RNA Sequencing Reveals the Involvement of Serum Exosomal MiRNAs in Early Pregnancy in Cattle

Overview

Journal Animals (Basel)

Date 2024 Sep 14

PMID 39272385

Authors

Zhongxiang Ji

Binwu Bao

Yumei Wang

Zhengxing Wang

Yi Yang

Jinrui Xu

Xingping Wang

Zhuoma Luoreng

Affiliations

Soon will be listed here.

Abstract

Low fertility is the main cause of the low productivity in beef cattle and is mainly associated with a lack of conception after fertilization. The establishment of early pregnancy in cattle is a complex physiological process, and embryo implantation is crucial for the successful establishment of pregnancy. Exosomal miRNAs play an important role in regulating mammalian embryo implantation and development. This study used synchronous estrus technology to extract exosomes from bovine serum at 0, 14, and 21 days of early pregnancy and analyzed the expression profile of exosomal miRNAs through RNA-seq technology. We identified 472 miRNA precursor sequences and 367 mature miRNA sequences in the three sample groups, with the majority of the miRNAs having high abundance. Differentially expressed miRNAs (DEmiRNAs) were screened, and 20 DEmiRNAs were obtained. The differential expression analysis results show that compared to day 0, there were 15 DEmiRNAs in the serum on day 14 and 5 on day 21 of pregnancy. Compared to the 14th day of pregnancy, there were eight DEmiRNAs in the serum on the 21st day of pregnancy. Bioinformatics analysis shows that the target genes of DEmiRNAs regulated the signaling pathways closely related to early pregnancy, including the VEGF, NF-κB, and MAPK signaling pathways. In addition, the newly discovered miRNAs were bta-miR-3604, bta-miR-2889, bta-miR-3432a, and bta-miR-409b. These results provide a theoretical reference for screening the molecular markers for early pregnancy establishment and maternal recognition of pregnancy (MRP) in cattle and new ideas for shortening the calving interval in cows.

References

Zhang Y, Wang Q, Wang H, Duan E . Uterine Fluid in Pregnancy: A Biological and Clinical Outlook. Trends Mol Med. 2017; 23(7):604-614. DOI: 10.1016/j.molmed.2017.05.002. View

Langmead B, Trapnell C, Pop M, Salzberg S . Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol. 2009; 10(3):R25. PMC: 2690996. DOI: 10.1186/gb-2009-10-3-r25. View

Markkandan K, Ahn K, Lee D, Kim T, Dang C, Hong S . Profiling and identification of pregnancy-associated circulating microRNAs in dairy cattle. Genes Genomics. 2018; 40(10):1111-1117. DOI: 10.1007/s13258-018-0668-2. View

Klohonatz K, Cameron A, Hergenreder J, Silveira J, Belk A, Veeramachaneni D . Circulating miRNAs as Potential Alternative Cell Signaling Associated with Maternal Recognition of Pregnancy in the Mare. Biol Reprod. 2016; 95(6):124. DOI: 10.1095/biolreprod.116.142935. View

Luo H, Kimura K, Aoki M, Hirako M . Vascular endothelial growth factor (VEGF) promotes the early development of bovine embryo in the presence of cumulus cells. J Vet Med Sci. 2002; 64(11):967-71. DOI: 10.1292/jvms.64.967. View

Mincheva-Nilsson L, Baranov V . The role of placental exosomes in reproduction. Am J Reprod Immunol. 2010; 63(6):520-33. DOI: 10.1111/j.1600-0897.2010.00822.x. View

Wilczynska A, Bushell M . The complexity of miRNA-mediated repression. Cell Death Differ. 2014; 22(1):22-33. PMC: 4262769. DOI: 10.1038/cdd.2014.112. View

Chen C, Ridzon D, Broomer A, Zhou Z, Lee D, Nguyen J . Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005; 33(20):e179. PMC: 1292995. DOI: 10.1093/nar/gni178. View

Wang S, Zhang H, Kou H, Chen X, Lu Y, Li L . Early pregnancy diagnoses based on physiological indexes of dairy cattle: a review. Trop Anim Health Prod. 2020; 52(5):2205-2212. DOI: 10.1007/s11250-020-02230-9. View

10.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

11.

Burns G, Brooks K, Spencer T . Extracellular Vesicles Originate from the Conceptus and Uterus During Early Pregnancy in Sheep. Biol Reprod. 2016; 94(3):56. DOI: 10.1095/biolreprod.115.134973. View

12.

Martins J, Policelli R, Pursley J . Luteolytic effects of cloprostenol sodium in lactating dairy cows treated with G6G/Ovsynch. J Dairy Sci. 2011; 94(6):2806-14. DOI: 10.3168/jds.2010-3650. View

13.

Crabbe M, Gijbels K, Visser A, Craeye D, Walbers S, Pinxteren J . Using miRNA-mRNA Interaction Analysis to Link Biologically Relevant miRNAs to Stem Cell Identity Testing for Next-Generation Culturing Development. Stem Cells Transl Med. 2016; 5(6):709-22. PMC: 4878327. DOI: 10.5966/sctm.2015-0154. View

14.

Liu J, Wu Z, Guo S, Zhang T, Ma X, Jiang K . IFN-τ Attenuates LPS-Induced Endometritis by Restraining HMGB1/NF-κB Activation in bEECs. Inflammation. 2021; 44(4):1478-1489. DOI: 10.1007/s10753-021-01433-y. View

15.

Fan X, Muruganandan S, Shallie P, Dhal S, Petitt M, Nayak N . VEGF Maintains Maternal Vascular Space Homeostasis in the Mouse Placenta through Modulation of Trophoblast Giant Cell Functions. Biomolecules. 2021; 11(7). PMC: 8301892. DOI: 10.3390/biom11071062. View

16.

Hamada K, Oike Y, Takakura N, Ito Y, Jussila L, Dumont D . VEGF-C signaling pathways through VEGFR-2 and VEGFR-3 in vasculoangiogenesis and hematopoiesis. Blood. 2000; 96(12):3793-800. View

17.

Liao X, Li Y, Gao R, Geng Y, Chen X, Liu X . Folate deficiency decreases apoptosis of endometrium decidual cells in pregnant mice via the mitochondrial pathway. Nutrients. 2015; 7(3):1916-32. PMC: 4377890. DOI: 10.3390/nu7031916. View

18.

Zheng C, Zou X, Lin H, Zhao B, Zhang M, Luo C . miRNA-185 regulates the VEGFA signaling pathway in dairy cows with retained fetal membranes. Theriogenology. 2018; 110:116-121. DOI: 10.1016/j.theriogenology.2017.12.050. View

19.

Hambruch N, Haeger J, Dilly M, Pfarrer C . EGF stimulates proliferation in the bovine placental trophoblast cell line F3 via Ras and MAPK. Placenta. 2009; 31(1):67-74. DOI: 10.1016/j.placenta.2009.10.011. View

20.

Kozomara A, Birgaoanu M, Griffiths-Jones S . miRBase: from microRNA sequences to function. Nucleic Acids Res. 2018; 47(D1):D155-D162. PMC: 6323917. DOI: 10.1093/nar/gky1141. View