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Research Project: Validate Causative Mutations in Agriculturally-Important Vertebrates

Location: Plant Genetics Research

Title: Expression of tripartite motif family-like protein 1 and 2 in early conceptus development and placentation in the pig

Author
item EITEL, EMILY - University Of Missouri
item SPONCHIADO, MARIANA - University Of Missouri
item SULLIVAN, RILEY - University Of Missouri
item LUCAS, CAROLINE - University Of Missouri
item Redel, Bethany
item Chen, Paula
item WELLS, KEVIN - University Of Missouri
item PRATHER, RANDALL - University Of Missouri
item WARREN, WESLEY - University Of Missouri
item GEISERT, ROBERT - University Of Missouri

Submitted to: Journal of Reproduction and Fertility
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/27/2025
Publication Date: 4/8/2025
Citation: Eitel, E.K., Sponchiado, M., Sullivan, R.M., Lucas, C.G., Redel, B.K., Chen, P.R., Wells, K.D., Prather, R.S., Warren, W.C., Geisert, R.D. 2025. Expression of tripartite motif family-like protein 1 and 2 in early conceptus development and placentation in the pig. Journal of Reproduction and Fertility. 6(2). https://doi.org/10.1530/RAF-24-0107.
DOI: https://doi.org/10.1530/RAF-24-0107

Interpretive Summary: The biological processes that allow pregnancy establishment, placental attachment, and pregnancy maintenance in the pig are complex and are not completely known. A family of proteins known as the Tripartite Motif Family-Like Proteins (TRIMLs) are associated with the down regulation of inflammation and may be important for allowing the attachment of the developing placenta to the maternal endometrium and ultimately a successful pregnancy. The goal of this study was to identify what roles two of these family members, TRIML1 and TRIML2, which have been identified in gonads, embryos, and placental tissues in humans and mice previously, play in placental attachment and pregnancy maintenance in the pig. By deleting these genes in cells and then using these gene edited cells to produce cloned embryos and then transferring these embryos into a recipient pig, it was found that neither gene affected early establishment and maintenance of pregnancy in the pig, ruling them out as players involved in early pregnancy establishment.

Technical Abstract: The eutherian placenta is highly complex, evolving to regulate the inflammatory phase of pregnancy during conceptus attachment and placental tissue development. Tripartite motif family-like (TRIMLs) proteins are implicated in downregulating inflammation. In mammals, TRIML1 and TRIML2 show preferential expression in gonads, preimplantation embryos and placenta. TRIML1 domains differ between eutherians and marsupials, while TRIML2 is absent in marsupials, suggesting it may play a unique role in regulating the inflammatory phase during conceptus attachment, critical for establishing and maintaining pregnancy to term. This study aimed to investigate the expression pattern of TRIML1 and TRIML2 in various tissues, as well as during embryo development, conceptus attachment, and placental formation in pigs. Transcripts for TRIML2 were detected in embryos, conceptuses, extraembryonic membranes, ovary and testis but not in any of the other tissues examined. In contrast, TRIML1 expression was only observed in testis. In situ hybridization of TRIML1 and TRIML2 confirmed these results. The specific expression of TRIML2 in immune privileged sites is consistent with it serving as an anti-inflammatory factor to provide immunological protection of the eutherian placenta. To further investigate the role of TRIML2, CRISPR/Cas9 gene editing was employed to knock out either TRIML1 (control) or TRIML2. TRIML1/ and TRIML2/ porcine fetal fibroblasts were used for somatic cell nuclear transfer, and the resulting embryos were transferred into surrogate gilts. Early conceptus and placental development were not affected by the loss of conceptus TRIML2. Although a tissue-specific expression pattern was found, TRIML1 or TRIML2 are not required for pregnancy establishment in the pig.