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Gross, J.

Publications and source records attributed to Gross, J..

At least 19 recordsLinked to original sources

Targeting proteasome composition reprograms tumor-intrinsic inflammation through selective degradation of STAT1

Proteasome regulatory particles determine cellular protein degradation capacities. Yet specific ubiquitin-independent degradation targets of individual proteasome regulatory particles, and physiological roles of such pathways, remain largely elusive. Here, through a chemical biology-driven approach, we identify a covalent small molecule that selectively targets a conformationally dynamic pocket in the non-catalytic proteasome regulator PSME4 and modulates tumor cell proteasome targets. Using this chemical probe together with orthogonal genetic perturbations, we show that blocking or reducing PSME4 stabilizes STAT1, restores interferon-associated transcription, enhances antigen presentation, and disrupts IL-6/STAT3 signaling, collectively reprogramming tumor inflammation and suppressing tumor growth in vivo. These findings define a PSME4-STAT1-STAT3 axis that couples proteasomal degradation to antigen presentation and cytokine signaling and provide a framework for pharmacologically targeting non-catalytic proteasome regulators to reprogram tumor immunity. More broadly, our findings reveal that proteasome composition can actively govern the balance between immunostimulatory and immunosuppressive signaling programs.

cancer biology↗

Multi-Methodological Characterization of Sleep Deprivation: From Standard EEG Power Spectra to Aperiodic Dynamics in Humans and Mice.

Sleep deprivation is a potent, rapid-acting therapeutic intervention for major depressive disorder, yet its underlying neural mechanisms remain poorly understood, hindering the development of predictive biomarkers. Here, we systematically characterize the electro-physiological signatures of prolonged wakefulness using a multi-methodological approach across three independent datasets in humans and mice. By integrating standard power spectral density analysis with aperiodic component fitting (SpecParam) and highly compar-ative time-series analysis (HCTSA), we identified robust cross-species biomarkers of sleep pressure. Machine learning models revealed that theta power is the most consistent feature for differentiating control and sleep deprivation states, achieving up to 90% classification accuracy. Sleep deprivation significantly increased the spectral offset - suggesting global cortical hyperexcitation - while simultaneously steepening the spectral slope. We interpret this simultaneous shift as a state uncoordinated state of hyperexcited and inefficient neu-ral processing. These findings establish reproducible EEG markers of sleep deprivation that transcend species. Given the clinical utility of wake therapy, we propose that prefrontal the-ta power and spectral offset/slope may serve as mechanism-based predictors of therapeu-tic response. Our results provide a framework for the clinical validation of these biomarkers, potentially enabling personalized chronotherapeutic interventions for psychiatric disorders.

neuroscience↗

EasyCAPS: A web tool for restriction-based genotyping and rational CRISPR-Cas9 donor design

Tracking Single Nucleotide Polymorphisms (SNPs) following CRISPR-Cas9 genome editing is a critical yet often labor-intensive step in modern genetic research. Although Sanger sequencing is the conventional method for definitive confirmation, it typically requires substantial time to generate results. In contrast, PCR-based restriction methods like CAPS (Cleaved Amplified Polymorphic Sequence) and dCAPS (derived CAPS) offer rapid and cost-effective alternatives. However, existing dCAPS primer design tools suffer from significant limitations and were largely developed for tracking polymorphisms in plant genomes. Concurrently, CRISPR-Cas9 gene editing requires strategies to prevent the re-cleavage of the edited allele, typically involving the modification of the Protospacer Adjacent Motif (PAM). To address these challenges, we developed EasyCAPS, a web-based tool that integrates dCAPS primer design with advanced functionalities for CRISPR experiments. EasyCAPS overcomes the shortcomings of previous software by enabling restriction enzyme pre-selection and optimizing designs for complex DNA sequences. Its key innovation is the "Hiding PAM" feature, which designs synonymous mutations to mask the Cas9 recognition site while accounting for codon usage bias, thereby facilitating one-step allelic exchange. The utility of the tool was demonstrated through practical applications targeting the HTA1, PHO84, and CAT5 genes, significantly accelerating both genotyping and gene editing processes. We conclude that EasyCAPS is an accessible solution that effectively streamlines molecular biology workflows.

genetics↗

Brain anatomy and molecular signaling predict neurophysiological dynamics across the lifespan

Neural activity emerges from interactions between local cellular architecture, neuromodulatory systems, and large-scale cortical networks. Yet it remains unclear how this multiscale biological context constrains electrophysiological dynamics in humans and how this changes across the lifespan. We combined resting-state magnetoencephalography (MEG) from 350 adults (18-88 years) with cortical maps reflecting cytoarchitecture, myelination, metabolism, gene expression, and neurotransmitter receptors in a multivariate prediction framework. Specific markers explained most regional variance in MEG power spectra and temporal autocorrelation, similarly for both measures, revealing frequency- and timescale-specific signatures that followed canonical spectral boundaries. Age-related MEG patterns spatially aligned with markers of neuroinflammation, monoaminergic-cholinergic signaling, cortical development and myelination, and cerebrovascular organization. This work identifies key components of an anatomical and molecular scaffold and their relative importance for neural activity across the lifespan, informing future experimental perturbations and generative models.

neuroscience↗

Backwards compatibility to classical experiments grounds beta responses to naturalistic speech in temporal acoustic forecasting

Current neuroscience is shifting from simple controlled paradigms towards rich and ecologically valid naturalistic stimuli. Correspondingly, insights from historic "impoverished" artificial paradigms are considered to be seriously challenged by generalisation to modern naturalistic contexts. Here, we argue that the reverse, backwards compatibility, is an under-appreciated and easily accessible benchmark and show that it disambiguates model comparison beyond naturalistic stimuli. We analyse magnetoencephalography (MEG) beta power responses to an audiobook stimulus using canonical correlation analysis (CCA) and replicate reported links between beta power and syntactic parsing. However, a simple stimulus-computable acoustic model predicts the same variance, suggesting a domain-general rather than linguistic function of beta responses. We therefore test the backwards compatibility of speech-trained models to classic rhythmic tones. While generalisation initially fails, reducing hidden and unnecessary degrees of freedom of the models phase responses allows successful generalisation. Crucially, this greatly improves model adjudication: Several models that perform indistinguishably on speech differ in how well they predict responses to simple tones. In this comparison, a simple forecasting deep neural network (DNN) outperforms acoustics by internalising a "slow-decay" prior as a structural mirror of sluggish speech dynamics. This grounds beta responses in canonical temporal forecasting, bridging modern naturalism to established auditory psychophysics.

neuroscience↗

The DES-p.A120P mutation associated with biventricular arrhythmogenic cardiomyopathy has a dominant-negative effect on desmin filament assembly

Desmin is a muscle-specific intermediate filament protein, which connects different cell organelles and is highly relevant for the structural integrity of cardiomyocytes. Mutations in the DES gene, cause different cardiomyopathies including arrhythmogenic cardiomyopathy. In this study, we functionally investigate the novel genetic variant DES-p.A120P using cell transfection experiments including cardiomyocytes derived from induced pluripotent stem cells in combination with confocal microscopy. These experiments reveal that the filament assembly of desmin-p.A120P is disturbed - even when co-expressed with wild-type desmin. In conclusion, the functional characterization of desmin-p.A120P supports the classification as a pathogenic variant associated with arrhythmogenic cardiomyopathy.

genetics↗

Lethal effects of the wild potato Solanum bulbocastanum on the planthopper Pentastiridius leporinus, a vector of bacterial pathogens in potato

The planthopper Pentastiridius leporinus [Hemiptera: Cixiidae] is a vector of the two plant pathogenic bacteria Candidatus Arsenophonus phytopathogenicus and Candidatus Phytoplasma solani in potato. An infection leads to symptoms such as rubbery and non-processable tubers and consequently to substantial decreases in yield and tuber quality as well as to abandonment of seed potato production in affected areas. Up to date, resistant potato varieties are not yet known, and the tool box of integrated pest management to prevent the spread of P. leporinus as causal vector is quite limited. Mortality experiments designed as no-choice trials on potato cultivar Gala (Solanum tuberosum) and on its wild relative S. bulbocastanum revealed significant effects of the latter on the survival rate of the vector. LC/MS analysis of the insects and their intestines showed that acyl solamines were retrieved only in individuals from the S. bulbocastanum plants. Furthermore, choice-tests were carried out to study the vector behavior. A preference towards S. bulbocastanum vs. S. tuberosum Gala was observed despite its lethal effects. Volatile organic compounds of the two different species were collected and analyzed by GC/MS. The two profiles differed in 39 of 80 compounds. The wild potato species S. bulbocastanum and its compounds are discussed as potential starting points for further research on sustainable management of P. leporinus. Key MessageO_LIWe tested a wild potato against a planthopper which vectors pathogenic bacteria to potato C_LIO_LIWild potato led to higher mortality of the planthopper but was preferred against cultivated potato C_LIO_LIUptake assays suggest that the phloem sucking planthopper is taking up xylem sap as well C_LIO_LIAcyl solamines were only found in guts from planthoppers sucking on the wild potato C_LIO_LIOur results present a starting point for new approaches for sustainable vector control C_LI

ecology↗

Oviposition behavior, nymphal development, and below-ground olfactory preference of Pentastiridius leporinus on twelve Beta vulgaris genotypes

The planthopper Pentastiridius leporinus (Hemiptera: Cixiidae) is the primary vector of two phloem-restricted prokaryotes, which can cause the disease syndrome de basses richesses (SBR) in sugar beet (Beta vulgaris L.). Heavy infestations of this insect have contributed to significant yield losses in Central Europe, but little is known about how different B. vulgaris genotypes influence vector behavior and performance. Understanding such interactions is critical for both pest management and the development of sugar beet cultivars that are less favorable to the vector. We investigated oviposition decisions, nymphal development, and belowground chemotaxis of P. leporinus using twelve B. vulgaris genotypes: eight sugar beet genotypes with contrasting SBR tolerance, two Swiss chard, and two beetroot varieties. In no-choice assays, females oviposited at similar rates on all tested genotypes, indicating broad host suitability. However, in three-choice tests, significant differences emerged, with several genotypes receiving more egg batches than the reference variety Vasco. Despite adult host preferences, nymphal survival, growth, and instar development did not differ among genotypes. In contrast, belowground olfactometer assays revealed that nymphs exhibited clear chemotactic responses: several genotypes attracted significantly more individuals than others. This represents the first documentation of chemotactic host location in P. leporinus nymphs. Our findings reveal a mismatch between adult oviposition preference and nymphal performance, a pattern not uncommon in other generalist herbivores. Both adult and nymphal behaviors were nevertheless influenced by genotype-specific traits, likely linked to plant chemistry. Identifying these cues could inform breeding strategies aimed at reducing vector attraction, thereby complementing pathogen-targeted approaches and supporting integrated management of SBR in sugar beet.

ecology↗

QTL mapping, breeding, and debugging Saccharomyces cerevisiae strains through Reiterated Mass Selection and backcrosSing (ReMaSSing)

BackgroundProducing second-generation ethanol from lignocellulosic hydrolysates (LCHs) poses significant challenges for Saccharomyces cerevisiae due to the presence of fermentation inhibitors. Quantitative trait loci (QTL) mapping of stress-tolerant S. cerevisiae strains is important for identifying adaptive alleles that can enhance yeast fermentation of LCHs. However, the QTL mapping process is labor-intensive, requiring the screening of numerous recombinants and repeated crossings to improve mapping resolution. ResultsWe developed Reiterated Mass Selection and backcrosSing (ReMaSSing) to facilitate the identification of adaptive alleles through QTL mapping and to enhance LCH tolerance in yeast strains. ReMaSSing was applied to populations obtained by crossing the stress-resistant yeast PE-2_H4 with the laboratory strain S288C. Using alternative protocols, we selected haploid or diploid populations with dominant markers, enriching millions of segregants carrying adaptive alleles by propagating them in standard or LCH-supplemented media. The enriched pools were then bulk backcrossed with S288C, and germination of millions of spores generated new recombinant populations for subsequent selection cycles. After five rounds of ReMaSSing, whole-genome sequencing and QTL mapping identified key alleles associated with LCH tolerance, linked to VPS70, CAT5, GCY1, UBP2, MKT1/SAL1, HAP1, and PHO84, which influence growth and mitochondrial function in S288C. Mutations in IRA1 and HTA1, unique to our S288C strain, were also mapped, highlighting ReMaSSings ability to detect and correct deleterious alleles ("bugs"). Allele swapping and competition assays confirmed that the identified QTL improved LCH tolerance and growth, with strains combining adaptive alleles performing over 20% better than the parental S288C. Finally, applying ReMaSSing to breed an LCH-tolerant yeast with a xylose-consuming strain produced recombinants with improved fermentation of xylose-enriched LCH. ConclusionReMaSSing offers a practical protocol for generating QTL mapping populations to identify adaptive alleles in tolerant strains and correct genetic defects in inferior ones. Notably, recombinant populations and clones derived from ReMaSSing outperformed both parental strains in LCH tolerance and growth. Furthermore, we applied ReMaSSing to breed strains with enhanced LCH tolerance, efficient xylose catabolism, and robust ethanol production. Together, these results demonstrate that ReMaSSing is a powerful tool for engineering industrial yeast strains that integrate desirable traits from multiple parental backgrounds.

genomics↗

Respiration as a dynamic modulator of sensory sampling

Respiration dynamically modulates sensory perception by orchestrating transient states of the brain and the body. Using simultaneous recordings of high-density magneto-encephalography (MEG), respiration, and pupillometry, we show that human perceptual sensitivity to near-threshold visual stimuli was enhanced during inspiration, coinciding with respiration-modulated increases in arousal neuromodulation and cortical excitability. Participants adapted their breathing patterns to align with predictable stimulus onset, and this adaptive respiratory control correlated with improved performance. We further reveal that respiration-modulated changes in alpha and beta oscillations reflect distinct shifts in sensory and motor excitability, respectively. Crucially, respiration-resolved multivariate Granger causality analyses demonstrate that the breathing rhythm systematically shapes directed information flow within a widespread interoceptive network. This respiration-brain coupling was flexibly adjusted based on stimulus predictability, highlighting a novel mechanism for active sensing which integrates internal bodily rhythms with external sensory demands to optimize perception.

neuroscience↗

Structural and mechanistic analysis of covalent ligands targeting the RNA-binding protein NONO

RNA-binding proteins (RBPs) play important roles in mRNA transcription, processing, and translation. Chemical tools are lacking for RBPs, which has hindered efforts to perturb and understand RBP function in cells. We recently reported a chloroacetamide compound (R)-SKBG-1 that covalently binds the RBP NONO and stabilizes its interactions with mRNAs, leading to transcriptional remodeling and the suppression of cancer cell growth. Here, we report the crystal structure of an (R)-SKBG-1:NONO complex, which confirms covalent modification of cysteine-145 at a pocket proximal to the RNA-binding interface of the protein. We show that this pocket can also be targeted by a lower reactivity chlorofluoroacetamide analog (R, R)-GL-373, which retains the pharmacological properties of (R)-SKBG-1, including blockade of estrogen receptor expression in breast cancer cells, while displaying much greater proteome-wide selectivity. Our findings thus show that NONO can be targeted by covalent ligands with high specificity to pharmacologically suppress pro-tumorigenic gene products in cancer cells. HighlightsO_LICo-crystal structure reveals how covalent ligands stereoselectively bind NONO C_LIO_LIChlorofluoroacetamides (CFAs) bind NONO with high proteomic selectivity C_LIO_LICFA ligands remodel cancer cell transcriptomes in a NONO-dependent manner C_LIO_LICFA ligands suppress cancer cell growth in a NONO-dependent manner C_LI

biochemistry↗

Hidden Bias: GPCR Polymorphisms and the Genomic Landscape of β- Arrestin Signaling

Clinically, no human diseases are currently diagnosed as being directly driven by {beta}- arrestins. Nonetheless, there is growing interest in developing {beta}-arrestin-biased drugs, due to the key regulatory role of {beta}-arrestins in modulating the cellular signaling of most G protein-coupled receptors (GPCRs). In cell-based studies of rhodopsin-family GPCRs, mutations in a conserved proline-hydrophobic (ProH) motif within the second intracellular loop (ICL2) have been shown to alter {beta}-arrestin signaling bias. The clinical relevance of such mutations in humans is unknown. However, if naturally occurring single nucleotide polymorphisms (SNPs) affecting this ProH motif exist across the GPCR family, then cross-referencing these genetic variants with large-scale population sequencing and epidemiologic data could reveal potential roles for {beta}-arrestin signaling in human health. In this report, we identify SNPs in human GPCRs that correspond to ProH substitutions, and we show that, in the neurotensin receptor NTSR1, these variants can indeed shift signaling bias. We also estimate a lower-bound population frequency for such SNPs, suggesting that although rare for any given receptor, their cumulative prevalence across the GPCR superfamily may be large enough to impact phenotypic variation. Together with emerging data from biased ligands, our findings support the idea that genetic variations in {beta}-arrestin signaling could represent a meaningful source of therapeutic relevance. Significance statementMutations altering G protein-coupled receptor (GPCR) signaling can affect health. We have identified a specific amino acid determinant in intracellular loop (ICL2) of human rhodopsin family GPCRs that influences {beta}-arrestin signaling. Although the medical consequences of this determinant remain unclear, we show that single nucleotide polymorphisms (SNPs) affecting the determinant routinely occur. By studying neurotensin receptor NTSR1, we confirm that the SNP alters {beta}-arrestin signaling bias. While individually rare per receptor, these SNPs may collectively contribute to {beta}-arrestin-related phenotypic changes in the human population. Our findings, combined with research on biased drugs, support the idea that {beta}-arrestin signaling might serve as a useful therapeutic target, opening new possibilities for precision medicine.

biochemistry↗

Complexoform-restricted covalent TRMT112 ligands that allosterically agonize METTL5

Adaptors serve as hubs to regulate diverse protein complexes in cells. This multitude of functions can complicate the study of adaptors, as their genetic disruption may simultaneously impair the activities of several compositionally distinct complexes (or adaptor complexoforms). Here we describe the chemical proteomic discovery of bicyclopyrrolidine acrylamide stereoprobes that react with cysteine-100 (C100) of the methyltransferase (MT) adaptor TRMT112 in human cells. Curiously, the stereoprobes showed negligible reactivity with uncomplexed recombinant TRMT112, and we found that this interaction was restored excluively in the presence of METTL5, but not other MTs. A co-crystal structure revealed stereoprobe binding to a composite pocket proximal to C100 of TRMT112 that is templated by METTL5 and absent in other TRMT112:MT complexes. Structural rearrangements promoted by stereoprobe binding in turn lead to allosteric agonism of METTL5, thus revealing how covalent ligands targeting a pleiotropic adaptor can confer partner-specific functional effects through reactivity with a single complexoform.

biochemistry↗

Zebrafish optic nerve injury results in systemic retinal ganglion cell dedifferentiation

Retinal ganglion cells (RGCs) are the sole projection neurons connecting the retina to the brain and therefore play a critical role in vision. Death of RGCs during glaucoma, optic neuropathies and after ocular trauma results in irreversible loss of vision as RGCs do not regenerate in the human eye. Moreover, there are no FDA approved therapies that prevent RGC death and/or promote RGC survival in the diseased or injured eye. There is a critical need to better understand the molecular underpinnings of neuroprotection to develop effective therapeutic approaches to preserve damaged RGCs. Unlike in mammals, RGCs in zebrafish are resilient to optic nerve injury, even after complete transection of the optic nerve. Here, we leveraged this unique model and utilized single-cell RNA sequencing to characterize RGC responses to injury and identify putative neuroprotective and regenerative pathways. RGCs are heterogeneous and studies in mice have shown that there is differential resiliency across RGC subtypes. Our results demonstrated that all RGC subtypes are resilient to injury in zebrafish. Quantifying changes in gene expression revealed the upregulation of progenitor and regenerative markers in all RGC subtypes after injury as well as distinct early and late phases to the injury response. This shift in gene expression causes injury-responsive RGCs to resemble RGC subtype 3, a low frequency population of endogenous immature RGCs that are normally maintained in the wild-type, uninjured adult retina. A similar but restricted transcriptomic injury response in RGCs of the uninjured contralateral eye was also detected, highlighting a systemic RGC response to unilateral optic nerve injury. Taken together, these results demonstrate that zebrafish RGCs dedifferentiate in response to injury, and this may be a novel mechanism mediating their unique cell survival and regenerative capabilities. Author SummaryRetinal ganglion cells (RGCs) connect the eye to the brain and are essential for vision. Their death in conditions like glaucoma, affecting over 70 million people worldwide, leads to permanent blindness, with no FDA-approved treatments to prevent it. Unlike mammals, zebrafish RGCs are resilient to optic nerve injury. In this study, we used next-generation sequencing technologies to characterize the RGC response to optic nerve injury at the single-cell level. We discovered that all zebrafish RGCs survive damage by temporarily shifting into a less mature state, resembling a rare population of immature RGCs found in uninjured animals. We identified many genes whose expression changes early or late in the injury response as well as a similar but restricted transcriptomic injury response in the uninjured contralateral RGCs, highlighting the systemic RGC response to optic nerve injury. This work is significant because our detailed characterization of RGC responses to optic nerve injury identifies dedifferentiation as an injury response, possibly important for cell survival and axon regrowth. The genes and pathways we identify are potential therapeutic targets to enable RGC survival in the injured or diseased human eye.

neuroscience↗

Engineering Adaptive Alleles for Escherichia coli Growth on Sucrose Using the EasyGuide CRISPR System

Adaptive Laboratory Evolution (ALE) is a powerful approach for mining genetic data to engineer industrial microorganisms. This evolution-informed design requires robust genetic tools to incorporate the discovered alleles into target strains. Here, we introduce the EasyGuide CRISPR, a five-plasmid platform that exploits E. colis natural recombination system to assemble gRNA plasmids from overlapping PCR fragments. The production of gRNAs and donor DNA is further facilitated by using recombination cassettes generated through PCR with 40 to 60-mer oligos. With the new CRISPR toolkit, we constructed 22 gene edits in E. coli DH5, most of which corresponded to alleles mapped in E. coli DH5 and E2348/69 ALE populations selected for sucrose propagation. For DH5 ALE, sucrose consumption was supported by the cscBKA operon expression from a high-copy plasmid. During ALE, plasmid integration into the chromosome, or its copy number reduction due to the pcnB deletion, conferred a 30-35% fitness gain, as demonstrated by CRISPR-engineered strains. A [~]5% advantage was also associated with a [~]40.4 kb deletion involving fli operons for flagella assembly. In E2348/69 ALE, inactivation of the hfl system suggested selection pressures for maintaining {lambda}-prophage dormancy (lysogeny). We further enhanced our CRISPR toolkit using yeast for in vivo assembly of donors and expression cassettes, enabling the establishment of polyhydroxybutyrate synthesis from sucrose. Overall, our study highlights the importance of combining ALE with streamlined CRISPR-mediated allele editing to advance microbial production using cost-effective carbon sources.

genetics↗

tet2 and tet3 regulate cell fate specification and differentiation events during retinal development

AbstractTet enzymes are epigenetic modifiers that impact gene expression via 5mC to 5hmC oxidation. Previous work demonstrated the requirement for Tet and 5hmC during zebrafish retinogenesis. tet2-/-;tet3-/-mutants possessed defects in the formation of differentiated retinal neurons, but the mechanisms underlying these defects are unknown. Here, we leveraged scRNAseq technologies to better understand cell type-specific deficits and molecular signatures underlying the tet2-/-;tet3-/- retinal phenotype. Our results identified defects in the tet2-/-;tet3-/- retinae that included delayed specification of several retinal cell types, reduced maturity across late-stage cones, expansions of immature subpopulations of horizontal and bipolar cells, and altered biases of bipolar cell subtype fates at late differentiation stages. Together, these data highlight the critical role that tet2 and tet3 play as regulators of cell fate specification and terminal differentiation events during retinal development.

developmental biology↗

Personalized tDCS targeting visual motion area V5 modulates smooth pursuit initiation

Transcranial direct current stimulation (tDCS) for the modulation of ongoing eye movements provides an ideal model for investigating sensorimotor integration. Within neural networks subserving smooth pursuit eye movements, visual motion area V5 is a core hub to integrate visual motion information with oculomotor control. Here, we applied personalized tDCS explicitly targeting individual V5 in healthy human participants using algorithmic optimization informed by functional magnetic resonance imaging and combined electro- and magnetencephalography. We hypothesized subtle modulation of sensorimotor integration during pursuit and assessed the gain by personalized tDCS targeting V5, compared to personalized tDCS targeting the frontal eye field, as well as conventional normative tDCS over V5. Indeed, pursuit initiation was specifically delayed during personalized cathodal tDCS targeting right V5 indicating the involvement of distinct functional subregions of V5 in the initial sensorimotor integration of visual motion information with pursuit eye movements, but not the maintenance of ongoing pursuit. The results were well-controlled by anodal and sham tDCS, different pursuit tasks, finite-element simulations of individual electric fields, and by two additional control experiments, one that applied personalized tDCS targeting frontal eye field and another that applied normative tDCS over V5. Importantly, in contrast to personalized tDCS targeting FEF and normative tDCS over V5, personalized tDCS targeting V5 effectively modulated pursuit by adapting electric fields to individual anatomical and functional V5 properties. Our results provide evidence for the specific involvement of area V5 in sensorimotor integration during pursuit initiation and the ability of (targeted) tDCS to specifically introduce subtle modulation of the brain network underlying smooth pursuit eye movements. Further, our results indicate the potential of personalized tDCS to alter behavior as the main aspect of interest in human neuromodulation.

neuroscience↗

Development of a protein synthesis network at the sarco/endoplasmic reticulum in adult cardiac myocytes

IntroductionThe endoplasmic reticulum (ER) is the site of synthesis and folding of membrane and secretory proteins, which constitute a large fraction of the total protein output of a mammalian cell. Striated muscle cells contain a specialized membrane system known as the sarcoplasmic reticulum (SR) that controls calcium homeoastasis and contraction, however the biochemical and physiological relationship between the ER and SR and how both compartments participate in protein synthesis remains incompletely understood. MethodsProtein quantification and imaging of selected marker proteins for ER and SR functions was performed to characterize the relationship of the ER and SR and its individual involvement in protein synthesis in neonatal and adult cardiac myocytes. Superresolution microscopy was used to examine the interaction of ribosomes and the SR in adult cardiac myocytes. ResultsQuantification of ER and SR-associated proteins of isolated ventricular cardiac myocytes showed that relative expression of ER/SR resident protein quality elements, as well as relative ribosome levels are decreased in adult cells, whereas SR-associated Ca2+ handling proteins increase. Immunocytoflourescence revealed that the membrane compartment that exists in early postpartum resembles mostly the ER and decreases in postnatal development. The SR is the main membrane network that exists in the adult cardiac myocytes, replacing the ER, in all but the perinuclear region. Immunocytoflourescence staining further indicated that both networks perform overlapping but distinct, specialized functions, such as localization of excitation-contraction coupling exclusively to the SR or initiation of secretion via the classical secretory pathway mainly from the ER. Ribosomes and mRNA were localized both in close proximity to the ER and the SR of adult ventricular cardiac myocytes. Superresolution microscopy confirmed that both the ER as well as the developed SR bind ribosomes and are direct sites of protein synthesis and protein homeostasis in adult cardiac myocytes. ConclusionOur findings suggest molecular differentiation and structural organization of the ER/SR in cardiac muscle development, resulting in the development of a protein synthesis network at the sarco/endoplasmic reticulum in adult cardiac myocytes.

cell biology↗