Biology subjects

de Jonge, N.

Publications and source records attributed to de Jonge, N..

6 recordsLinked to original sources

bioRxiv2026

Interactions between inbreeding, fitness and the bacterial microbiome in Aedes aegypti mosquitoes

Laboratory and field populations of insects can experience a decline in fitness and loss of genetic diversity due to inbreeding depression and genetic drift, respectively. Matings among related individuals and small population size may also influence insect host microbiomes with consequences for fitness. In the dengue vector mosquito, Aedes aegypti, the bacterial microbiome is largely environmentally determined but recent studies have also revealed host genetic components. We generated a panel of 55 inbred lines from either of two founding outbred populations of Ae. aegypti to test for associations between life history traits, inbreeding, allelic diversity and microbiome composition using ddRADseq and bacterial 16S rRNA gene sequencing on pools of mosquitoes. Effects of inbreeding were diverse with severe composite fitness costs in many lines but minimal costs in others despite similar low levels of genetic diversity. We found no strong relationship between major life history traits across inbred lines, suggesting that any costs due to inbreeding were trait specific. Bacterial microbiome analysis of pooled samples from a subset of populations revealed common microbes across populations, with Elizabethkingia, Aeromonas and Ralstonia being the most abundant. Despite bacterial composition varying widely, there was no clear relationship between microbiome composition and fitness or population origin. However, there were several significant positive correlations between the relative abundance of different microbial taxa across lines. Our results demonstrate diverse impacts of inbreeding on fitness of mosquito populations but with limited impacts on the microbiome.

microbiology↗

bioRxiv2024

Selective enrichment of high-affinity clade II N2O-reducers in a mixed culture

Microorganisms encoding for the N2O reductase (NosZ) are the only known biological sink of the potent greenhouse gas N2O, and are central to global N2O mitigation efforts. Yet, the ecological constraints selecting for different N2O-reducers strains and controlling the assembly of N2O-respiring communities remain largely unknown. Of particular biotechnological interest are clade II NosZ populations, which usually feature high N2O affinities and often lack other denitrification genes. Two planktonic N2O-respiring mixed cultures were enriched under limiting and excess dissolved N2O availability to assess the impact of substrate affinity and N2O cytotoxicity, respectively. Genome-resolved metaproteomics was used to infer the metabolism of the enriched populations. We show that clade II N2O-reducers outcompete clade I affiliates for N2O at sufficiently low sludge dilution rates (0.006 h-1), a scenario previously only theorized based on pure-cultures. Under N2O limitation, all enriched N2O-reducers encoded and expressed only clade II NosZ, while also possessing other denitrification genes. Two Azonexus and Thauera genera affiliates dominated the culture. We explain their coexistence with the genome-inferred metabolic exchange of cobalamin intermediates. Conversely, under excess N2O, clade I and II populations coexisted. Notably, the single dominant N2O-reducer (genus Azonexus) expressed most cobalamin biosynthesis marker genes, likely to contrast the continuous cobalamin inactivation by dissolved cytotoxic N2O concentrations (400 {micro}M). Ultimately, we demonstrate that the solids dilution rate controls the selection among NosZ clades, albeit the conditions selecting for genomes possessing the sole nosZ remain elusive. Additionally, we suggest the significance of N2O-cobalamin interactions in shaping the composition of N2O-respiring microbiomes.

systems biology↗

bioRxiv2020

Transcriptional regulation profiling reveals disrupted lipid metabolism in failing hearts with a pathogenic phospholamban mutation

BackgroundThe R14del mutation in the phospholamban (PLN) gene is associated with various types of cardiomyopathies and increases the risk of developing life-threatening ventricular arrhythmias. In this study, we focused on a homogeneous Dutch founder cohort of genetic cardiomyopathy due to PLN R14del mutation and aimed to study the influence of epigenetic changes from a multi-dimensional perspective. ResultsUsing cardiac tissue of PLN R14del patients and donors, we identified differentially acetylated promoters and enhancers (H3K27ac ChIPseq), annotated enriched transcription factor (TF) binding motifs located in those regions, and identified differentially expressed genes (RNA-seq). In line with the fibrofatty replacement in PLN R14del hearts at the histological level, our integrative analysis detected the downregulation of key TF regulators in fatty acid oxidation (FAO) metabolisms and their downstream target in PLN R14del hearts as compared to controls. We further examined heart tissue using immunofluorescence staining (IF) and to confirm the mitochondrial lipid abnormalities in the PLN R14del hearts. Furthermore, we observed the accumulation and deformation of lipid droplets and a disrupted morphology of mitochondria, the key organelle where FAO takes place, in PLN R14del heart using transmission electron microscopy (TEM). ConclusionUsing multi-omics approaches, we successfully obtained a unique list of chromatin regions and genes, including TF-coding genes, which played important roles in the metabolism-related signalling in PLN R14del hearts.

genomics↗

bioRxiv2020

ORAI1 channels form supra-molecular clusters at rest and agglomerate into distinct plasma membrane areas of oval and strand-like shapes after activation

The Ca2+ selective channel ORAI1 and endoplasmic reticulum (ER)-resident STIM proteins form the core of the channel complex mediating store operated Ca2+ entry (SOCE). Using liquid phase electron microscopy (LPEM) the distribution of ORAI1 proteins was examined at rest and after SOCE-activation at nanoscale resolution. The analysis of over seven hundred thousand of ORAI1 positions showed that already at rest, a majority of the ORAI1 channels formed STIM-independent distinct supra-molecular clusters. Upon SOCE activation and in the presence of STIM proteins, ORAI1 assembled in micron-sized two-dimensional (2D) structures, such as the known punctae at the ER plasma membrane contact zones, but also in divergent structures such as strands, and ring-like shapes. Our results thus question the hypothesis that stochastically migrating single ORAI1 channels are trapped at regions containing activated STIM, and we propose instead that supra-molecular ORAI1 clusters fulfill an amplifying function for creating dense ORAI1 accumulations upon SOCE-activation. STATEMENT OF SIGNIFICANCEORAI1 proteins form channels mediating store operated Ca2+ entry, an important trigger for many cellular functions, especially in the immune system. ORAI1 channels at rest are assumed to be randomly distributed in the plasma membrane, while they accumulate into so-called "punctae" upon activation, where binding by STIM proteins activate the Ca2+ channels. Using liquid phase electron microscopy, we discovered that ORAI1 forms small, elongated clusters indicating the existence of supramolecular assemblies. The role of such supramolecular organization of ORAI1 is possibly an amplifying function for the effective creation of ORAI1 accumulations in punctae, since the binding of only one ORAI1 protein would trap a multiple of channels.

biophysics↗

bioRxiv2020

Correlative Fluorescence and Electron Microscopy of Whole Breast Cancer Cells Reveals Differential Distribution of ErbB2 Dependent on Underlying Actin

Epidermal growth factor receptor 2 (ErbB2) is found overexpressed in several cancers, such as gastric, and breast cancer, and is, therefore, an important therapeutic target. ErbB2 plays a central role in cancer cell invasiveness, and is associated with cytoskeletal reorganization. In order to study the spatial correlation of single ErbB2 proteins and actin filaments, we applied correlative fluorescence microscopy, and scanning transmission electron microscopy (STEM) to image specifically labeled SKBR3 breast cancer cells. The breast cancer cells were grown on microchips, transformed to express an actin-green fluorescent protein (GFP) fusion protein, and labeled with quantum dot (QD) nanoparticles attached to specific anti-ErbB2 Affibodies. Fluorescence microscopy was performed to identify cellular regions with spatially correlated actin and ErbB2 expression. For STEM of the intact plasma membrane of whole cells, the cells were fixed and covered with graphene. Spatial distribution patterns of ErbB2 in the actin rich ruffled membrane regions were examined, and compared to adjacent actin-low regions of the same cell, revealing an association of putative signaling active ErbB2 homodimers with actin-rich regions. ErbB2 homodimers were found absent from actin-low membrane regions, as well as after treatment of cells with Cytochalasin D, which breaks up larger actin filaments. In both latter data sets, a significant inter-label distance of 36 nm was identified, possibly indicating an indirect attachment to helical actin filaments via the formation of heterodimers of ErbB2 with epidermal growth factor receptor (EGFR). The possible attachment to actin filaments was further explored by identifying linear QD-chains in actin-rich regions, which also showed an inter-label distance of 36 nm.

biophysics↗

bioRxiv2018

Biomass segregation between biofilm and flocs improves the control of nitrite-oxidizing bacteria in mainstream partial nitritation and anammox processes

The control of nitrite-oxidizing bacteria (NOB) challenges the implementation of partial nitritation and anammox (PN/A) processes under mainstream conditions. The aim of the present study was to understand how operating conditions impact microbial competition and the control of NOB in hybrid PN/A systems, where biofilm and flocs coexist. A hybrid PN/A moving-bed biofilm reactor (MBBR; also referred to as integrated fixed film activated sludge or IFAS) was operated at 15 {degrees}C on aerobically pre-treated municipal wastewater (23 mgNH4-N{middle dot}L-1). Ammonium-oxidizing bacteria (AOB) and NOB were enriched primarily in the flocs, and anammox bacteria (AMX) in the biofilm. After decreasing the dissolved oxygen concentration (DO) from 1.2 to 0.17 mgO2{middle dot}L-1 - with all other operating conditions unchanged - washout of NOB from the flocs was observed. The activity of the minor NOB fraction remaining in the biofilm was suppressed at low DO. As a result, low effluent NO3- concentrations (0.5 mgN{middle dot}L-1) were consistently achieved at aerobic nitrogen removal rates (80 mgN{middle dot}L-1{middle dot}d-1) comparable to those of conventional treatment plants. A simple dynamic mathematical model, assuming perfect biomass segregation with AOB and NOB in the flocs and AMX in the biofilm, was able to qualitatively reproduce the selective washout of NOB from the flocs in response to the decrease in DO-setpoint. Similarly, numerical simulations indicated that flocs removal is an effective operational strategy to achieve the selective washout of NOB. The direct competition for NO2- between NOB and AMX - the latter retained in the biofilm and acting as a \"NO2-sink\" - was identified by the model as key mechanism leading to a difference in the actual growth rates of AOB and NOB (i.e., NOB < AOB in flocs) and allowing for the selective NOB washout. Experimental results and model predictions demonstrate the increased operational flexibility, in terms of variables that can be easily controlled by operators, offered by hybrid systems as compared to solely biofilm systems for the control of NOB in mainstream PN/A applications.\n\nHighlightsO_LIHybrid PN/A systems provide increased operational flexibility for NOB control\nC_LIO_LIAOB and NOB enrich primarily in the flocs, and AMX in the biofilm (\"NO2-sink\")\nC_LIO_LIAMX use NO2- allowing to differentiate AOB and NOB growth rates\nC_LIO_LIA decrease in DO or an increase in floc removal leads to selective NOB washout from flocs\nC_LIO_LIThe activity of the minor NOB fraction in the biofilm is suppressed at limiting DO\nC_LI\n\n\n\nO_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=63 SRC=\"FIGDIR/small/480780v2_ufig1.gif\" ALT=\"Figure 1\">\nView larger version (21K):\norg.highwire.dtl.DTLVardef@cad5deorg.highwire.dtl.DTLVardef@9efef3org.highwire.dtl.DTLVardef@7409feorg.highwire.dtl.DTLVardef@3c1043_HPS_FORMAT_FIGEXP M_FIG C_FIG

systems biology↗