IAL   21557
INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Development of a lactobacilli-drosophyla gnotobiotic model to analyze the impact of prokariotyc glycogen metabolism on the host-microbiota interaction
Autor/es:
ASENCION DIEZ MD; DEKANTY A; CANAL MV
Lugar:
San Miguel del Tucuman
Reunión:
Simposio; V Simposio Internacional de Bacterias Lacticas SIBAL; 2016
Institución organizadora:
CERELA
Resumen:
Drosophila melanogaster is a reference model organism to decipher complex pathways associated with diseases and metabolic disorders, since more than 75% of its genes have homologues with humans. As well, the functional and structural similarity between Drosophila and vertebrates made the fly a suitable tool to study intestinal infections and diseases, mechanisms of recognition and defense against pathogens, and properties of the natural intestinal microbiota. Lactobacillus plantarum is a gram-positive bacterium colonizing the Drosophila gut with remarkable probiotic qualities among other features. It was suggested that the Lactobacilli ability to colonize intestinal cells is close related to glycogen metabolism. Thus, we were prompted to develop a L. plantarum-Drosophila gnotobiotic model to further study both glycogen metabolism in the bacterium and the impact of this interaction on the physiological and biochemical parameters of Drosophila. Then, in order to achieve the gnotobiotic model we first generated a line of germ free (GF) Drosophila by sequential washing fly embryos with sodium hypochlorite and 70% ethanol. The GF flies were checked by PCR analysis using intestinal tissue as a template for prokaryotic 16s rRNA universal oligonucleotides primers. Afterwards, we proceeded to recolonize the GF flies by adding 50 µl (2x106 CFU/ml) of L. plantarum cultured in MRS broth. To verify the gnotobiotic model, we compared growth curves for the re-colonized flies against GF and wild type flies, in both rich and poor carbohydrates substrates. We observed a clear retardation in GF flies (2 days in the poor carbohydrates substrate) regarding control flies. Remarkable, the gnotobiotic flies recolonized with L. plantarum behaved similarly to the wild type flies. Indeed, this sustain thesustain the idea on the positive effect of L. plantarum in the development of Drosophila melanogaster. To complement this model, we comparatively analyzed biochemical parameters in the fly. Now, we are analyzing the in vitro L. plantarum glycogen metabolism in different carbon sources such as the prebiotics compounds inulin, raffinose and trehalose. Then, lactobacilli grown in these different sources will be analyzed in the gnotobiotic model to shed light regarding the link between prokaryiotic glycogen metabolism, probiotic properties and the impact on the host. We postulate that a better understanding of host?microbe metabolic interactions is important to understand the molecular bases of the metabolic disorders, potentially leading to new therapeutic avenues or the discovery of novel properties for improvement of functional foods