Biology subjects

Stein, W.

Publications and source records attributed to Stein, W..

11 recordsLinked to original sources

bioRxiv2026

Elevated precontractile calcium set point is sustained by limited PMCA1 export in dystrophic muscle

Duchenne muscular dystrophy is caused by loss of dystrophin and marked by elevated muscle calcium, usually attributed to contraction-induced membrane damage. Whether calcium rises before muscle can contract, and what sustains it, is unresolved. In Caenorhabditis elegans embryos and patient-derived human myoblasts, dystrophic muscle carries an elevated resting calcium set point before contraction is possible. Embryonic calcium is elevated 400 minutes after fertilization, before movement is detectable or the neuromuscular junction forms. Silencing the plasma membrane calcium pump gene mca-1 raises calcium in wild-type but not dystrophic animals, whereas silencing its paralogue raises it in both: the limitation is specific to that export route. Dystrophic human myoblasts have less pump; pump inhibition or silencing increases calcium preferentially in healthy cells, and raising pump expression lowers calcium most in dystrophic cells. Constrained calcium export is an early, continuing determinant of the dystrophic set point.

pathology↗

bioRxiv2025

Whole brain fluorescence imaging in Drosophila reveals spreading depression and its initiation, propagation, and resilience dynamics.

Spreading depression (SD) is a wave of neuronal hyperactivity followed by depolarization block that propagates through large brain regions and is associated with disorders such as migraine, stroke, and brain injury. The mechanisms that initiate SD and alter susceptibility to it remain incompletely understood. Here, we use whole-brain fluorescence imaging with genetically encoded pan-neuronal calcium and voltage sensors to observe SD in Drosophila melanogaster. We show that rapid cooling, a naturally occurring environmental condition, as well as elevated extracellular potassium reliably elicit SD in both adult and larval flies. SD was characterized by a rapid and large rise in intracellular calcium that was accompanied by neuronal depolarization and stark changes in the transperineuronal potential. In adults, SD occurred at 6.7 {+/-} 0.6{degrees}C (N=15) and in larvae at 6.0 {+/-} 0.3{degrees}C (N=30). SD initiation was not restricted to specific sites, but initiated at multiple, variable sites across and within individuals, with an average of 3.0 {+/-} 0.7 (N=8) initiation points per brain. In all cases, SD spread throughout large areas of the nervous system. In a high-throughput larval assays that allows the simultaneous monitoring of up to 16 animals with repeated cooling cycles, we demonstrate that single SD events are followed by a transient refractory period lasting up to 45 minutes, during which the threshold for subsequent SD was significantly elevated. This was the case in adult and larval brains of all developmental stages. The refractory effect was independent of neuronal depolarization, suggesting that homeostatic processes alter SD susceptibility following an initial SD event. Taken together, our findings demonstrate that SD initiation and propagation are not restricted to specific regions, neuronal populations, or developmental stages, and they reveal fundamental properties of adaptive changes to SD susceptibility in a genetically tractable model. Building upon the extensive genetic toolkit available in Drosophila, this work establishes the fly as a complementary model for understanding conserved cellular and circuit-level mechanisms of SD relevant to human neurological disorders.

neuroscience↗

bioRxiv2024

Effects of Martian magnetic and gravitational fields across multiple generations of the nematode C. elegans

AbstractsLife on Mars will require organisms to endure sustained exposure to reduced gravity and near-absent planetary magnetism, yet little is known about how these environmental factors may combine to potentially affect biology across generations. Here, we investigated the transgenerational effects of Martian gravity and magnetism on neuromuscular, sensory, and morphological function using Caenorhabditis elegans as a model. Animals were reared continuously under ground-based simulated Martian conditions across six generations and assessed using high-throughput behavioral and morphometric assays. Animals remained viable but acquired progressive transgenerational impairments across multiple functional domains. Swimming frequency showed immediate and severe deficits at all generations tested (Cohens d = 2.6-4.2), while chemotaxis deficits emerged more gradually, becoming significant by Generation 4. Morphological changes followed non-monotonic trajectories, with transient compensatory growth at Generation 4 followed by increased developmental variability at Generation 6. These differential patterns of impairment across neuromuscular, sensory, and developmental systems reveal domain-specific vulnerabilities to sustained Martian conditions. Critically, by Generation 6, Mars lineages exhibited a three-to-eight-fold increase in phenotypic variability, consistent with a loss of developmental canalization that could pose a more serious challenge for sustained colonization than simple mean fitness reductions.

physiology↗

bioRxiv2024

Muscular expression of pezo-1 differentially contributes to swimming and crawling production in the nematode C. elegans.

Mechanosensitive PIEZO ion channels are evolutionarily conserved proteins that are widely expressed in neuronal and muscular tissues. This study explores the role of the mechanoreceptor PEZO-1 in the body wall muscles of Caenorhabditis elegans, focusing on its influence on two locomotor behaviors, swimming and crawling. Using confocal imaging, we reveal that PEZO-1 localizes to the sarcolemma and plays a crucial role in modulating calcium dynamics that are important for muscle contraction. When we knocked down pezo-1 expression in striated muscles with RNA interference, calcium levels in head and tail muscles increased. While heightened, the overall trajectory of the calcium signal during the crawl cycle remained the same. While downregulation of pezo-1 led to an increase in crawling speed, it caused a reduction in swimming speed. Reduction in pezo-1 expression also resulted in the increased activation of the ventral tail muscles, and a disruption of dorsoventral movement asymmetry, a critical feature that enables propulsion in water. These alterations were correlated with impaired swimming posture and path curvature, suggesting that PEZO-1 has different functions during swimming and crawling.

biophysics↗

bioRxiv2023

The AMsh glia of C. elegans modulates the duration of touch-induced escape responses

Once considered mere structural support cells in the nervous system, glia have recently been demonstrated to play pivotal roles in sensorimotor processing and to directly respond to sensory stimuli. However, their response properties and contributions to sensory-induced behaviors remain little understood. In Caenorhabditis elegans, the amphid sheath glia (AMsh) directly respond to aversive odorants and mechanical stimuli, but their precise transduction machinery and their behavioral relevance remain unclear. We investigated the role of AMsh in mechanosensation and their impact on escape behaviors in C. elegans. We found that nose touch stimuli in immobilized animals induced a slow calcium wave in AMsh, which coincided with the termination of escape reversal behaviors. Genetic ablation of AMsh resulted in prolonged reversal durations in response to nose touch, but not to harsh anterior touch, highlighting the specificity of AMshs role in distinct escape behaviors. Mechanotransduction in AMsh requires the -tubulin MEC-12 and the ion channels ITR-1 and OSM-9, indicating a unique mechanosensory pathway that is distinct from the neighboring ASH neurons. We find that GABAergic signaling mediated by the GABA-A receptor orthologs LGC-37/8 and UNC-49 play a crucial role in modulating the duration of nose touch-induced reversals. We conclude that in addition to aversive odorant detection, AMsh mediate mechanosensation, play a pivotal role in terminating escape responses to nose touch, and provide a mechanism to maintain high sensitivity to polymodal sensory stimuli. SignificancePolymodal nociceptive sensory neurons have the challenge of multitasking across sensory modalities. They must respond to dangerous stimuli of one modality, but also adapt to repeated nonthreatening stimuli without compromising sensitivity to harmful stimuli from different modalities. Here we show that a pair of glia in the nematode C. elegans modulate the duration of nose-touch induced escape responses. We identify several molecules involved in the transduction of mechanical stimuli in these cells and show that they use the signaling molecule GABA to modulate neural function. We propose a mechanism through which these glia might function to maintain this polysensory neuron responsive to dangerous stimuli across different modalities.

neuroscience↗

bioRxiv2023

Neuromodulator-induced temperature robustness in a motor pattern: a comparative study between three decapod crustaceans

While temperature fluctuations pose significant challenges to the nervous system, many vital neuronal systems in poikilothermic animals function over a broad temperature range. Using the gastric mill pattern generator in the Jonah crab, we previously demonstrated that temperature-induced increases in leak conductance disrupt neuronal function and that neuropeptide modulation provides thermal protection. Here, we show that neuropeptide modulation also increases temperature robustness in Dungeness and Green crabs. Like in Jonah crabs, higher temperatures increased leak conductance in both species pattern-generating neuron LG and terminated rhythmic gastric mill activity. Likewise, increasing descending modulatory projection neuron activity or neuropeptide transmitter application rescued rhythms at elevated temperatures. However, decreasing input resistance using dynamic clamp only restored the rhythm in half of the experiments. Thus, neuropeptide modulation increased temperature robustness in both species, demonstrating that neuropeptide-mediated temperature compensation is not limited to one species, although the underlying cellular compensation mechanisms may be distinct. Summary statementThis study shows that the release of neuropeptide from modulatory projection neurons plays a crucial role in maintaining neuron and circuit function at elevated temperatures across crustacean species.

neuroscience↗

bioRxiv2021

The hyperpolarization-activated current shifts the dynamic range of a voltage-dependent electrical synapse

Like their chemical counterparts, electrical synapses show complex dynamics such as rectification and voltage dependence that interact with other electrical processes in neurons. The consequences arising from these interactions for the electrical behavior of the synapse, and the dynamics they create, remain largely unexplored. Using a voltage-dependent electrical synapse between a descending modulatory projection neuron (MCN1) and a motor neuron (LG) in the crustacean stomatogastric ganglion, we find that the influence of the hyperpolarization-activated inward current (Ih) is critical to the function of the electrical synapse. When we blocked Ih with CsCl, the apparent voltage dependence of the electrical synapse shifted by 18.7 mV to more hyperpolarized voltages, placing the dynamic range of the electrical synapse outside of the range of voltages used by the LG motor neuron (-60.2 mV - -44.9 mV). With dual electrode current- and voltage-clamp recordings, we demonstrate that this voltage shift is not due to a change in the properties of the gap junction itself, but is a result of a sustained effect of Ih on the presynaptic MCN1 axon terminal membrane potential. Ih-induced depolarization of the axon terminal membrane potential increased the electrical postsynaptic potentials and currents. With Ih present, the axon terminal resting membrane potential depolarized, shifting the dynamic range of the electrical synapse towards the functional range of the motor neuron. We thus demonstrate that the function of an electrical synapse is critically influenced by a voltage-dependent ionic current (Ih). New & NoteworthyElectrical synapses and voltage-gated ionic currents are often studied independently from one another, despite mounting evidence that their interactions can alter synaptic behavior. We show that the hyperpolarization-activated inward ionic current shifts the voltage dependence of an electrical synaptic transmission through its depolarizing effect on the membrane potential, enabling it to lie within the functional membrane potential range of a motor neuron. Thus, the electrical synapses function critically depends on the voltage-gated ionic current.

neuroscience↗

bioRxiv2021

Neuromodulation enables temperature robustness and coupling between fast and slow oscillator circuits in Cancer borealis

Acute temperature changes can disrupt neuronal activity and coordination with severe consequences for animal behavior and survival. Nonetheless, two rhythmic neuronal circuits in the crustacean stomatogastric ganglion (STG) and their coordination are maintained across a broad temperature range. However, it remains unclear how this temperature robustness is achieved. Here, we dissociate temperature effects on the rhythm generating circuits from those of upstream ganglia. We demonstrate that heat-activated factors extrinsic to the rhythm generators are essential to the slow gastric mill rhythms temperature robustness and contribute to the temperature response of the fast pyloric rhythm. The gastric mill rhythm crashed when only the STG circuits were heated. It could be restored when upstream ganglia were heated in addition, and the activity of the peptidergic modulatory projection neuron (MCN1) increased. Correspondingly, MCN1s neuropeptide transmitter stabilized the rhythm and maintained it over a broad temperature range. Extrinsic neuromodulation is thus essential for the oscillatory circuits in the STG and enables neural circuits to maintain function in temperature-compromised conditions. In contrast, integer coupling between pyloric and gastric mill rhythms was independent of whether extrinsic inputs and STG pattern generators were temperature-matched or not, demonstrating that the temperature robustness of the coupling is enabled by properties intrinsic to the rhythm generators. However, at near-crash temperature, integer coupling was maintained only in some animals but was absent in others. This was true despite regular rhythmic activity in all animals, supporting that degenerate circuit properties result in idiosyncratic responses to environmental challenges.

neuroscience↗

bioRxiv2021

Distinct mechanoreceptor pezo-1 isoforms modulate food intake in the nematode Caenorhabditis elegans

Two PIEZO mechanosensitive cation channels, PIEZO1 and PIEZO2, have been identified in mammals, where they are involved in numerous sensory processes. While structurally similar, PIEZO channels are expressed in distinct tissues and exhibit unique properties. How different PIEZOs transduce force, how their transduction mechanism varies, and how their unique properties match the functional needs of the tissues they are expressed in remain all-important unanswered questions. The nematode Caenorhabditis elegans has a single PIEZO ortholog (pezo-1) predicted to have twelve isoforms. These isoforms share many transmembrane domains but differ in those that distinguish PIEZO1 and PIEZO2 in mammals. We used transcriptional and translational reporters to show that putative promoter sequences immediately upstream of the start codon of long pezo-1 isoforms predominantly drive GFP expression in mesodermally derived tissues (such as muscle and glands). In contrast, sequences upstream of shorter pezo-1 isoforms resulted in GFP expression primarily in neurons. Putative promoters upstream of different isoforms drove GFP expression in different cells of the same organs of the digestive system. The observed unique pattern of complementary expression suggests that different isoforms could possess distinct functions within these organs. We used mutant analysis to show that pharyngeal muscles and glands require long pezo-1 isoforms to respond appropriately to the presence of food. The number of pezo-1 isoforms in C. elegans, their putative differential pattern of expression, and roles in experimentally tractable processes make this an attractive system to investigate the molecular basis for functional differences between members of the PIEZO family of mechanoreceptors.

genetics↗

bioRxiv2021

Interneuronal dynamics facilitate the initiation of cortical spreading depression

Cortical spreading depression (CSD) is thought to precede migraine attacks with aura and is characterized by a slowly traveling wave of inactivity through cortical pyramidal cells. During CSD, pyramidal cells experience hyperexcitation with rapidly increasing firing rates, major changes in electrochemistry, and ultimately spike block that propagates slowly across the cortex. While the identifying characteristic of CSD is the pyramidal cell hyperexcitation and subsequent spike block, it is currently unknown how the dynamics of the cortical microcircuits and inhibitory interneurons affect the initiation of CSD. We tested the contribution of cortical inhibitory interneurons to the initiation of spike block using a cortical microcircuit model that takes into account changes in ion concentrations that result from neuronal firing. Our results show that interneuronal inhibition provides a wider dynamic range to the circuit and generally improves stability against spike block. Despite these beneficial effects, strong interneuronal firing contributed to rapidly changing extracellular ion concentrations, which facilitated hyperexcitation and led to spike block first in the interneuron and then in the pyramidal cell. In all cases, a loss of interneuronal firing triggered pyramidal cell spike block. However, preventing interneuronal spike block was insufficient to rescue the pyramidal cell from spike block. Our data thus demonstrate that while the role of interneurons in cortical microcircuits is complex, they are critical to the initiation of pyramidal cell spike block and CSD. We discuss the implications that localized effects on cortical interneurons have beyond the isolated microcircuit.

neuroscience↗

bioRxiv2019

Unbiased analysis of magnetic orientation by C. elegans reveals the use of distinct turning strategies to align with magnetic fields and to adopt their preferred migratory direction.

To successfully navigate their surroundings, animals detect and orient to environmental stimuli possessing unique physical properties. Most animals can derive directional information from spatial or temporal changes in stimulus intensity (e.g. chemo- and thermo-taxis). However, some biologically relevant stimuli have constant intensity at most organismal scales. The gravitational and magnetic fields of the earth are examples of uniform stimuli that remain constant at most relevant scales. While devoid of information associated with intensity changes, the vectorial nature of these fields intrinsically encodes directional information. While much is known about behavioral strategies that exploit changes in stimulus intensity (gradients), less is understood about orientation to uniform stimuli. Nowhere is this truer than with magnetic orientation. While many organisms are known to orient to the magnetic field of the earth, how these animals extract information from the earths magnetic field remains unresolved.\n\nHere we use the nematode C. elegans to investigate behavioral strategies for orientation to magnetic fields, and compare our findings to the better characterized chemical and thermal orientation strategies. We used an unbiased cluster analysis to categorize, quantify, and compare behavioral components underlying different orientation strategies as a way to quantify and compare animal orientation to distinct stimuli. We find that in the presence of an earth-like magnetic field, worms perform acute angle turns (140-171{degrees}) that significantly improved their alignment with the direction of an imposed magnetic vector. In contrast, animals performed high amplitude turns (46-82{degrees}) that significantly increased alignment of their trajectory with the preferred migratory angle. We conclude that C. elegans orients to earth-strength magnetic fields using two independent behavioral strategies, in contrast to orientation strategies to graded stimuli. Understanding how C. elegans detects and orients to magnetic fields will provide useful insight into how many species across taxa accomplish this fascinating sensory feat.

animal behavior and cognition↗