PUBLICATIONS
Published: 9 September 2023
Johanna Piibor 1, Andres Waldmann 1 2, Keerthie Dissanayake 3, Aneta Andronowska 4, Marilin Ivask 1 5, Madhusha Prasadani 1, Ants Kavak 1, Suranga Kodithuwakku 1 6, Alireza Fazeli
https://doi.org/10.1016/j.mcpro.2023.100642
Abstract:
Uterine environment is tightly and finely regulated via various signalling pathways mediated through endocrine, exocrine, autocrine, juxtracrine and paracrine mechanisms. In utero signalling processes are paramount for normal and abnormal physiology which involves cell to cell, cells to gametes, cells to embryo and even inter-kingdom communications due to presence of uterine microbiota. Extracellular vesicles (EVs) in the uterine fluid (UF) and their cargo components are known to be mediators of in utero signalling and communications. Interestingly, the changes in UF-EV proteome during the bovine oestrous cycle and the effects of these differentially enriched proteins on embryo development are yet to be fully discovered. In this study, shotgun quantitative proteomics-based mass-spectrometry was employed to compare UF-EV proteomes at day 0, 7 and 16 of the oestrous cycle to understand the oestrous cycle dependant dynamics. Furthermore, different phase UF-EVs were supplemented in embryo cultures to evaluate their impact on embryo development. 159 UF-EV proteins were differentially enriched at different time points indicating the UF-EV proteome is cycle dependent. Overall, many identified pathways are important for normal uterine functions, early embryo development and its nutritional needs, such as antioxidant activity, cell morphology and cycle, cellular homeostasis, cell adhesion and carbohydrate metabolic process. Furthermore, the luteal phase UF-EVs supplementation increased in vitro blastocyst rates from 25.0 ± 5.9% to 41.0 ± 4.0% (P ≤ 0.05). Our findings highlight the importance of bovine UF-EV in uterine communications throughout the oestrous cycle. Interestingly, comparison of hormone synchronized EV-proteomes to natural cycle UF-EVs indicated shift of signalling. Finally, UF-EVs can be used to improve embryo production in vitro.
Published: 25 July 2023
https://doi.org/10.3390/ijms241511924
Abstract:
Synchronized crosstalk between the embryo and endometrium during the periconception period is integral to pregnancy establishment. Increasing evidence suggests that the exchange of extracellular vesicles (EVs) of both embryonic and endometrial origin is a critical component of embryo–maternal communication during peri-implantation. Here, we investigated whether embryonic signals in the form of EVs can modulate the endometrial epithelial cell secretome. Receptive endometrial analog RL95-2 cells were supplemented with trophoblast analog JAr cell-derived EVs, and the secretory protein changes occurring in the RL95-2 cells were analyzed using mass spectrometry. EVs of non-trophoblastic origin (HEK 293 cells) were used as the control EV source to supplement endometrial cells. Trophoblast cell-derived EVs enriched endometrial epithelial cell secretions with proteins that support embryo development, attachment, or implantation, whereas control EVs were unable to induce the same effect. The present study suggests that embryonic signals in the form of EVs may prime receptive endometrial epithelial cells to enrich their secretory proteome with critical proteomic molecules with functional importance for periconception milieu formation.