Helies, V. | Search bioRxiv

bioRxiv2026

Study of holobiont resilience following a break in symbiotic transmission in two rabbit lines selected for feed efficiency

This study was carried out on two rabbit lines, each selected for a feed efficiency criterion (growth on restricted feed for the AlimR line or residual consumption on an adlibitum diet for the ConsR line), introduced into a facility by adoption of newborn kits by SPF (specific pathogen free) females. Following this break in symbiotic transmission, fecal samples from females were used to study the microbiota composition of the two lines over seven generations (17 females per line and per generation). The proportion of Firmicutes was significantly higher in the AlimR line than in the ConsR line (82.22% vs 78.21%, p<0.0001). Conversely, the proportion of Bacteroidota was significantly higher in the ConsR line than in the AlimR line (20.38% vs 16.50%, p<0.0001). The effect of generation was also significant for the abundance of Firmicutes and Bacteroidota (p=0.0009 and p<0.0001, respectively). The Shannon diversity index was significantly higher in the AlimR line than in the ConsR line (5.23 vs 5.14, p=0.02). The effect of generation was also significant for the Shannon index (p=0.003). Finally, a discriminant principal component analysis of the microbiota composition allowed us to distinguish both the two lines and the generations, in particular the 11th selected generation (animals adopted by SPF females) and the 18th generation.

microbiology↗

bioRxiv2020

Reconstitution of intestinal stem cell niche in vitro with pharmacological inhibitors or L-WRN conditioned medium differentially regulates epithelial proliferation, differentiation and barrier function in rabbit caecum organoids

Intestinal organoids are self-organized 3-dimensional (3D) structures formed by a single layer of polarized epithelial cells. This innovative in vitro model is highly relevant to study physiology of the intestinal epithelium and its role in nutrition and barrier function. However, this model has never been developed in rabbits, while it would have potential applications for biomedical and veterinary research. Here, we cultured rabbit caecum organoids with either pharmacological inhibitors (2Ki medium) or L-WRN cells conditioned medium (L-WRN CM) to reconstitute the intestinal stem cell niche in vitro. Large spherical organoids were obtained with the 2Ki medium and this morphology was associated with a high level of proliferation and stem cells markers gene expression. In contrast, organoids cultured with L-WRN CM had a smaller diameter; a greater cell height and part of them were not spherical. When the L-WRN CM was used at low concentration (5%) for two days, the gene expression of stem cells and proliferation markers were very low, while absorptive and secretory cells markers and antimicrobial peptides were elevated. Epithelial cells within organoids were polarized in 3D cultures with 2Ki medium or L-WRN CM (apical side towards the lumen). We cultured dissociated organoid cells in 2D monolayers, which allowed accessibility to the apical compartment. Under these conditions, actin stress fibers were observed with the 2Ki medium, while perijonctionnal localization of actin was observed with the L-WRN CM suggesting, in 2D cultures as well, a higher differentiation level in the presence of L-WRN CM. In conclusion, rabbit caecum organoids cultured with the 2Ki medium were more proliferative and less differentiated than organoids cultured with L-WRN CM. We propose that organoids cultured with the 2Ki medium could be used to rapidly generate in vitro a large number of rabbit intestinal epithelial stem cells while organoids cultured with the L-WRN CM represent a suitable model to study differentiated rabbit epithelium.

cell biology↗