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Biology subjects

Schulze, H.

Publications and source records attributed to Schulze, H..

17 recordsLinked to original sources

Chronic stress does not induce behavioural signs of tinnitus or cochlear synaptopathy in Mongolian gerbils

Tinnitus is commonly associated with psychosocial stress. However, it is unclear whether stress alone is sufficient to induce neural alterations resulting in tinnitus. In this study, we investigated the causal role of stress in tinnitus generation by experimentally isolating stress exposure as the sole independent variable in a well-established Mongolian gerbil model. Animals were subjected to repeated, inescapable electric foot shocks over three weeks to create a chronic, repeated stress paradigm. Behavioural, endocrine and histological measures were combined to assess tinnitus perception, stress system activation and cochlear synaptopathy. Repeated stress exposure reliably activated the endocrine stress response, as indicated by transient increases in serum cortisol following specific stress sessions. Basal cortisol levels returned to baseline values after the stress period. Behavioural assessment using gap-prepulse inhibition of the acoustic startle reflex revealed only isolated cases of behavioural signs of tinnitus, which occurred at a frequency consistent with false-positive detection and were evenly distributed between stress-exposed and control animals. Histological analyses revealed that ribbon synapse counts were preserved across the cochlea, with no evidence of stress-induced synaptopathy. No systematic relationships were observed between endocrine activation, behavioural outcomes and the number of inner hair cell ribbon synapses. Taken together, these findings suggest that, in the absence of acoustic trauma, repeated chronic stress is not sufficient to induce tinnitus or inner hair cell synaptopathy. These results argue against a primary causal role of stress alone in tinnitus generation and instead support models in which stress modulates symptom expression or perceptual salience in the context of pre-existing auditory dysfunction.

neuroscience↗

mCytoCounter, a deployable somatic cell counting device for the assessment of mastitis in dairy farms

Bovine mastitis is a recurring disease among herds that is often caused by unhygienic farm conditions, resulting in bacterial invasion of the udder. It is a leading cause of financial losses for dairy farmers, largely due to the widespread use of antimicrobials for treatment, which in turn fuels the development of drug-resistant bacteria. Somatic cell count (SCC) is a widely used quantitative method for diagnosing sub-clinical and clinical mastitis. We have developed a rapid and quantitative microfluidic device that enables the quantification of somatic cells in milk, thereby aiding in the diagnosis of bovine mastitis. Our platform features a microfluidic cartridge for capturing somatic cells, which is inserted into an in-house device called the mCytoCounter. The device reads the fluorescence emission signals from the somatic cells captured on the membrane in 2 minutes, with a total turnaround time of 15 minutes. The mCytoCounter is an automated and deployable system that can be used in dairy farms with minimal training. We analysed the performance of mCytoCounter with milk samples and benchmarked it against the SCC using a fluorescence microscope with a correlation coefficient of 0.97. The mCytoCounter has a detection range of 103 to 106 SCC/mL, which covers the required cell number range to discriminate between healthy, subclinical, and clinical mastitis.

immunology↗

Thrombo-inflammation analyzed in a validated seven-layer platelet decision model: cellular decisions are tough problems fast and heuristically solved

Decisions in biology happen fast and are driven by evolution to optimize survival chances. In platelets, this is achieved by organizing signaling cascades into rapid decision-funnels with modulatory crosstalk. We show that network decision processes underlying cellular decisions are tough to solve (equivalent to classical satisfiability problems, SAT). Hence, heuristics, modular decision-making, and decision funnels are required for efficient decisions. We establish this using a seven-layer platelet decision network that agrees well with all available genetic and functional experimental data. Platelet decision cascades are robust to perturbations: For example, receptors such as TRPM7 modulate platelet activity. However, knockouts of the receptors still leave platelets reactive overall. Dynamic control resolves relaying functions from kinases to cytoskeleton alterations. This allows fast execution of platelet shape change or aggregation. Stress conditions can shift platelet decision funnels towards constant activation of aggregation or immune signaling, causing thrombosis or thrombo-inflammation. Based on the network dynamics, we conclude that platelets pragmatically resolve the complex (non-polynomial (NP)) cellular decision problems by using a similar relaxation to those proposed in mathematics - many different configurations end up in similar states. Metamathematical considerations (no mathematical proof) suggest that NP problems are more complex then P problems. One sentence abstractWe show that cellular decision problems like the platelet signaling cascade may need unexpectedly long to solve but in general, they are efficiently solved using heuristics ("decision funnels"), implying fast decisions but the risk of chronic stress and inflammation.

bioinformatics↗

Behavioral assessment of Zwicker tone percepts in rodents

The Zwicker tone illusion can serve as an interesting model for acute tinnitus, an auditory phantom percept still not fully understood. Recent mechanistic models suggest that the underlying neural mechanisms of both percepts are similar. However, to date it is not clear if animals do perceive the Zwicker tone at all, as up to now no behavioral paradigms are available to objectively assess the presence of this phantom percept. Here we introduce, for the first time, a modified version of the gap pre-pulse inhibition of the acoustic startle reflex (GPIAS) paradigm - usually used to assess the presence of a tinnitus percept in animals - to test if it is possible to induce a Zwicker tone percept in our rodent model, the Mongolian gerbil. Furthermore, we developed a new aversive conditioning shuttle box learning paradigm and compare the two approaches. We found a significant increase in the GPIAS effect when presenting a notched noise compared to flat white noise gap pre-pulse inhibition, indicating that the animals actually perceived a Zwicker tone. However, in the aversive conditioning learning paradigm, no clear effect could be observed in the discrimination performance of the tested animals. When investigating the CR+ responses, an effect of a possible Zwicker tone percept can be seen, i.e. animals show identical behavior as if a pure tone was presented, but the paradigm needs to be further improved.

neuroscience↗

Fusing spheroids to aligned μ-tissues in a Heart-on-Chip featuring oxygen sensing and electrical pacing capabilities

Over the last decade Organ-on-Chip (OOC) emerged as a promising technology for advanced in vitro models, recapitulating key physiological cues. OOC approaches tailored for cardiac tissue engineering resulted in a variety of platforms, some of which integrate stimulation or probing capabilities. Due to manual handling processes, however, a large-scale standardized and robust tissue generation, applicable in an industrial setting, is still out of reach. Here, we present a novel cell injection and tissue generation concept relying on spheroids, which can be produced in large quantities and uniform size from induced pluripotent stem cell-derived human cardiomyocytes. Hydrostatic flow transports and accumulates spheroids in dogbone-shaped cultivation chambers, which subsequently fuse and form aligned, contracting cardiac muscle fibers. Furthermore, we demonstrate electrical stimulation capabilities by utilizing fluidic media connectors as electrodes and provide the blueprint of a low-cost, open-source, scriptable pulse generator. We report on a novel integration strategy of optical O2 sensor spots into resin-based microfluidic systems, enabling in situ determination of O2 partial pressures. Finally, proof-of-concept demonstrating electrical stimulation combined with in situ monitoring of metabolic activity in cardiac tissues is provided. The developed system thus opens the door for advanced OOCs integrating biophysical stimulation as well as probing capabilities and serves as blueprint for the facile and robust generation of high density microtissues in microfluidic modules amenable for scale-up and automation.

bioengineering↗

G6b-B regulates an essential step in megakaryocyte maturation

G6b-B is a megakaryocyte lineage-specific immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor, essential for platelet homeostasis. Mice with a genomic deletion of the entire Mpig6b locus develop severe macrothrombocytopenia and myelofibrosis, which is reflected in humans with null-mutations in MPIG6B. The current model proposes that megakaryocytes lacking G6b-B develop normally, while proplatelet release is hampered, but the underlying molecular mechanism remains unclear. Here, we report on a spontaneous recessive single nucleotide mutation in C57BL/6 mice, localized within the intronic region of the Mpig6b locus that abolishes G6b-B expression and reproduces macrothrombocytopenia, myelofibrosis and osteosclerosis. As the mutation is based on a single nucleotide exchange, Mpig6bmut mice represent an ideal model to study the role of G6b-B. Megakaryocytes from these mice were smaller in size, displayed a less developed demarcation membrane system and reduced expression of receptors. RNA sequencing revealed a striking global reduction in the level of megakaryocyte-specific transcripts, in conjunction with decreased protein levels of the transcription factor GATA-1, and impaired thrombopoietin signaling. The reduced number of mature MKs in the bone marrow was corroborated on a newly developed Mpig6b null mouse strain. Our findings highlight an unexpected essential role of G6b-B in the early differentiation within the megakaryocytic lineage.

cell biology↗

How biological cells including platelets and megakaryocytes decide complex problems fast but risky

Short AbstractMathematical decision processes are accurate but sometimes take very long time or simply do not happen. Decisions in biology happen fast and driven by evolution, optimizing survival chances. This results in stochastic decisions with on average good adaptation to the environment but an inherent risk of individual errors e.g. developing cancer during cell regeneration. We calculate and show in platelets and megakaryocytes how cellular decision processes increases risk for errors and inflammation. Short cut solutions adapted from nature improve computer strategies for protein folding and network decision processes. Complex problems are not always solved in foreseeable time, instead the fast solutions in biology speed up errors everywhere including biochemical aging of blood vessels, misfolded proteins, mis-programmed cells, cancer and heart failure. One sentence abstractWe investigate in biological networks how complex decision problems are mastered not by an accurate but unforeseeable long mathematical search but rather pragmatic and fast, with an inherent risk of error, a basis for inflammation and cancer.

bioinformatics↗

Sleep as a random walk - A superstatistical analysis of EEG data across sleep stages

In clinical practice, human sleep is classified into stages, each associated with different levels of muscular activity and marked by characteristic patterns in the EEG signals. It is however unclear whether this subdivision into discrete stages with sharply defined boundaries is truly reflecting the dynamics of human sleep. To address this question, we consider one-channel EEG signals as heterogeneous random walks: stochastic processes controlled by hyper-parameters that are themselves time-dependent. We first demonstrate the heterogeneity of the random process by showing that each sleep stage has a characteristic distribution and temporal correlation function of the raw EEG signals. Next, we perform a superstatistical analysis by computing hyper-parameters, such as the standard deviation, kurtosis and skewness of the raw signal distributions, within subsequent 30-second epochs. It turns out that also the hyper-parameters have characteristic, sleep-stage-dependent distributions, which can be exploited for a simple Bayesian sleep stage detection. Moreover, we find that the hyper-parameters are not piece-wise constant, as the traditional hypnograms would suggest, but show rising or falling trends within and across sleep stages, pointing to an underlying continuous rather than subdivided process that controls human sleep.

neuroscience↗

Analysis and Visualization of Sleep Stages based on Deep Neural Networks

Automatic sleep stage scoring based on deep neural networks has come into focus of sleep researchers and physicians, as a reliable method able to objectively classify sleep stages, would save human resources and thus would simplify clinical routines. Due to novel open-source software libraries for Machine Learning in combination with enormous progress in hardware development in recent years a paradigm shift in the field of sleep research towards automatic diagnostics could be observed. We argue that modern Machine Learning techniques are not just a tool to perform automatic sleep stage classification but are also a creative approach to find hidden properties of sleep physiology. We have already developed and established algorithms to visualize and cluster EEG data, in a way so that we can already make first assessments on sleep health in terms of sleep-apnea and consequently daytime vigilance. In the following study, we further developed our method by the innovative approach to analyze cortical activity during sleep by computing vectorial cross-correlations of different EEG channels represented by hypnodensity graphs. We can show that this measure serves to estimate the period length of sleep cycles and thus can help to find disturbances due to pathological conditions.

neuroscience↗

The Stochastic Resonance model of auditory perception: A unified explanation of tinnitus development, Zwicker tone illusion, and residual inhibition.

Stochastic Resonance (SR) has been proposed to play a major role in auditory perception, and to maintain optimal information transmission from the cochlea to the auditory system. By this, the auditory system could adapt to changes of the auditory input at second or even sub-second timescales. In case of reduced auditory input, somatosensory projections to the dorsal cochlear nucleus would be disinhibited in order to improve hearing thresholds by means of SR. As a side effect, the increased somatosensory input corresponding to the observed tinnitus-associated neuronal hyperactivity is then perceived as tinnitus. In addition, the model can also explain transient phantom tone perceptions occurring after ear plugging, or the Zwicker tone illusion. Vice versa, the model predicts that via stimulation with acoustic noise, SR would not be needed to optimize information transmission, and hence somatosensory noise would be tuned down, resulting in a transient vanishing of tinnitus, an effect referred to as residual inhibition.

neuroscience↗

Intrinsic noise improves speech recognition in a computational model of the auditory pathway

Noise is generally considered to harm information processing performance. However, in the context of stochastic resonance, noise has been shown to improve signal detection of weak subthreshold signals, and it has been proposed that the brain might actively exploit this phenomenon. Especially within the auditory system, recent studies suggest that intrinsic noise plays a key role in signal processing and might even correspond to increased spontaneous neuronal firing rates observed in early processing stages of the auditory brain stem and cortex after hearing loss. Here we present a computational model of the auditory pathway based on a deep neural network, trained on speech recognition. We simulate different levels of hearing loss and investigate the effect of intrinsic noise. Remarkably, speech recognition after hearing loss actually improves with additional intrinsic noise. This surprising result indicates that intrinsic noise might not only play a crucial role in human auditory processing, but might even be beneficial for contemporary machine learning approaches.

neuroscience↗

Acute and long-term circuit-level effects in the auditory cortex after sound trauma

Harmful environmental sounds are a prevailing source for chronic hearing impairments, including noise induced hearing loss, hyperacusis, or tinnitus. How these symptoms are related to pathophysiological damage to the sensory receptor epithelia and its effects along the auditory pathway, such as functional reorganizations in the auditory cortex (ACx), have been documented in numerous studies. An open question concerns the temporal evolution of maladaptive changes after damage and their manifestation in the balance between afferent thalamocortical input and corticocortical input to the ACx. To address this, we investigated the loci of plastic reorganizations across the tonotopic axis of the auditory cortex of male Mongolian gerbils (Meriones unguiculatus) acutely after a sound trauma and after several weeks. We used a laminar residual current-source density analysis to dissociate adaptations of intracolumnar input and horizontally relayed corticocortical input to synaptic populations across cortical layers in ACx. A pure tone-based sound trauma caused acute changes of subcortical inputs and corticocortical inputs at all tonotopic regions, particularly showing a broad elimination of tone-evoked inputs at tonotopic regions with a pre-trauma best frequency between 2-8 kHz. At other cortical sites, the overall columnar activity acutely decreased, while relative contributions of lateral corticocortical inputs increased. After 4-6 weeks, cortical activity to the altered sensory inputs showed a general increase of local thalamocortical input reaching levels higher than before the trauma. Hence, our results suggest a detailed mechanism for overcompensation of altered frequency input in the auditory cortex that relies on a changing balancing of thalamocortical and intracortical input and is confined to the spectral neighborhood of the trauma frequency. Significance statementHarmful noise exposure is a major anthropogenic cause of hearing disorders and is becoming an ever-increasing burden for human health and society. Damage to the sensory epithelia elicited by harmful sounds can subsequently lead to chronic hearing loss, hyperacusis or tinnitus. We still lack an understanding of the pathophysiological plastic processes and their evolution, particularly at the circuit level of the auditory cortex (ACx), which is fundamentally involved in auditory perception. We demonstrate that plastic changes in ACx after noise induced hearing loss (NIHL) occur over several weeks, and that changes in intracortical functional connectivity compensate the acute effects in the deafferentiated subcortical inputs. Such long-term changes may underlie the temporal evolution of hearing impairments or phantom sounds after NIHL.

neuroscience↗

Impact of Itga2-Gp6-double collagen receptor deficient mice for bone marrow megakaryocytes and platelets

The two main collagen receptors on platelets, GPVI and integrin 2{beta}1, play an important role for the recognition of exposed collagen at sites of vessel injury, which leads to platelet activation and subsequently stable thrombus formation. Both receptors are already expressed on megakaryocytes, the platelet forming cells within the bone marrow. Megakaryocytes are in permanent contact with collagen filaments in the marrow cavity and at the basal lamina of sinusoids without obvious preactivation. The role of both collagen receptors for megakaryocyte maturation and thrombopoiesis is still poorly understood. To investigate the function of both collagen receptors, we generated mice that are double deficient for Gp6 and Itga2. Flow cytometric analyses revealed that the deficiency of both receptors had no impact on platelet number and the expected lack in GPVI responsiveness. Integrin activation and degranulation ability was comparable to wildtype mice. By immunofluorescence microscopy, we could demonstrate that double-deficient megakaryocytes were overall normally distributed within the bone marrow. We found megakaryocyte count and size to be normal, the localization within the bone marrow, the degree of maturation, as well as their association to sinusoids were also unaltered. However, the contact of megakaryocytes to collagen type I filaments was decreased at sinusoids compared to wildtype mice, while the interaction to type IV collagen was unaffected. Our results imply that GPVI and 2{beta}1 have no influence on the localization of megakaryocytes within the bone marrow, their association to the sinusoids or their maturation. The decreased contact of megakaryocytes to collagen type I might at least partially explain the unaltered platelet phenotype in these mice, since proplatelet formation is mediated by these receptors and their interaction to collagen. It is rather likely that other compensatory signaling pathways and receptors play a role that needs to be elucidated.

cell biology↗

Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B

The immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor G6b-B is critical for platelet production and activation, loss of which results in severe macrothrombocytopenia and aberrant platelet function in mice and humans. Using a combination of immunohistochemistry, affinity chromatography and proteomics, we identified the extracellular matrix heparan sulfate (HS) proteoglycan perlecan as a G6b-B binding partner. Subsequent in vitro biochemical studies and a cell-based genetic screen demonstrated that the interaction is specifically mediated by the HS chains of perlecan. Biophysical analysis revealed that heparin forms a high-affinity complex with G6b-B and mediates dimerization. Using platelets from humans and genetically-modified mice, we demonstrate that binding of G6b-B to HS and multivalent heparin inhibits platelet and megakaryocyte function by inducing downstream signaling via the tyrosine phosphatases Shp1 and Shp2. Our findings provide novel insights into how G6b-B is regulated and contribute to our understanding of the interaction of megakaryocytes and platelets with glycans.

biochemistry↗

Exercise restores chloride homeostasis and decreases spasticity through the BDNF-KCC2 pathway after chronic SCI

Activity-based therapies are routinely integrated in rehabilitation programs to induce repetitive activation of the neuromuscular system and facilitate functional recovery after spinal cord injury (SCI). Among the beneficial effects of physical therapy is a reduction of hyperreflexia and spasticity in SCI individuals, but the precise mechanism by which exercise regulates spinal networks and facilitate recovery remains elusive. Spasticity is a debilitating condition that affects ~ 75% of the SCI population and interferes with residual motor function. Current pharmacological treatments not only have serious side effects but also actively depress spinal excitability and interferes with motor recovery. Understanding how activity-based therapies contribute to decrease spasticity will help identify critical pharmacological targets and optimize rehabilitation programs.\n\nKCC2, a neuron-specific Cl- extruder, is critical to the maintenance of [Cl-]i and its downregulation after SCI leads to a shift in chloride homeostasis that contributes to develop spasticity. We have shown in earlier studies that not only exercise promotes reflex modulation but also restores KCC2 expression in motoneurons. KCC2 is dynamically modulated by several signaling pathways, the most prevalent being BDNF-TrkB. Interestingly, activity-dependent processes triggered by exercise include an increase in the expression of BDNF in the lumbar spinal cord. However, whether the increase in KCC2 contributes to functional recovery and rely on BDNF activity have not been established. Our objective was to determine 1) whether the activity-dependent upregulation of KCC2 contributes to decrease spasticity after SCI; 2) if BDNF regulates KCC2 expression in an activity-dependent manner.\n\nUsing a model of complete SCI, we investigated this possible causal effect by intrathecally delivering VU0240551, a specific KCC2 blocker, or TrkB-IgG, a BDNF scavenger. Drugs were specifically delivered during the daily rehabilitation sessions to transiently prevent KCC2/BDNF activity. We provide evidence that the beneficial effect of exercise on functional recovery relies on a BDNF-dependent increase in KCC2 expression on motoneurons and the restoration of endogenous inhibition to a mature state. We identify, for the first time, that the increase in KCC2 activity with activity-based therapies functionally contributes to H-reflex recovery and critically depends on BDNF activity. This provides a new perspective on our understanding of how exercise impact hyperreflexia by identifying the biological basis of recovery of function. Acting directly on chloride homeostasis through BDNF to restore endogenous inhibition rather than actively depress excitability can diminish the reduction in motor output associated with the current pharmacological management of SCI and improve the outcome of rehabilitation programs.

neuroscience↗

Weight statistics controls dynamics in recurrent neural networks

Recurrent neural networks are complex non-linear systems, capable of ongoing activity in the absence of driving inputs. The dynamical properties of these systems, in particular their long-time attractor states, are determined on the microscopic level by the connection strengths wij between the individual neurons. However, little is known to which extent network dynamics is tunable on a more coarse-grained level by the statistical features of the weight matrix. In this work, we investigate the dynamical impact of three statistical parameters: density (the fraction of non-zero connections), balance (the ratio of excitatory to inhibitory connections), and symmetry (the fraction of neuron pairs with wij = wji). By computing a phase diagram of network dynamics, we find that balance is the essential control parameter: Its gradual increase from negative to positive values drives the system from oscillatory behavior into a chaotic regime, and eventually into stationary fix points. Only directly at the border of the chaotic regime do the neural networks display rich but regular dynamics, thus enabling actual information processing. These results suggest that the brain, too, is fine-tuned to the edge of chaos by assuring a proper balance between excitatory and inhibitory neural connections.\n\nAuthor summaryComputations in the brain need to be both reproducible and sensitive to changing input from the environment. It has been shown that recurrent neural networks can meet these simultaneous requirements only in a particular dynamical regime, called the edge of chaos in non-linear systems theory. Here, we demonstrate that recurrent neural networks can be easily tuned to this critical regime of optimal information processing by assuring a proper ratio of excitatory and inhibitory connections between the neurons. This result is in line with several micro-anatomical studies of the cortex, which frequently confirm that the excitatory-inhibitory balance is strictly conserved in the cortex. Furthermore, it turns out that neural dynamics is largely independent from the total density of connections, a feature that explains how the brain remains functional during periods of growth or decay. Finally, we find that the existence of too many symmetric connections is detrimental for the above mentioned critical dynamical regime, but maybe in turn useful for pattern completion tasks.

neuroscience↗

Tinnitus development is associated with synaptopathy of inner hair cells in Mongolian gerbils

Human hearing loss (HL) is often accompanied by comorbidities like tinnitus which is affecting up to 15% of the adult population. Rodent animal studies could show that tinnitus may not only be a result of apparent HL due to cochlear hair cell damage but can also be a consequence of synaptopathy at the inner hair cells (IHC) already induced by moderate sound traumata. Here we investigate synaptopathy previously shown in mice in our animal model, the Mongolian gerbil, and relate it to behavioral signs of tinnitus. Tinnitus was induced by a mild monaural acoustic trauma leading to monaural noise induced HL in the animals, quantified by auditory brainstem response (ABR) audiometry. Behavioral signs of tinnitus percepts were detected by measurement of prepulse inhibition of the acoustic startle response in a gap-noise paradigm. 14 days after trauma, the cochleae of both ears were isolated and IHC synapses were counted within several spectral regions of the cochlea. Behavioral signs of tinnitus were only found in animals with IHC synaptopathy, independent of type of HL. On the other hand, animals with apparent HL but without behavioral signs of tinnitus showed a reduction in amplitudes of ABR waves I&II but no significant changes in the number of synapses at the IHC. We conclude - in line with the literature - that HL is caused by damage to the IHC or by other reasons but that the development of tinnitus, at least in our animal model, is closely linked to synaptopathy at the IHC.

neuroscience↗