Dynamics of Structural Barriers and Innate Immune Components during Incubation of the Avian Egg: Critical Interplay between Autonomous Embryonic Development and Maternal Anticipation

Hincke, Maxwell T.; Silva, Mylène da; Guyot, Nicolas; Gautron, Joël; McKee, Marc D.; Guabiraba, Rodrigo; Réhault-Godbert, Sophie

doi:10.1159/000493719

Cited by 53 publications

(49 citation statements)

References 122 publications

(158 reference statements)

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“…The CAM has been exploited as a relatively simple, time-and cost-efficient model to study various cancer related processes including angiogenesis, cancer cell invasion, metastasis formation and tumor progression [10][11][12][13] . One important aspect of this in vivo system is that the chick embryo is immunodeficient up until E18, just prior to hatching 14 , affording the possibility to transplant donor cells and tissue without an adverse host response. In addition, the chick in vivo system is relatively unhindered by ethical requirements prior to E14.…”

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confidence: 99%

CAM-Delam: an in vivo approach to visualize and quantify the delamination and invasion capacity of human cancer cells

Palaniappan

Šlekienė

Jonasson

et al. 2020

Sci Rep

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the development of metastases is the major cause of cancer related death. to develop a standardized method that define the ability of human cancer cells to degrade the basement membrane, e.g. the delamination capacity, is of importance to assess metastatic aggressiveness. We now present the in vivo CAM-Delam assay to visualize and quantify the ability of human cancer cells to delaminate and invade. The method includes seeding cancer cells on the chick chorioallantoic membrane (CAM), followed by the evaluation of cancer-induced delamination and potential invasion within hours to a few days. By testing a range of human cancer cell lines in the cAM-Delam assay, our results show that the delamination capacity can be divided into four categories and used to quantify metastatic aggressiveness. our results emphasize the usefulness of this assay for quantifying delamination capacity as a measurement of metastatic aggressiveness, and in unraveling the molecular mechanisms that regulate delamination, invasion, formation of micro-metastases and modulations of the tumor microenvironment. This method will be useful in both the preclinical and clinical characterization of tumor biopsies, and in the validation of compounds that may improve survival in metastatic cancer.

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confidence: 99%

CAM-Delam: an in vivo approach to visualize and quantify the delamination and invasion capacity of human cancer cells

Palaniappan

Šlekienė

Jonasson

et al. 2020

Sci Rep

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“…is initial high mortality is most probably caused by isolate-dependent abilities to grow and invade embryonic tissues, favoring early systemic infections [67]. Ten-day-old chicken embryos do not have a fully developed adaptive immune system to respond to invading pathogens [69].…”

Section: Discussionmentioning

confidence: 99%

Presence of Virulence Genes inEnterococcusSpecies Isolated from Meat Turkeys in Germany Does Not Correlate with Chicken Embryo Lethality

Maasjost

Lüschow

Kleine

et al. 2019

BioMed Research International

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Virulence-associated traits have frequently been studied in enterococci and are considered to contribute towards the pathogenicity of infections. In the present study, Enterococcus isolates were collected during diagnostic investigations from meat turkeys in Germany. Twenty-eight isolates of three different Enterococcus species were analyzed for five selected putative virulence traits to understand their potential role in the pathogenicity using the chicken embryo lethality assay. Ten E. faecalis, ten E. faecium, and eight E. gallinarum isolates were examined for the presence of common virulence genes and their phenotypic expression, namely, the cytolysin operon, five individual cyl genes (cylLL, cylLS, cylM, cylB, and cylA), gelatinase (gelE), hyaluronidase (hylEfm), aggregation substance (asa1), and enterococcal surface protein (esp). The Enterococcus isolates showed significant species-dependent differences in the presence of genotypic traits (p<0.001 by Fisher’s exact test; Cramer’s V = 0.68). At least one gene and up to three virulence traits were found in E. faecalis, while six E. faecium isolates and one E. gallinarum isolate did not display any virulence-associated pheno- or genotype. More than half of the Enterococcus isolates (n = 15) harbored the gelE gene, but only E. faecalis (n = 10) expressed the gelatinase activity in vitro. The hylEfm gene was found in five E. gallinarum isolates only, while seven isolates showed the hyaluronidase activity in the phenotypic assay. In Cramer’s V statistic, a moderate association was indicated for species (V ≤ 0.35) or genotype (V < 0.43) and the results from the embryo lethality assay, but the differences were not significant. All E. gallinarum isolates were less virulent with mortality rates ranging between 0 and 30%. Two E. faecalis isolates were highly virulent, harboring the whole cyl-operon as well as gelE and asa1 genes. Likewise, one E. faecium isolate caused high embryo mortality but did not harbor any of the investigated virulence genes. For the first time, Enterococcus isolates of three different species collected from diseased turkeys were investigated for their virulence properties in comparison. The results differed markedly between the Enterococcus species, with E. faecalis harboring the majority of investigated genes and virulence traits. However, the genotype did not entirely correlate with the phenotype or the isolates’ virulence potential and pathogenicity for chicken embryos.

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“…Amongst vertebrates, the avian class (Aves) appeared around 91 million years ago and is divided into Paleognathae (ancient birds such as ostrich, rhea and emu) and Neognathae (modern birds such as chicken, turkey or zebra finch) (12). All birds produce eggs with a hardmineral shell composed of CaCO3in the form of calcite, that is critical for development of the embryo within an autonomous chamber (13). In addition to defense against physical aggression, the eggshell protects the egg contents against microbial contamination, regulates water and gaseous exchange, and is a calcium source for embryonic bone calcification (14,15).…”

Section: Introductionmentioning

confidence: 99%

Avian eggshell formation reveals a new paradigm for vertebrate mineralization via vesicular amorphous calcium carbonate

Stapane

Roy

Ezagal

et al. 2020

Journal of Biological Chemistry

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Amorphous calcium carbonate (ACC) is an unstable mineral phase, which is progressively transformed into aragonite or calcite in biomineralization of marine invertebrate shells or avian eggshells, respectively. We have previously proposed a model of vesicular transport to provide stabilized ACC in chicken uterine fluid where eggshell mineralization takes place. Herein, we report further experimental support for this model. We confirmed the presence of extracellular vesicles (EVs) using transmission electron microscopy and showed high levels of mRNA of vesicular markers in the oviduct segments where eggshell mineralization occurs. We also demonstrate that EVs contain ACC in uterine fluid using spectroscopic analysis. Moreover, proteomics and immunofluorescence confirmed the presence of major vesicular, mineralization-specific and eggshell matrix proteins in the uterus and in purified EVs. We propose a comprehensive role for EVs in eggshell mineralization, in which annexins transfer calcium into vesicles and carbonic anhydrase 4 catalyzes the formation of bicarbonate ions (HCO3-), for accumulation of ACC in vesicles. We hypothesize that ACC is stabilized by ovalbumin and/or lysozyme or additional vesicle proteins identified in this study. Finally, EDIL3 and MFGE8 are proposed to serve as guidance molecules to target EVs to the mineralization site. We therefore report for the first-time experimental evidence for the components of vesicular transport to supply ACC in a vertebrate model of biomineralization.

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Dynamics of Structural Barriers and Innate Immune Components during Incubation of the Avian Egg: Critical Interplay between Autonomous Embryonic Development and Maternal Anticipation

Cited by 53 publications

References 122 publications

CAM-Delam: an in vivo approach to visualize and quantify the delamination and invasion capacity of human cancer cells

CAM-Delam: an in vivo approach to visualize and quantify the delamination and invasion capacity of human cancer cells

Presence of Virulence Genes inEnterococcusSpecies Isolated from Meat Turkeys in Germany Does Not Correlate with Chicken Embryo Lethality

Avian eggshell formation reveals a new paradigm for vertebrate mineralization via vesicular amorphous calcium carbonate

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