Comprehensive Proteomics Analysis of In Vitro Canine Oviductal Cell-Derived Extracellular Vesicles

Abstract

Dogs (Canis lupus familiaris) have unique and peculiar reproductive characteristics. While the interplay between in vitro oviductal cell-derived extracellular vesicles (OC-EVs) and cumulus-oocyte complexes in dogs has begun to be elucidated, no study has yet provided extensive information on the biological content and physiological function of OC-EVs and their role in canine oocyte development. Here, we aimed to provide the first comprehensive proteomic analysis of OC-EVs. We identified 398 proteins as present in all OC-EVs samples. The functional enrichment analysis using Gene Ontology terms and an Ingenuity Pathway Analysis revealed that the identified proteins were involved in several cellular metabolic processes, including translation, synthesis, expression, and protein metabolism. Notably, the proteins were also involved in critical canonical pathways with essential functions in oocyte and embryo development, such as ERK/MAPK, EIF2, PI3K/AKT, and mTOR signaling. These data would be an important resource for studying canine reproductive physiology and establishing a successful in vitro embryo production system in dogs.

Oviduct epithelial cells‐derived extracellular vesicles improve preimplantation developmental competence of in vitro produced porcine parthenogenetic and cloned embryos

“…EVs act as cell‐to‐cell communication nanoparticles (Mathieu et al, 2019) containing micro RNA and small RNA molecules that mediate cell signaling and epigenetics (Park et al, 2019; Saadeldin et al, 2014; Spadafora, 2020). Results showed the expression of SOD1 in the OEC EVs that supports the evidence that oviductal secretion provide antioxidative defense to the early embryos (Lee et al, 2021; Li & Winuthayanon, 2017). The oviduct is a vital organ for early embryonic development due to the secretion of epithelial cells (Kölle et al, 2020; Lee et al, 2021).…”

“…Oviduct epithelial cells (OESs) secrete oviductal fluid (OF) and play important roles in reproductive events, participating in final gamete maturation, fertilization, and early embryonic development (González‐Brusi et al, 2020; Tukur et al, 2020). Moreover, the OF mediates an early embryo‐maternal dialog through cell‐to‐cell contacts, secreted molecules, and extracellular vesicles (EVs) (Lee et al, 2021; Saint‐Dizier, Schoen, Chen, Banliat, & Mermillod, 2019).…”

“…EVs are present in oviduct fluids and have emerged as a new method of cell‐to‐cell communication (Harris et al, 2020). Oviductal EVs have been identified as crucial components of the OF and potential mediators of embryonic‐maternal interactions (Almiñana & Bauersachs, 2019; Bridi et al, 2020; Lee et al, 2021; Saadeldin et al, 2021, 2014, 2015). EVs as nanoparticles contain diverse bioactive molecules such as proteins, messenger RNAs (mRNAs), small ncRNAs, microRNAs (miRNAs), lipids, amino acids, and growth factors (B. Yang et al, 2019).…”

Porcine oviductal extracellular vesicles facilitate early embryonic development via relief of endoplasmic reticulum stress

“…Intriguingly, melatonin has been identified in OF‐derived EVs from rabbits (Qu, Luo, et al, 2020). Furthermore, a recent study revealed that several proteins associated with rough ER functions, such as endoplasmin (HSP90), 40S ribosomal protein S26, and 60S ribosomal protein L13a, were also detected in in vitro canine oviductal cell‐derived EVs, as determined by liquid chromatography‐tandem mass spectrometry (Lee et al, 2021). In line with the above evidence, we propose that porcine oviductal EVs may relieve ER stress in preimplantation embryos via the bioactive molecules contained in them.…”

Editorial: Biofluid Extracellular Vesicles and Their Involvement in Animal Reproductive Physiology

“…EVs, including exosomes and microvesicles, are membrane-enveloped particles secreted by living cells to the surrounding microenvironment and body fluids, and contain proteins, messenger RNA (mRNA), and microRNAcargoes. In different mammalian species, EVs have been detected in all reproductive biofluids with emerging evidence on their roles in gametogenesis, fertilization, early embryo development, and implantation (2)(3)(4)(5)(6).…”