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Monitoring Perinatal Gut Microbiota in Mouse Models by Mass Spectrometry Approaches: Parental Genetic Background and Breastfeeding Effects

View Article: PubMed Central - PubMed

ABSTRACT

At birth, contact with external stimuli, such as nutrients derived from food, is necessary to modulate the symbiotic balance between commensal and pathogenic bacteria, protect against bacterial dysbiosis, and initiate the development of the mucosal immune response. Among a variety of different feeding patterns, breastfeeding represents the best modality. In fact, the capacity of breast milk to modulate the composition of infants’ gut microbiota leads to beneficial effects on their health. In this study, we used newborn mice as a model to evaluate the effect of parental genetic background (i.e., IgA-producing mice and IgA-deficient mice) and feeding modulation (i.e., maternal feeding and cross-feeding) on the onset and shaping of gut microbiota after birth. To investigate these topics, we used either a culturomic approach that employed Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MS), or bottom–up Liquid Chromatography, with subsequent MSMS shotgun metaproteomic analysis that compared and assembled results of the two techniques. We found that the microbial community was enriched by lactic acid bacteria when pups were breastfed by wild-type (WT) mothers, while IgA-deficient milk led to an increase in the opportunistic bacterial pathogen (OBP) population. Cross-feeding results suggested that IgA supplementation promoted the exclusion of some OBPs and the temporary appearance of beneficial species in pups fed by WT foster mothers. Our results show that both techniques yield a picture of microbiota from different angles and with varying depths. In particular, our metaproteomic pipeline was found to be a reliable tool in the description of microbiota. Data from these studies are available via ProteomeXchange, with identifier PXD004033.

No MeSH data available.


Diversity index. Box plot of mean values of the Shannon–Wiener index for Balb/c, Rag2ko, Balb/c fed by Rag2ko, and Rag2ko fed by Balb/c mice groups at a genus level at days 3 (A) and 7 (B).
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Figure 4: Diversity index. Box plot of mean values of the Shannon–Wiener index for Balb/c, Rag2ko, Balb/c fed by Rag2ko, and Rag2ko fed by Balb/c mice groups at a genus level at days 3 (A) and 7 (B).

Mentions: The Shannon–Wiener diversity index is a quantitative measure of the alpha diversity in an ecological community such as microbiota. In this study, we used this index to take into account how the microbiota genera were distributed among Balb/c, Rag2ko, Balb/c fed by Rag2ko, and Rag2ko fed by Balb/c mice at 3 and 7 days (Figure 4) of mouse life. At day 3, the value of the diversity index increased in both Balb/c mice and Balb/c mice fed by Rag2ko mice, reflecting an increase of ecology evenness. In Rag2ko mice, and more so in Rag2ko mice fed by Balb/c mice, the indices were low, indicating less diversity in terms of OTUs in these groups (Figure 4A). At 7 days, in Rag2ko mice fed by Balb/c mice, the index was increased, while in the remaining three groups it remained fairly stable (Figure 4B).


Monitoring Perinatal Gut Microbiota in Mouse Models by Mass Spectrometry Approaches: Parental Genetic Background and Breastfeeding Effects
Diversity index. Box plot of mean values of the Shannon–Wiener index for Balb/c, Rag2ko, Balb/c fed by Rag2ko, and Rag2ko fed by Balb/c mice groups at a genus level at days 3 (A) and 7 (B).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5036385&req=5

Figure 4: Diversity index. Box plot of mean values of the Shannon–Wiener index for Balb/c, Rag2ko, Balb/c fed by Rag2ko, and Rag2ko fed by Balb/c mice groups at a genus level at days 3 (A) and 7 (B).
Mentions: The Shannon–Wiener diversity index is a quantitative measure of the alpha diversity in an ecological community such as microbiota. In this study, we used this index to take into account how the microbiota genera were distributed among Balb/c, Rag2ko, Balb/c fed by Rag2ko, and Rag2ko fed by Balb/c mice at 3 and 7 days (Figure 4) of mouse life. At day 3, the value of the diversity index increased in both Balb/c mice and Balb/c mice fed by Rag2ko mice, reflecting an increase of ecology evenness. In Rag2ko mice, and more so in Rag2ko mice fed by Balb/c mice, the indices were low, indicating less diversity in terms of OTUs in these groups (Figure 4A). At 7 days, in Rag2ko mice fed by Balb/c mice, the index was increased, while in the remaining three groups it remained fairly stable (Figure 4B).

View Article: PubMed Central - PubMed

ABSTRACT

At birth, contact with external stimuli, such as nutrients derived from food, is necessary to modulate the symbiotic balance between commensal and pathogenic bacteria, protect against bacterial dysbiosis, and initiate the development of the mucosal immune response. Among a variety of different feeding patterns, breastfeeding represents the best modality. In fact, the capacity of breast milk to modulate the composition of infants’ gut microbiota leads to beneficial effects on their health. In this study, we used newborn mice as a model to evaluate the effect of parental genetic background (i.e., IgA-producing mice and IgA-deficient mice) and feeding modulation (i.e., maternal feeding and cross-feeding) on the onset and shaping of gut microbiota after birth. To investigate these topics, we used either a culturomic approach that employed Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MS), or bottom–up Liquid Chromatography, with subsequent MSMS shotgun metaproteomic analysis that compared and assembled results of the two techniques. We found that the microbial community was enriched by lactic acid bacteria when pups were breastfed by wild-type (WT) mothers, while IgA-deficient milk led to an increase in the opportunistic bacterial pathogen (OBP) population. Cross-feeding results suggested that IgA supplementation promoted the exclusion of some OBPs and the temporary appearance of beneficial species in pups fed by WT foster mothers. Our results show that both techniques yield a picture of microbiota from different angles and with varying depths. In particular, our metaproteomic pipeline was found to be a reliable tool in the description of microbiota. Data from these studies are available via ProteomeXchange, with identifier PXD004033.

No MeSH data available.