Project description:Mass spectrometry imaging is a powerful analytical technique for detecting and determining spatial distributions of molecules within a sample. Typically, mass spectrometry imaging is limited to the analysis of thin tissue sections taken from the middle of a sample. In this work, we present a mass spectrometry imaging method for the detection of compounds produced by bacteria on the outside surface of ant exoskeletons in response to pathogen exposure. Fungus-growing ants have a specialized mutualism with Pseudonocardia, a bacterium that lives on the ants’ exoskeletons and helps protect their fungal garden food source from harmful pathogens. The developed method allows for visualization of bacterial-derived compounds on the ant exoskeleton. This method demonstrates the capability to detect compounds that are specifically localized to the bacterial patch on ant exoskeletons, shows good reproducibility across individual ants, and achieves accurate mass measurements within 5 ppm error when using a high-resolution, accurate-mass mass spectrometer.
2018-07-26 | MTBLS471 | MetaboLights
Project description:Bacterial communities of A.echinatior fungus-growing ants Targeted loci environmental
Project description:Leaf-cutting ants of the genera Acromyrmex and Atta live in mutualistic symbiosis with a basidiomycete fungus (Leucocoprinus gongylophorus), which they cultivate as fungal gardens in underground nest chambers. The ants provide the fungus with a growth substrate consisting of freshly cut leaf fragments. After new leaf fragments are brought into the nest, the ants chew them into smaller pieces and apply droplets of fecal fluid to the leaf pulp before depositing this mixed substrate in the fungus garden and inoculating it with small tufts of mycelium from older parts of the garden. Previous work has shown that the fecal fluid contains a range of digestive enzymes including proteases, amylases, chitinases, cellulases, pectinases, hemicellulases and laccases, and that most of these enzymes are produced by the fungal symbiont in specialized structures called gongylidia that the ants eat. After ingestion, the enzymes apparently pass unharmed through the alimentary channel of the ants and end up in the fecal fluid. Most likely this complex system is an adaptation of the ant-fungus symbiosis to a herbivorous lifestyle, as the ancient ancestors of the ants and the fungus lived as hunter-gatherers and saprotrophs, respectively. The promise of fecal fluid for getting insight into the molecular adaptations that enables the ant-fungus holosymbiont to live as a herbivore, led us to investigate the fecal fluid proteome using LC-MS/MS in order to get a more comprehensive picture of the repertoire of proteins present.
Project description:Leaf-cutter ants use fresh plant material to cultivate an obligate mutualistic fungus Leucoagaricus gongylophorus in specialized fungus gardens to access and transform nutrients from plant biomass that would otherwise be unavailable to the ants. Here, we evaluated the lipidomic differences between the leaves feeding the gardens, gongylidia produced by the fungus to feed the ants, and spatially-resolve regions of the fungus garden at initial to advanced stages of leaf degradation.
Project description:Solid tumors are composed of cancer cells and host immune cells that are distributed in a non-uniform pattern. Growing evidence shows that intratumoral microbes are associated with immune microenvironments in cancer. However, mechanisms that direct the recruitment of microbes to tumors remain poorly understood. Here, we show that intratumoral infiltration of immune cells and microbes are heterogeneous, and the distribution of microbes within tumors are orchestrated by the spatial heterogeneity of intratumoral lymphoid populations. Analysis of human solid tumors revealed that the spatial distribution of immune cells, particularly CD8+ T cells, is markedly heterogeneous. Compared to T cell-poor (“cold”) tumor nests, T cell-rich (“hot”) tumor nests displayed a significantly higher number of myeloid cells, B cells, and plasma cells. We performed laser capture microdissection (LCM) followed by RNA sequencing to identify unique gene signatures that define tumor epithelium and stroma of cold and hot tumor nests. Cold tumor nests expressed genes that promote tumor proliferation and fibrosis, whereas hot tumor stroma and epithelium showed upregulation of immune-related processes, including responses to bacteria, and receptors that mediate mucosal immune responses to microbes, respectively. Consistent with these findings, we detected elevated levels of microbes within hot tumor nests in human pancreatic and lung cancers as well as in mouse models of pancreatic cancer. Intratumoral T cell infiltration plays a causal role in spatial distribution of bacteria in tumor. Our data implicate intratumoral immune heterogeneity in defining microbial spatial distribution and highlight a potential role for crosstalks between microbes, cancer cells, and the host immune system in shaping constituents of the tumor microenvironment (TME).
Project description:This study evaluates the transcriptome of Arabidopsis thaliana seedlings growing in the presence of a diversity of synthetic bacteria communities under different phosphate availability.