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ARS Home » Plains Area » Clay Center, Nebraska » U.S. Meat Animal Research Center » Livestock Bio-Systems » Research » Publications at this Location » Publication #372238

Research Project: Improving Livestock Production by Developing Reproductive and Precision Management Technologies

Location: Livestock Bio-Systems

Title: Comparison of microRNA between ejaculated and epididymal semen

Author
item MENEGATTI ZOCA, SAULO - South Dakota State University
item GE, XIJIN - South Dakota State University
item PERKINS, STEPHANIE - South Dakota State University
item Cushman, Robert - Bob
item McDaneld, Tara
item Keel, Brittney
item PERRY, GEORGE - South Dakota State University

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 3/3/2020
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: The objective of this study was to evaluate microRNA (miRNA) expression from sperm that were ejaculated vs. collected from the epididymis and from the seminal plasma vs. epidydimal fluid. Semen from six sexually mature Angus bulls (~4 years old) with a history of successful breeding were collected by electro-ejaculation weekly for three weeks. Animals were allowed to rest for six weeks before slaughter. Epididymides were harvested and semen from the tail of the epididymis was collected. Sperm and fluids (seminal plasma and epididymal fluid) were separated by centrifugation. Small RNAs (sRNAs) were extracted from both sperm and fluids. Libraries were prepared using the Illumina TruSeq Small RNA preparation kit and sequenced on an Illumina MiSeq. Libraries were uploaded into CLC Genomics Workbench (v12.0.2) and analyzed following the small RNA analysis procedures. Briefly, the sequences were extracted, trimmed and sequences without an adaptor were kept and counted. Sequences were annotated using miRbase and Bos Taurus non-coding RNA database from ENSEMBL (Bos_taurus.ARS-UCD1.2.ncrna). The empirical analysis of differential gene expression was used to compare annotated sRNAs present in the ejaculated vs. epididymal sperm and seminal plasma vs. epididymal fluid; sequences were considered differentially expressed when FDR=0.05 and fold change cutoff=2. In the sperm, there were 3,720 sRNAs identified and 526 were differentially expressed; 217 sRNAs were up-regulated and 309 were down-regulated in the ejaculated vs. epididymal sperm. However, when only known miRNA were analyzed there were 593 miRNAs with 26 differentially expressed and down-regulated in ejaculated vs. epididymal sperm. In the fluids, there were 2,412 sRNAs detected and 404 differentially expressed; 106 sRNAs were up-regulated and 298 down-regulated in seminal plasma vs. epididymal fluid. However, when only known miRNA were analyzed there were 484 miRNAs with 33 that were differentially expressed and down-regulated in seminal plasma vs. epididymal fluid. Of the 38 unique miRNAs differentially expressed, 21 were similar between sperm and fluids, 5 miRNAs were only in the sperm, and 12 miRNAs were only in the fluids. In conclusion, miRNA expression was down-regulated or unchanged in ejaculated compared to epididymal sperm and fluids.