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The microbiome has been shown to play a role in the homeostasis and homeostatic development of the brain, including neurogenesis and neuromast development [12, 38, 39, 40]. Neuromast development is a differentiation event that takes place in the zebrafish lateral line system and is crucial for the development of sensory systems, including the lateral line, lateral line sensory neurons, and hair cells. Although the mechanisms involved in neuromast development are not completely understood, two key events are the activation of neural progenitor cells to differentiate into sensory neurons and the migration of these progenitors to the lateral line placodes. The Wnt signaling pathway has been implicated in both these events [41, 42, 43, 44]. Our results are consistent with those findings and suggest that the microbiome acts to modulate the development of sensory organs and cell differentiation through Wnt signaling. In addition, the DAVID analysis of the microbiome (Supplemental Fig. 5) revealed the enrichment of genes involved in the TGF-beta signaling pathway (Supplemental Fig. 9). This finding further suggests that the microbiome may affect the development of sensory organs through the TGF-beta signaling pathway, a finding that could be confirmed using additional tools such as specific antibodies and morpholino injections.
Note added in proof: In the course of this study, we became aware of a study in a complementary model system, that used mouse embryos to identify that bacteria can activate Wnt signaling . In addition, we were also able to confirm that the inhibition of Wnt signaling using PKF118-310 blocked neuromast development in the zebrafish. All these studies presented in this manuscript agree with our findings and further support the role of bacteria in activating the Wnt pathway. Moreover, our study provides the first evidence that bacteria activate Wnt signaling in a vertebrate model system.
As bacterial metabolites also alter the immune system, we used DAVID to identify proteins with immune functions that were differentially expressed in response to bacterial metabolites. Of the proteins identified, only one, tbc1d4, was upregulated in the GF treatment.
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One of the drawbacks to this approach was that we needed to recompile every time we made even a small change to the code. This slowed everything else we were doing down and because we couldn't always have someone waiting for the application to finish building. 827ec27edc