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Biology subjects

Schweizer, M.

Publications and source records attributed to Schweizer, M..

At least 19 recordsLinked to original sources

Pervasive effect of bottom trawling on benthic biodiversity unravelled by metabarcoding across the Tree of Life

Bottom trawling is the most widespread human disturbance on the seafloor, known to affect seabed structure and benthic biota. Yet, its impact on benthic communities remains documented mainly for taxa identifiable through morphology, leaving microscopic benthic communities largely overlooked. The Gulf of Lion continental shelf has been trawled for decades by French and Spanish fleets, to the point that encountering untrawled reference areas became elusive. Here, we examined prokaryote, protist and Animalia benthic communities at eight stations, using eDNA metabarcoding of core sediment samples, with five primer-pairs. Benthic communities were cross-referenced with contrasted trawling impacts, integrated across three periods: over one month, three years or ten years preceding the sampling. The community composition was primarily structured by ecological habitats, yet, trawling pressure showed association with prokaryotes and Animalia diversity. Diversity was higher at sites trawled more intensively in the preceding month, whereas long-term trawling was associated with a significant erosion of Animalia alpha diversity. Unexpectedly, communities did not display consistent vertical structure and the relative contributions of trawling and hydrodynamic reworking remain unresolved due to the absence of untrawled sites. Overall, our results show that trawling effects extend to prokaryotic compartments rarely considered in impact assessments, providing a proof of concept for eDNA-based assessment, and underline a pervasive long-term effect on Animalia communities. This study also highlights the importance of the timescale used to quantify fishing pressure and underscores the complexity of impact assessment when monitoring begins too late to establish a clear baseline, particularly when no reference site remains.

genetics↗

Lysosomal proteome and lipidome analyses of intestinal cells reveal the crucial role of bis(monoacylglycero)phosphate for autophagosome-lysosome fusion

The transport of about 70 enzymes to lysosomes depends on mannose 6-phosphate signals formed by GNPTAB. Editing of Gnptab in an intestinal mouse cell line revealed the loss of multiple lysosomal enzymes associated with the accumulation of sphingomyelins, ceramides, and cholesterol, and reduced levels of bis(monoacylglycero)phosphate (BMP) in lysosomal proteomes and lipidomes. By cross-correlation we identified two subsets of lysosomal lipid-modifying enzymes linked with mixed unsaturated or di-monosaturated BMP. Autophagy-related proteins functioning in early stages of autophagosome formation associated with neutral ceramides, whereas proteins involved in autophagosome-lysosome fusion correlated with negatively charged BMPs. Therefore, the high cholesterol and low BMP level of lysosomes might be causal for impaired autophagic flux in GNPTAB deficient cells. We propose a functional axis of three lysosomal proteins as potential target to improve the autophagic flux in cells with dysfunctional lysosomes and in a newly established intestinal organoid model suitable as novel experimental tool.

cell biology↗

Dissecting the molecular triggers of early and late long-term potentiation

The brain stores information by changing the strength of its synapses, a process that has at least two phases: Late long-term potentiation (L-LTP) is thought to result from the consolidation of early LTP (E-LTP), just as long-term memory requires the prior establishment of short-term memory. Recently, inhibitory avoidance experiments under CaMKII inhibition have challenged this notion, demonstrating long-term fear memory without measurable short-term memory. Here we use optogenetic activation and inhibition of CaMKII during induction of spike-timing-dependent potentiation (tLTP) to dissect the signaling pathways. While CaMKII activation in CA1 neurons was sufficient to induce E-LTP, growth of the postsynaptic density and spine neck expansion, we found that CaMKII-induced LTP does not give rise to L-LTP. Conversely, inhibition of CaMKII during tLTP induction prevented E-LTP, but FOS and L-LTP were still expressed, driven by CaMKK and PKM{zeta}. Thus, both long-term memory and L-LTP form in the absence of CaMKII activation.

neuroscience↗

Propagule and Juvenile-derived Foraminiferal eDNA across intertidal habitats and its implications for accurate sea-level reconstruction

Foraminiferal environmental DNA (eDNA) assemblages have recently emerged as a robust and complementary proxy for relative sea level (RSL) reconstruction. However, unlike traditional morphological methods, eDNA assemblages are influenced by diverse DNA sources, including propagules and juveniles, whose effects on RSL reconstruction remain poorly understood. To assess how foraminiferal eDNA from different life stages vary in taxa composition and impact RSL reconstruction, we analyzed foraminiferal eDNA from bulk, 500-63 m and <63 m size fraction sediments from mangrove and mudflat environments in subtropical Hong Kong. The eDNA assemblages in size-fractioned sediments displayed distinct patterns from those in bulk sediment eDNA across different environments. The propagule and juvenile-derived eDNA <63 m fraction exhibited a similar community structure to bulk eDNA in mudflat environments but diverged in mangrove environments, indicating a greater contribution of propagule and juvenile eDNA to the total eDNA pool in the mudflat environment. We applied Bayesian transfer function modeling to estimate the elevation of samples using different size fractions. eDNA assemblages from the <63 m fraction systematically underpredicted elevation in mangrove environments, while elevations inferred from the 500-63 m fraction and bulk sediment eDNA were accurate. Conversely, all eDNA assemblages in the mudflat-mangrove transitional zone led to the overprediction of RSL. These findings confirm the reliability of bulk sediment eDNA for RSL reconstruction in mangrove environments, while highlighting the need for caution when reconstructing RSL in transitional zones.

ecology↗

Temporal and spatial variability of mudflat and mangrove foraminiferal eDNA assemblages and its implication for sea-level reconstruction

Reconstructing relative sea level (RSL) is essential for understanding coastal evolution and mitigating impacts of climate change. Foraminiferal assemblages are established proxies for past sea levels, but their composition can vary seasonally and spatially, affecting the reliability of morphological reconstructions. Environmental DNA (eDNA) enables high-resolution, non-invasive monitoring of foraminiferal communities and supports high-precision RSL reconstruction. However, the spatiotemporal stability of eDNA assemblages in (sub)tropical intertidal zones--and their influence on RSL reconstruction--remains uncertain. We conducted a two-year eDNA monitoring study at three intertidal stations of varying tidal elevation in Hong Kong, sampling during both dry and wet seasons to assess variability in mangrove and mudflat environments. Mid-mangrove eDNA communities exhibited temporal and spatial stability. In contrast, eDNA assemblages in mudflat and upper-mangrove environments, particularly among monothalamous foraminifera taxa, showed pronounced seasonal shifts primarily driven by environmental changes. Despite this variability in the upper-mangrove, eDNA-based elevation estimates in mangrove consistently aligned with observed elevations (within 95% credible intervals), demonstrating reliability of RSL reconstructions in these environments. However, samples from mudflats, especially during the wet season, exhibited an overprediction bias, reflecting their heightened sensitivity to seasonal and exogenous eDNA inputs. These findings highlight the need to account for seasonal and environmental variability in eDNA-based RSL reconstruction. Stable mangroves are optimal for transfer functions, while transitional/mudflat zones require caution due to higher variability. Our study provides guidance for foraminiferal eDNA application in complex, dynamic coastal settings.

ecology↗

Foraminiferal environmental DNA reveals late Holocene sea-level changes

Reconstructing past relative sea level (RSL) provides critical insight into the mechanisms driving RSL change and informs future projections. Foraminifera are widely used sea-level indicators, but their application is often limited by poor preservation. Here, we demonstrate that foraminiferal environmental DNA (eDNA) and sedimentary ancient DNA (sedaDNA) provide a complementary approach to traditional morphological methods for RSL reconstruction. By analyzing surface sediments and a core from subtropical intertidal environments in the Pearl River Delta, we found that foraminiferal eDNA and sedaDNA assemblages exhibit clear vertical zonation, consistent with morphological results. An eDNA-based transfer function enabled high-resolution RSL reconstructions with decadal temporal and decimeter vertical precision from two periods: 290-1703 CE and 1956-present. Notably, eDNA preservation extended RSL reconstruction far earlier than morphological analyses (1956 CE). The eDNA reconstruction closely matched tide-gauge and geological RSL records, underscoring its potential as a robust tool for reconstructing past RSL and its driving mechanisms.

paleontology↗

Ultrastructural analysis of synapses after induction of spike-timing-dependent plasticity

Repeated sequential activation of connected neurons causes lasting changes in synaptic strength, a process known as spike-timing-dependent plasticity (STDP). Recently, sequential spike patterns have been induced without electrodes, using two spectrally separated channelrhodopsins. However, due to the difficulty of labeling and localizing the few connecting synapses between the stimulated pre and postsynaptic neurons ([~]1-5 per neuron pair), ultrastructural analysis after STDP has not been reported. Here, we optogenetically induce STDP at CA3-CA1 hippocampal synapses and identify stimulated boutons and spines in CA1 using transmission electron microscopy (TEM). Presynaptic CA3 neurons express vesicle-targeted horseradish peroxidase, cre recombinase and cre-dependent ChrimsonR, a red light activatable channelrhodopsin. Postsynaptic neurons express violet light activatable CheRiff and dAPEX2, an enhanced ascorbate peroxidase. In transmission electron microscopy, presynaptic boutons and postsynaptic spines are readily identifiable with well-preserved ultrastructural features. Our labeling strategy allows ultrastructural analysis of optogenetically manipulated neurons and their synapses.

neuroscience↗

Pathological meprin α expression associated with degradation of dermokine drives a psoriasis-like skin phenotype in a genetic mouse model

Keratinocyte proliferation and differentiation is regulated via proteolytic networks. Dysregulation of proteases within these networks can cause hyperproliferative and inflammatory skin disorders. In healthy skin the metalloprotease meprin is localized in the stratum basale. In contrast, in wound healing tissue and psoriatic lesions increased meprin levels are found in the upper epidermal layers. We developed a transgenic mouse model for inducible expression of pathological meprin levels (K5M) to investigate its epidermal degradome and identify molecular links to keratinocyte proliferation and skin inflammation. K5M mice developed a severe skin phenotype characterized by hyperkeratosis, acanthosis and parakeratosis accompanied by increased transepidermal water loss and a strong inflammatory response within six days after induction of meprin overexpression. Histological and molecular analyses showed that increasing meprin expression correlates with meprin activity and keratinocyte hyperproliferation. Proteomics analyses revealed massive changes in proteins associated with keratinocyte differentiation and epidermal barrier integrity already three days after induction. N-terminomics data indicated a dominant chymotryptic activity with elevated proteolytic turnover of proteins associated with the cytoskeleton, cellular stress responses and cell adhesion. By filtering for cleavage sites that match with the specificity of meprin , we identified highly elevated dermokine-derived peptides. Subsequent mass spectrometric analyses validated dermokine as a novel substrate of meprin and identified the cleavage site, which is highly conserved in mammals. Based on the striking similarities with the phenotype reported for dermokine {beta}{gamma}-/- mice, we propose meprin as a central regulator of keratinocyte proliferation and leukocyte recruitment by proteolytic inactivation of dermokine. Hence, pathological meprin activity could be a driver of hyperproliferative, inflammatory skin disorders like psoriasis vulgaris.

molecular biology↗

The Molecular Architecture of Somatic Spines of the Lateral Septum

The lateral septum is a key subcortical structure and has been implicated in kinship memory. Across species, the ability to recognise relatives is conserved and is reflected in stable, lifelong memories. The molecular mechanisms underlying kinship memory remain elusive. Here we investigate the synaptic architecture of septal circuits and focus on somatic spines, an apparent synaptic specialisation of this mainly GABAergic structure. We uncover the molecular organisation of septal somatic spines found in the lateral septum using a combination of unbiased imaging methods and confocal microscopy. We used classical label-free methods such as transmission electron microscopy and Golgi stainings and established new culturing methods for dissociated and organotypic septal slice cultures that were kept in culture over multiple weeks. We describe the presence, morphology, ultrastructure and molecular composition of excitatory somatic spines across multiple developmental stages in various model systems. While smaller than dendritic spines, somatic spines exhibited distinct features, frequently containing secretory organelles such as phagophores and endosomes, but often lack a spine apparatus and ribosomes. Our findings offer insights on the molecular architecture of septal somatic spines and establish a basis for further investigations on the somatic spines of the lateral septum and their role in kinship memory. Significance statementSpine synapses represent stable synaptic connections that allow neurons to chemically communicate with each other. Although known for decades, the unusual spine synapses found directly on neuronal cell bodies in the lateral septum have not been characterised in detail. Using traditional, unbiased approaches such as electron microscopy and Golgi stainings, we replicate the initial findings. Additionally, using fluorescent microscopy of dissociated and organotypic septal cultures, we characterise the development and composition of septal somatic spines. We find that they are excitatory, contain membranous organelles and develop independently of extra-septal input. We thus describe model systems suitable for the investigation of somatospiny neurons of the lateral septum.

neuroscience↗

Extracellular vesicles released from cortical neurons influence spontaneous activity of recipient neurons.

Extracellular vesicles (EVs) are membranous structures that cells release into the extracellular space. EVs carry various molecules such as proteins, lipids, and nucleic acids, and serve as specialized transporters to influence other cells. In the central nervous system, EVs have been linked to many important processes, including intercellular communication, but molecular details of their physiological functions are not fully understood. Our study aimed to investigate how EVs are released by neuronal cells, and how they affect the neuronal activity of other recipient neurons. We show that mature primary cortical neurons release EVs from both their soma and dendrites. EVs released from neurons closely resemble non-neuronal EVs regarding size and marker proteins and proteomic analyses showed that neuronally released EVs contain proteins typically acting in pre- and post-synaptic compartments. Interestingly, our analysis revealed that EVs alter spontaneous activity in target neurons by increasing the amplitude of postsynaptic potentials. In summary, our findings elaborates on the role of EVs in synaptic activity modulation in neurons mediated by glutamate receptors.

neuroscience↗

Optimization and evaluation of complementary degrader discovery assays for application in screening

Targeted protein degradation (TPD) mediated by molecular glues is an innovative pharmaceutical paradigm. By binding to and modulating the surface of an E3-ligase component, molecular glue degraders can facilitate the recruitment of a specific target protein (or vice versa) and, ultimately, invoke target degradation. This mode of action results in specific challenges for the development of rational discovery strategies, and complex hit validation workflows may be required to reliably eliminate compounds that elicit non-specific effects. With the aim to guide screening efforts, we optimized two orthogonal cell-based, target-centric assays for degrader discovery: (1) a time-resolved FRET assay directly quantifying levels of target protein and its degradation (signal inhibition), and (2) an assay coupling TPD to cell growth (signal rescue). To enable a deeper understanding of the individual assays strengths and limitations, we compared their statistical performance as well as respective hit populations by screening a specifically designed collection of about 1000 compounds containing well annotated reference compounds and known frequent hitters. We found that the signal rescue format reliably and specifically captured active target degraders while it efficiently filtered out interfering or frequent hitter compounds. Importantly, this format achieved to retrieve lower potency hits, which might be desirable in order to confidently include as many diverse chemical starting points as possible at the start of a drug discovery project.

pharmacology and toxicology↗

Wildlife trade drives animal-to-human pathogen transmission over 40 years

Wildlife trade affects a quarter of terrestrial vertebrates(1) and creates novel opportunities for cross-species pathogen transmission(2-4). Yet, its precise role in shaping human-wildlife pathogen exchanges remains unclear. Analysing 40 years of global wildlife trade data, we show that traded mammals are twice as likely to share pathogens with humans than non-traded species. Moreover, time spent in trade strongly predicts the number of zoonotic pathogens that wildlife species host: on average, a species shares an additional pathogen with humans for every 9 years of presence in trade. This work highlights the importance of the wildlife trade in driving animal-to-human pathogen transmission, offering key insights into pathogen biogeography in the Anthropocene and the prevention of future pandemics.

ecology↗

Non-canonical function of ADAM10 in presynaptic plasticity

A Disintegrin And Metalloproteinase 10 (ADAM10) plays a pivotal role in shaping neuronal networks by orchestrating the activity of numerous membrane proteins through the shedding of their extracellular domains. Despite its significance in the brain, the specific cellular localization of ADAM10 remains not well understood due to a lack of appropriate tools. Here, using a specific ADAM10 antibody suitable for immunostainings, we discover that ADAM10 is localized to presynapses and especially enriched at presynaptic vesicles of mossy fiber (MF)-CA3 synapses in the hippocampus. These synapses undergo pronounced frequency facilitation of neurotransmitter release, a process that play critical roles in information transfer and neural computation. We demonstrate, that in conditional ADAM10 knockout mice the ability of MF synapses to undergo this type of synaptic plasticity is greatly reduced. The loss of facilitation depends on the cytosolic domain of ADAM10 and association with the calcium sensor synaptotagmin 7 rather than its proteolytic activity. Our findings unveil a new pathway contributing to the regulation of synaptic vesicle exocytosis.

neuroscience↗

Efficient enzyme-free isolation of brain-derived extracellular vesicles

Extracellular vesicles (EVs) have gained significant attention as pathology mediators and potential diagnostic tools for neurodegenerative diseases. However, isolation of brain-derived EVs (BDEVs) from tissue remains challenging, often involving enzymatic digestion steps that may compromise the integrity of EV proteins and overall functionality. Here, we describe that collagenase digestion, commonly used for BDEV isolation, produces undesired protein cleavage of EV-associated proteins in brain tissue homogenates and cell-derived EVs. In order to avoid this effect, we studied the possibility of isolating BDEVs with a reduced amount of collagenase or without any protease. Characterization of the isolated BDEVs revealed their characteristic morphology and size distribution with both approaches. However, we revealed that even minor enzymatic digestion induces artificial proteolytic processing in key BDEV markers, such as Flotillin-1, CD81, and the cellular prion protein (PrPC), whereas avoiding enzymatic treatment completely preserves their integrity. We found no differences in mRNA and protein content between non-enzymatically and enzymatically isolated BDEVs, suggesting that we are purifying the same BDEV populations with both approaches. Intriguingly, the lack of Golgi marker GM130 signal, often referred to as contamination contamination-negative marker in EV preparations, seems to result from enzymatic digestion rather than from its actual absence in BDEV samples. Overall, we show that non-enzymatic isolation of EVs from brain tissue is possible and avoids artificial pruning of proteins while achieving a high BDEV yield and purity. This protocol will help to understand the functions of BDEV in a near-physiological setting, thus opening new research approaches.

neuroscience↗

Astrocytic uptake of posttranslationally modified amyloid-β leads to endolysosomal system disruption and induction of pro-inflammatory signaling

The disruption of astrocytic catabolic processes contributes to the impairment of amyloid-{beta} (A{beta}) clearance, neuroinflammatory signaling, and the loss of synaptic contacts in late-onset Alzheimers disease (AD). While it is known that the posttranslational modifications of A{beta} have significant implications on biophysical properties of the peptides, their consequences for clearance impairment are not well understood. It was previously shown that N-terminally pyroglutamylated A{beta}3(pE)-42, a significant constituent of amyloid plaques, is efficiently taken up by astrocytes, leading to the release of pro-inflammatory cytokine tumor necrosis factor (TNF) and synapse loss. Here we report that A{beta}3(pE)-42, but not A{beta}1-42, gradually accumulates within the astrocytic endolysosomal system, disrupting this catabolic pathway and inducing formation of heteromorphous vacuoles. This accumulation alters lysosomal kinetics and lysosome-dependent calcium signaling, and upregulates lysosomal stress response. These changes correlate with the upregulation of glial fibrillary acidic protein (GFAP) and increased activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NFB). Treatment with a lysosomal protease inhibitor, E64, rescues GFAP upregulation, NFB activation, and synapse loss, indicating that abnormal lysosomal protease activity is upstream of pro-inflammatory signaling and related synapse loss. Collectively, our data suggest that A{beta}3(pE)-42-induced disruption of the astrocytic endolysosomal system leads to cytoplasmic leakage of lysosomal proteases, promoting pro-inflammatory signaling and synapse loss, hallmarks of AD-pathology.

neuroscience↗

TRPML1 activation ameliorates lysosomal phenotypes in CLN3 deficient retinal pigment epithelial cells

Mutations in the lysosomal membrane protein CLN3 cause Juvenile Neuronal Ceroid Lipofuscinosis (JNCL). Activation of the lysosomal ion channel TRPML1 has previously been shown to be beneficial in several neurodegenerative disease models. Here, we tested whether TRPML1 activation rescues disease-associated phenotypes in CLN3-deficient retinal pigment epithelial (ARPE-19 CLN3-KO) cells. ARPE-19 CLN3-KO cells accumulate LAMP1 positive organelles and show lysosomal storage of mitochondrial ATPase subunit C (SubC), globotriaosylceramide (Gb3), and glycerophosphodiesters (GPDs), whereas lysosomal bis(monoacylglycero)phosphate (BMP/LBPA) lipid levels were significantly decreased. Activation of TRPML1 reduced lysosomal storage of Gb3 and SubC but failed to restore BMP levels in CLN3-KO cells. TRPML1-mediated decrease of storage was TFEB-independent, and we identified TRPML1-mediated enhanced lysosomal exocytosis as a likely mechanism for clearing storage including GPDs. Therefore, ARPE-19 CLN3-KO cells represent a human cell model for CLN3 disease showing many of the described core lysosomal deficits, some of which can be improved using TRPML1 agonists.

cell biology↗

Cystatin C loaded in brain-derived extracellular vesicles rescues synapses after ischemic insult in vitro and in vivo

Synaptic loss is an early event in the undersupplied but not yet irreversibly injured penumbra area after an ischemic stroke. Promoting synaptic preservation in this area would likely improve functional neurological recovery. In the present study, we aimed to detect proteins involved in endogenous protection mechanisms of synapses in the penumbra after stroke and to analyse the potential beneficial effect of these candidates for a prospective stroke treatment. For this, we performed Liquid Chromatography coupled to Mass Spectrometry (LC-MS)-based proteomics of synaptosomes isolated from the ipsilateral hemispheres of mice subjected to experimental stroke at different time points (24 h, 4 and 7 days) and compared them to sham-operated mice. Proteomic analyses indicated that among the differentially expressed proteins between the two groups, cystatin C (CysC) was significantly increased at 24 h and 4 days following stroke, before returning to steady-state levels at 7 days, thus indicating a potential transient and intrinsic rescue mechanism attempt of neurons. When CysC was applied to primary neuronal cultures subjected to an in vitro model of ischemic damage, this treatment significantly improved the preservation of synaptic structures. Notably, similar effects were observed when CysC was loaded into brain-derived extracellular vesicles (BDEVs). Finally, when CysC contained in BDEVs was administered intracerebroventricularly to stroked mice, it significantly increased the expression of synaptic markers such as SNAP25, Homer-1, and NCAM in the penumbra area compared to the group supplied with empty BDEVs. Thus, we show that CysC-loaded BDEVs promote synaptic protection after ischemic damage in vitro and in vivo, opening the possibility of a therapeutic use in stroke patients.

neuroscience↗

An annotated chromosome-scale reference genome for Eastern Black-eared Wheatear (Oenanthe melanoleuca)

Pervasive convergent evolution and in part high incidences of hybridization distinguish wheatears (songbirds of the genus Oenanthe) as a versatile system to address questions at the forefront of research on the molecular bases of phenotypic and species diversification. To prepare the genomic resources for this venture, we here generated and annotated a chromosome-scale assembly of the Eastern Black-eared Wheatear (O. melanoleuca). This species is part of the O. hispanica-complex that is characterized by convergent evolution of plumage coloration and high rates of hybridization. The long-read-based male nuclear genome assembly comprises 1.04 Gb in 32 autosomes, the Z chromosome, and the mitogenome. The assembly is highly contiguous (contig N50: 12.6 Mb; scaffold N50: 70 Mb), with 96% of the genome assembled at chromosome level and 96% BUSCO completeness. The nuclear genome was annotated with 18160 protein-coding genes and 30730 mRNAs, and about 10% of the genome consists of repetitive DNA. The annotated chromosome-scale reference genome of Eastern Black-eared Wheatear provides a crucial resource for research into the genomics of adaptation and speciation in an intriguing group of passerines.

genomics↗