PSIII-2 - The Microrna Profile of Day 30 Pregnant Ipsilateral or Non-pregnant Bovine Uterus

Monday, June 27, 2022

12:00 PM – 1:15 PM CT

Location: Exhibit Hall A

Presenting Author(s)

EA

Erin M. Amiss

Graduate Research and Teaching Assistant
Animal and Veterinary Sciences, Clemson University, USA
Central, South Carolina, United States

Author(s)

KM

Keelee J. McCarty

Post-Doctoral Research Assistant
Fertility and Genetics, Colorado Center of Reproductive Medicine
Lone Tree, CO, United States
SP

Scott L. Pratt, Ph.D.

Professor
Animal and Veterinary Sciences, Clemson University, USA
Clemson, NC, United States
NL

Nathan M. Long, Ph.D.

Associate Professor
Animal and Veterinary Sciences, Clemson University, USA
Clemson, NC, United States

Abstract: MicroRNAs (miRNAs) have been used as biomarkers for various health issues and physiological states, and could be utilized as a tool to detect pregnancy loss. The objective of this study was to determine the variation in miRNA abundance between pregnant and non-pregnant uteri of cows at day 30 post-insemination. Uterine samples were collected from pregnant (n = 4) and non-pregnant (n = 4) cows, and snap frozen in liquid N2. Isolation of uterine RNA was performed using the mirVANA kit and RNA quantification and quality assessed. Eight libraries were generated (4 biological replicates per group) and subjected to RNAseq procedures and bioinformatic analysis by LC Sciences (Houston, TX, USA). Raw sequence data were analyzed using the ACGT101-miR program (LC Sciences, Houston, TX) removing extraneous sequence information. The remaining sequences ranging in size for 18 to 26 nucleotides were mapped to known bovine miRNA sequences (miRbase 22.1) and the bovine genome. Significant and differentially abundant miRNAs were analyzed using normalized deep sequencing counts and were subjected to ANOVA (P < 0.05). There were 55 significant (P < 0.05) differentially abundant microRNAs of which 35 were upregulated in open samples and 20 were upregulated in pregnant samples. Target prediction was performed using TargetScan for differentially abundant miRNA. Functional enrichment analysis of the miRNA target genes was conducted using the Database for Annotation, Visualization and Integrated Discovery (DAVID 2021). Gene targets were analyzed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The top 5 KEGG pathways across all significant and differentially abundant miRNAs were pathways in cancer, PI3K-Akt signaling pathway, MAPK signaling pathway, human papillomavirus infection, and metabolic pathways. These results show that there is variation in the abundance of miRNAs between pregnant and open cows at day 30 post-insemination.