Project description:Bathymodiolus mussels inhabiting deep-sea hydrothermal vents harbor bacterial symbionts in their gills, which support the animals’ diet. While the basic mechanisms of energy generation and CO2 fixation that drive these symbioses are largely established, details of molecular interactions between the symbiotic partners and adaptations to their respective habitats remain unknown. In this study, we therefore comparatively examined the genomes and proteomes of two Bathymodiolus hosts and their respective symbionts from different geographical locations. Two mussel species were proteomically compared: i) B. thermophilus mussel containing sulfur-oxidizing symbiont from the east pacific rise. thermophilus and ii) B. azoricus containing thiotrophic and methanotrophic symbionts from the mid-atlantic ridge. Symbionts (for both species) and host components (for B. azoricus) were selectively enriched using a multi-step centrifugation procedure. Enriched host and symbiont fractions along with unenriched gill foot tissue were subject to in-depth semi-quantitative proteomic analyses using the orbitrap and velos mass spectrometers. Proteins were quantified based on their spectral counts using the normalized spectral abundance factor (NSAF) method. We identified common strategies of metabolic interactions that provide mutual nutritional support between host and symbionts, such as the detoxification of ambient sulfide by the Bathymodiolus host, which provides a stable thiosulfate reservoir for the thiotrophic symbionts, and a putative amino acid cycling mechanism that could supply the host with symbiont-derived amino acids. A suite of genes and proteins putatively related to virulence or defense functions was particularly abundant in the B. thermophilus symbiont, compared to its symbiont relatives, and may pose a host species-specific adaptation. Our results reveal both, a high degree of integration between the symbiotic partners, and great potential to adapt to the prevailing environment, which facilitate the holobiont’s survival in its hydrothermal vent habitat.
Project description:Digestive Gland Samples: A manila clam oligo microarray platform (GPL10900) was used to profile gene expression in digestive gland of R. philippinarum. Total RNA was extracted from three (3) independent biological replicates of digestive gland for each sampling site, each consisting of tissue pools of five (5) animals. Statistical analysis with SAM (Significance Analysis of Microarray) identified1,127 probes differentially expressed. Gills Samples: A manila clam oligo microarray platform (GPL10900) was used to profile gene expression in gills of R. philippinarum. Total RNA was extracted from three (3) independent biological replicates of gills for each sampling site, each consisting of tissue pools of five (5) animals. Statistical analysis with SAM (Significance Analysis of Microarray) identified1,127 probes differentially expressed. Digestive Gland Samples: In this study, we analyzed six (6) samples, three (3) pools of digestive gland of Manila clam sampled in Marghera and three(3) pools of digestive gland of Manila clam sampled in Alberoni. Gene expression profiling was performed using the Agilent-019810 Ruditapes philippinarum Oligo Microarray platform (GPL10900) based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. Feature Extraction (FE) 9.5 used XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal. Gills Samples: In this study, we analyzed six (6) samples, three (3) pools of gills of Manila clam sampled in Marghera and three(3) pools of gills of Manila clam sampled in Alberoni. Gene expression profiling was performed using the Agilent-019810 Ruditapes philippinarum Oligo Microarray platform (GPL10900) based on single-colour detection (Cyanine-3 only). Microarrays were scanned with Agilent scanner G2565BA (barcode on the left, DNA on the back surface, scanned through the glass) at a resolution of 5 microns; all slides were scanned twice at two different sensitivity settings (XDRHi 100% and XDRLo 10%); the scanner software created a unique ID for each pair of XDR scans and saved it to both scan image files. Feature Extraction (FE) 9.5 used XDR ID to link the pairs of scans together automatically when extracting data. The signal left after all the FE processing steps have been completed is ProcessedSignal that contains the Multiplicatively Detrended, Background-Subtracted Signal.
Project description:Digestive Gland Samples: A manila clam oligo microarray platform (GPL10900) was used to profile gene expression in digestive gland of R. philippinarum. Total RNA was extracted from three (3) independent biological replicates of digestive gland for each sampling site, each consisting of tissue pools of five (5) animals. Statistical analysis with SAM (Significance Analysis of Microarray) identified1,127 probes differentially expressed. Gills Samples: A manila clam oligo microarray platform (GPL10900) was used to profile gene expression in gills of R. philippinarum. Total RNA was extracted from three (3) independent biological replicates of gills for each sampling site, each consisting of tissue pools of five (5) animals. Statistical analysis with SAM (Significance Analysis of Microarray) identified1,127 probes differentially expressed.
Project description:Colonization of deep-sea hydrothermal vents by invertebrates was made efficient through their adaptation to a symbiotic lifestyle with chemosynthetic bacteria, the primary producers of these ecosystems. Anatomical adaptations such as the establishment of specialized cells or organs have been evidenced in numerous deep-sea invertebrates. However, very few studies detailed global inter-dependencies between host and symbionts in these ecosystems. In this study, we proposed to describe, using a proteo-transcriptomic approach, the effects of symbionts on the deep-sea mussel Bathymodiolus azoricus’ molecular biology. We induced an in situ depletion of symbionts and compared the proteo-transcriptome of the gills of mussels in three conditions: symbiotic mussels (natural population), symbiont-depleted mussels and aposymbiotic mussels
Project description:Host-microbe interactions are virtually bidirectional, benefiting both the host and microbial sides. It is becoming increasingly recognized the influence of the microbe on many aspects of host physiology and diseases, but whether/how the host affects their symbionts is poorly characterized. Here, we reported that the host acts as a critical factor to shape the lifestyle of their symbionts in the Drosophila and bacteria model system. First, we observe that Drosophila larvae play a pivotal role in competing with pathogenic symbionts in the co-existing niche. More specifically, host larvae antagonize symbionts by deconstructing the surface slick, preventing outgrowth and antagonizing the pathogenicity of S. marcescens. Furthermore, Drosophila larvae cause the shift in the transcriptomic profile of S. marcescens, characterized with the upregulated expression of genes related to bacterial proliferation and growth and the downregulated expression of genes related to bacterial pathogenicity. More importantly, advances in bacterial single-cell RNA sequencing provide opportunities to reveal transcriptional variation, including toxic factors, across individual cells and a subpopulation clustering of isogenic bacterial populations. Finally, we found that AMPs from larvae recapitulated the response of S. marcescens to the presence of Drosophila larvae. Altogether, these findings provide an insight into the pivotal roles of the host in influencing the potential pathogens' lifecycle switching from commensalism to pathogenicity, opening the door to a better understanding of the ecological relationships between the host and microbe.
Project description:Host-microbe interactions are virtually bidirectional, benefiting both the host and microbial sides. It is becoming increasingly recognized the influence of the microbe on many aspects of host physiology and diseases, but whether/how the host affects their symbionts is poorly characterized. Here, we reported that the host acts as a critical factor to shape the lifestyle of their symbionts in the Drosophila and bacteria model system. First, we observe that Drosophila larvae play a pivotal role in competing with pathogenic symbionts in the co-existing niche. More specifically, host larvae antagonize symbionts by deconstructing the surface slick, preventing outgrowth and antagonizing the pathogenicity of S. marcescens. Furthermore, Drosophila larvae cause the shift in the transcriptomic profile of S. marcescens, characterized with the upregulated expression of genes related to bacterial proliferation and growth and the downregulated expression of genes related to bacterial pathogenicity. More importantly, advances in bacterial single-cell RNA sequencing provide opportunities to reveal transcriptional variation, including toxic factors, across individual cells and a subpopulation clustering of isogenic bacterial populations. Finally, we found that AMPs from larvae recapitulated the response of S. marcescens to the presence of Drosophila larvae. Altogether, these findings provide an insight into the pivotal roles of the host in influencing the potential pathogens' lifecycle switching from commensalism to pathogenicity, opening the door to a better understanding of the ecological relationships between the host and microbe.
2024-01-17 | GSE232120 | GEO
Project description:Environmental availability of Lucinid symbionts