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- Progesterone differentially affects the transcriptomic profiles of cow endometrial cell typesPublication . Pereira, Gonçalo; Guo, YZ; Silva, E; Bevilacqua, C; Charpigny, G; Lopes-da-Costa, Luís; Humblot, PBackground: The endometrium is a heterogeneous tissue composed of luminal epithelial (LE), glandular epithelial (GE), and stromal cells (ST), experiencing progesterone regulated dynamic changes during the estrous cycle. In the cow, this regulation at the transcriptomic level was only evaluated in the whole tissue. This study describes specifc gene expression in the three types of cells isolated from endometrial biopsies following laser capture microdissection and the transcriptome changes induced by progesterone in GE and ST cells. Results: Endometrial LE, GE, and ST cells show specifc transcriptomic profles. Most of the diferentially expressed genes (DEGs) in response to progesterone are cell type-specifc (96%). Genes involved in cell cycle and nuclear divi sion are under-expressed in the presence of progesterone in GE, highlighting the anti-proliferative action of pro gesterone in epithelial cells. Elevated progesterone concentrations are also associated with the under-expression of estrogen receptor 1 (ESR1) in GE and oxytocin receptor (OXTR) in GE and ST cells. In ST cells, transcription factors such as SOX17 and FOXA2, known to regulate uterine epithelial-stromal cross-talk conveying to endometrial receptivity, are over-expressed under progesterone infuence. Conclusions: The results from this study show that progesterone regulates endometrial function in a cell type-spe cifc way, which is independent of the expression of its main receptor PGR. These novel insights into uterine physiol ogy present the cell compartment as the physiological unit rather than the whole tissue.
- Genomic and phenotypic characterization of Campylobacter fetus subsp. venerealis strainsPublication . Silva, Marta Filipa Serra da; Pereira, Ana L.; Fraqueza, M. J.; Pereira, Gonçalo; Mateus, Luisa; Lopes-da-Costa, Luís; Silva, EThe pathogenesis mechanisms of Campylobacter fetus subsp. venerealis (Cfv), the etiologic agent of Bovine Genital Campylobacteriosis remain elusive. This study evaluated the virulence potential and biovar characteristics of Cfv isolates (n = 13) by PCR screening of putative virulencefactor (VF) genes, Multilocus Sequence Typing (MLST) analysis, antimicrobial susceptibility to tetracycline, penicillin, enrofloxacin and streptomycin testing and whole-genome sequencing (WGS; n = 5), also comparing the latter with 26 other whole-genome sequences of Cfv strains. The putative VF genes encoding type IV secretion system of Cfv (virB2-virB11/virD4) were absent in 92% of isolates, including isolates from aborted foetuses, evidencing that these VF genes are not essential for Cfv pathogenicity. The parA gene, used as a Cfv diagnostic molecular target, was detected in only 3 of 13 isolates, invalidating its use for diagnosis purposes. Three novel sequence types were identified by MLST. Although no in vitro antimicrobial resistance was detected, WGS identified antimicrobial resistance-related genes, including those encoding the multidrug efflux pumps CmeABC and YkkCD, indicating that their presence is not enough to provide antimicrobial resistance. The SNP and accessory protein families analysis segregated the Cfv and Cfv biovar intermedius (Cfvi) strains into different clusters. In conclusion, this study evidenced virulence potential and biovar characteristics of Cfv and Cfvi, which are of relevance for the control of Bovine Genital Campylobacteriosis.
- Enzymes present in neutrophil extracellular traps may stimulate the fibrogenic PGF(2 alpha) pathway in the mare endometriumPublication . Rebordão, Maria Rosa; Amaral, Ana; Fernandes, Carina; Silva, E; Lukasik, Karolina; Szóstek-Mioduchowska, Anna; Bravo, Pedro; Galvão, António; Skarzynski, Dariusz J.; Ferreira-Dias, GraçaEndometrosis is a fibrotic disease in mare endometrium whose pathological mechanisms remain obscure. Prostaglandin (PG)F2α, despite modulating reproductive physiological processes, may also provoke local pathological collagen deposition (fibrogenesis). Neutrophil extracellular traps (NETs) released during inflammation have been linked to fibrogenesis in several tissues. We have previously shown that enzymes found in NETs increase in vitro collagen production in mare endometrium. In this study, activation of PGF2α-pathway in equine endometrial explants challenged in vitro by enzymes found in NETs is shown. Our results indicate that both endocrine microenvironment (estrous cycle phase) and healthy or pathological conditions of endometrial tissues play an important role in PGF2α-pathway activation. In the endometrium of the follicular phase, we have observed both high production of PGF2α and/or PGF2α receptor gene transcription under the action of enzymes found in NETs, both conditions associated with fibrogenesis in other tissues. Nevertheless, transcription of the PGF2α receptor gene does not appear to be hormone-dependent, albeit their levels seem to be dependent on endometrial category in the mid-luteal phase. This study suggests that enzymes existing in NETs may instigate changes on PGF2α mediators, which may become an additional mechanism of fibrogenesis in mare endometrium.