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Home / Equine Research Bank / Andrology / Identification and quantification of coding and long non-cod...
Andrology2020; 8(5); 1409-1418; doi: 10.1111/andr.12791

Identification and quantification of coding and long non-coding RNAs in stallion spermatozoa separated by density.

Abstract: It is not unusual for stallions to have fertility problems. For many, artificial insemination with more dense spermatozoa (isolated by density gradient centrifugation) results in greater pregnancy rates compared with the rates when using unfractionated spermatozoa. RNAs in spermatozoa delivered to the oocyte at conception are required for embryo development. Novel molecular assays of spermatozoa that reflect function are needed to predict the fertility of stallions. To describe and compare the RNA populations in more dense and less dense spermatozoa from stallions. Spermatozoa from five stallions were separated into more dense and less dense populations by density gradient centrifugation. Complementary DNA libraries were made from each of the ten total RNA samples after ribosomal RNA removal. Next-generation sequencing characterized the RNA populations in more and less dense spermatozoa. Quantitative reverse transcription-PCR was used to confirm differential expression of selected RNAs. Stallion spermatozoa contain 11 215 RNAs, with the most prevalent RNA being a 1492 base long non-coding RNA. The levels of 159 RNAs were greater in more dense spermatozoa, while levels of seven other RNAs were greater in less dense spermatozoa. Quantitative reverse transcription-PCR confirmed the threefold greater levels of solute carrier family 26 member 8 (SLC26A8) mRNA in less dense spermatozoa, and sixfold and threefold greater expression levels of the SCP2 sterol binding domain containing 1 (SCP2D1) and spermatogenesis-associated protein 31D1 (SPATA31D1) mRNAs in more dense spermatozoa, respectively. We identified 11 215 RNAs in stallion spermatozoa and 166 with differential expression between more dense and less dense fractions. Many prevalent RNAs were also found in bull, boar, and human spermatozoa. Many differentially expressed RNAs are known to be testis- or spermatozoa-specific. Our results may lead to identification of an RNA population in spermatozoa that is optimal for establishing successful pregnancies.
© 2020 American Society of Andrology and European Academy of Andrology.
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Publication Date: 2020-04-17 PubMed ID: 32243084DOI: 10.1111/andr.12791Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study investigates the types and amounts of RNA in sperm cells from stallions, with the aim to possibly identify key aspects that may help improve fertility.

Objective of the Research

  • The main objective of this research was to identify and compare the RNA populations in sperm cells from stallions, particularly focusing on the difference in RNA presence between denser and less dense spermatozoa.
  • The scientists behind this work were looking to deepen our understanding of fertility and conception in horses.

Methods

  • Studying the sperm from five different stallions, the samples were divided based on their density.
  • After removing ribosomal RNA, complementary DNA libraries were created from each sample. These were then studied through next-generation sequencing, a method optimized for characterizing various RNA populations.
  • The researchers then used quantitative reverse transcription-PCR to confirm the varied levels of certain types of RNAs in dense and less dense spermatozoa.

Findings

  • Through their research, they identified a whopping 11,215 types of RNAs in stallion sperm. They found that the most common type was a 1492 base long non-coding RNA.
  • The scientists discovered that the levels of 159 RNAs were greater in denser sperm cells, while seven other RNAs were more present in less dense spermatozoa.
  • The data further corroborated this when they found greater expression levels of certain RNAs like SCP2 and SPATA31D1 in the denser sperm cells, and higher quantities of SLC26A8 RNA in the less dense spermatozoa.

Significance of the Findings

  • These findings indicate that there are differences in RNA levels between denser and less dense sperm cells, which may be connected to fertility.
  • Understanding the role and behavior of these RNAs could potentially pave the way for developments in fertility treatments or even in the selection processes of artificial insemination procedures.
  • The study further revealed that a lot of the RNAs identified in this research on stallion sperm cells were also found in bull, boar, and even human sperm, which opens many research paths regarding our broader understanding of fertility across mammal species.

Cite This Article

APA
Ing NH, Konganti K, Ghaffari N, Johnson CD, Forrest DW, Love CC, Varner DD. (2020). Identification and quantification of coding and long non-coding RNAs in stallion spermatozoa separated by density. Andrology, 8(5), 1409-1418. https://doi.org/10.1111/andr.12791

Publication

Andrology
ISSN: 2047-2927
NlmUniqueID: 101585129
Country: England
Language: English
Volume: 8
Issue: 5
Pages: 1409-1418

Researcher Affiliations

Ing, Nancy H
  • Department of Animal Science, Texas A&M University, College Station, TX, USA.
Konganti, Kranti
  • Texas A&M Institute for Genome Sciences and Society, Texas A&M University, College Station, TX, USA.
Ghaffari, Noushin
  • AgriLife Genomics and Bioinformatics, Texas A&M University, College Station, TX, USA.
  • Roy G. Perry College of Engineering, Prairie View A&M University, Prairie View, TX, USA.
Johnson, Charles D
  • AgriLife Genomics and Bioinformatics, Texas A&M University, College Station, TX, USA.
Forrest, David W
  • Department of Animal Science, Texas A&M University, College Station, TX, USA.
Love, Charles C
  • Large Animal Clinical Sciences, Texas A&M University, College Station, TX, USA.
Varner, Dickson D
  • Large Animal Clinical Sciences, Texas A&M University, College Station, TX, USA.

MeSH Terms

  • Animals
  • Cattle
  • Centrifugation, Density Gradient
  • Fertility / genetics
  • Horses
  • Humans
  • Male
  • RNA, Long Noncoding / analysis
  • RNA, Long Noncoding / metabolism
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • Spermatozoa / metabolism
  • Swine

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Citations

This article has been cited 3 times.
  1. Ohrt MM, Ing NH. Supplementary L-arginine can enhance reproductive parameters and outcomes in large mammals. Front Vet Sci 2025;12:1740399.
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  2. Sahoo B, Choudhary RK, Sharma P, Choudhary S, Gupta MK. Significance and Relevance of Spermatozoal RNAs to Male Fertility in Livestock. Front Genet 2021;12:768196.
    doi: 10.3389/fgene.2021.768196pubmed: 34956322google scholar: lookup
  3. Zhao S, Heng N, Weldegebriall Sahlu B, Wang H, Zhu H. Long Noncoding RNAs: Recent Insights into Their Role in Male Infertility and Their Potential as Biomarkers and Therapeutic Targets. Int J Mol Sci 2021 Dec 18;22(24).
    doi: 10.3390/ijms222413579pubmed: 34948376google scholar: lookup
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