based biomarkers for stallion fertility. The stallion sperm transcriptome was interrogated by analyzing sperm and testes RNA on a 21,000-element equine whole-genome oligoarray and by RNA-seq. Microarray analysis revealed 6,761 transcripts in the sperm, of which 165 were sperm-enriched, and 155 were differentially expressed between the sperm and testes. Next, 70 million raw reads were generated by RNA-seq of which 50% could be aligned with the horse reference genome. A total of 19,257 sequence tags were mapped to all horse chromosomes and the mitochondrial genome. The highest density of mapped transcripts was in gene-rich ECA11, 12 and 13, and the lowest in gene-poor ECA9 and X; 7 gene transcripts originated from ECAY. Structural annotation aligned sperm transcripts with 4,504 known horse and/or human genes, rRNAs and 82 miRNAs, whereas 13,354 sequence tags remained anonymous. The data were aligned with selected equine gene models to identify additional exons and splice variants. Gene Ontology annotations showed that sperm transcripts were associated with molecular processes (chemoattractant-activated signal transduction, ion transport) and cellular components (membranes and vesicles) related to known sperm functions at fertilization, while some messenger and micro RNAs might be critical for early development. The findings suggest that the rich repertoire of coding and non-coding RNAs in stallion sperm is not a random remnant from spermatogenesis in testes but a selectively retained and functionally coherent collection of RNAs.
Publication Date: 2013-02-11 PubMed ID: 23409192PubMed Central: PMC3569414DOI: 10.1371/journal.pone.0056535Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study investigates the complex population of RNAs found in mature mammalian or horse sperm and their potential roles in regulating spermatogenesis, fertilization and early developmental stages, ultimately leading to a better understanding of sperm-based biomarkers for horse fertility.
Study Overview
- The research primarily focuses on understanding the RNA composition of stallion sperm using modern scientific methods like microarray analysis and RNA-seq (a method used for analyzing the transcriptome).
- This investigation helps shed light on the possible biological functions of these RNAs and their role in fertility and early development.
Methodology
- The researchers used a 21,000-element equine whole-genome oligoarray to analyze sperm and testes RNA, and also employed RNA-seq for further study.
- The microarray analysis identified 6,761 transcripts in the sperm, of which 165 were sperm-enriched, and 155 were differentially expressed between the sperm and testes.
- A total of 70 million raw reads were generated using RNA-seq of which around 50% could be perfectly aligned with the horse reference genome.
Results
- A total of 19,257 sequence tags were successfully mapped to all horse chromosomes and the mitochondrial genome.
- Structural annotation of the sperm transcripts showed alignment with 4,504 known horse and/or human genes, rRNAs, and 82 miRNAs, but 13,354 sequence tags remained anonymous.
- The study also utilized selected equine gene models to identify additional exons and splice variants. Gene Ontology annotations revealed that sperm transcripts were associated with molecular processes and cellular components linked with known sperm functions during fertilization.
Significance
- Some messenger and micro RNAs were identified that might be critical for early development.
- Overall, the research suggests that the diverse range of coding and non-coding RNAs in stallion sperm are not random remnants from spermatogenesis in testes, but selectively retained and functionally coherent collections of RNAs.
In conclusion, the detailed investigation of the stallion sperm transcriptome in this study helps us better understand the role of RNAs during spermatogenesis, improving our knowledge of fertility processes in stallions. The future exploration of these RNAs could lead to the discovery of new sperm-based biomarkers for stallion fertility.
Cite This Article
APA
Das PJ, McCarthy F, Vishnoi M, Paria N, Gresham C, Li G, Kachroo P, Sudderth AK, Teague S, Love CC, Varner DD, Chowdhary BP, Raudsepp T.
(2013).
Stallion sperm transcriptome comprises functionally coherent coding and regulatory RNAs as revealed by microarray analysis and RNA-seq.
PLoS One, 8(2), e56535.
https://doi.org/10.1371/journal.pone.0056535 Publication
Researcher Affiliations
- Department of Veterinary Integrative Biosciences, Texas A&M University, College Station, Texas, United States of America.
MeSH Terms
- Animals
- Biomarkers / metabolism
- Fertility / genetics
- Horses / genetics
- Horses / physiology
- Humans
- Male
- Oligonucleotide Array Sequence Analysis
- RNA, Messenger / genetics
- RNA, Messenger / metabolism
- Regulatory Sequences, Ribonucleic Acid / genetics
- Sequence Analysis, RNA
- Spermatozoa / metabolism
- Spermatozoa / physiology
- Transcriptome
- Y Chromosome / genetics
Conflict of Interest Statement
The authors have declared that no competing interests exist.
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