Myelin plasticity in the ventral tegmental area is required for opioid reward.

All drugs of abuse induce long-lasting changes in synaptic transmission and neural circuit function that underlie substance-use disorders1,2. Another recently appreciated mechanism of neural circuit plasticity is mediated through activity-regulated changes in myelin that can tune circuit function and influence cognitive behaviour3-7. Here we explore the role of myelin plasticity in dopaminergic circuitry and reward learning. We demonstrate that dopaminergic neuronal activity-regulated myelin plasticity is a key modulator of dopaminergic circuit function and opioid reward. Oligodendroglial lineage cells respond to dopaminergic neuronal activity evoked by optogenetic stimulation of dopaminergic neurons, optogenetic inhibition of GABAergic neurons, or administration of morphine. These oligodendroglial changes are evident selectively within the ventral tegmental area but not along the axonal projections in the medial forebrain bundle nor within the target nucleus accumbens. Genetic blockade of oligodendrogenesis dampens dopamine release dynamics in nucleus accumbens and impairs behavioural conditioning to morphine. Taken together, these findings underscore a critical role for oligodendrogenesis in reward learning and identify dopaminergic neuronal activity-regulated myelin plasticity as an important circuit modification that is required for opioid reward.

Controlling heme redox properties in peptide amphiphile fibers with sequence and heme loading ratio.

Controlling the reduction midpoint potential of heme B is a key factor in many bioelectrochemical reactions, including long-range electron transport. Currently, there are a number of globular model protein systems to study this biophysical parameter; however, there are none for large polymeric protein model systems (e.g., the OmcS protein from G. sulfurreducens). Peptide amphiphiles, short peptides with a lipid tail that polymerize into fibrous structures, fill this gap. Here, we show a peptide amphiphile model system where one can tune the electrochemical potential of heme B by changing the loading ratio and peptide sequence. Changing the loading ratio resulted in the most significant increase, with values as high as -22 mV down to -224 mV. Circular dichroism spectra of certain sequences show Cotton effects at lower loading ratios that disappear as more heme B is added, indicating an ordered environment that becomes disrupted if heme B is overpacked. These findings can contribute to the design of functional self-assembling biomaterials.

A pathogenic DYT-THAP1 dystonia mutation causes hypomyelination and loss of YY1 binding.

Dystonia is a disabling disease that manifests as prolonged involuntary twisting movements. DYT-THAP1 is an inherited form of isolated dystonia caused by mutations in THAP1 encoding the transcription factor THAP1. The phe81leu (F81L) missense mutation is representative of a category of poorly understood mutations that do not occur on residues critical for DNA binding. Here, we demonstrate that the F81L mutation (THAP1F81L) impairs THAP1 transcriptional activity and disrupts CNS myelination. Strikingly, THAP1F81L exhibits normal DNA binding but causes a significantly reduced DNA binding of YY1, its transcriptional partner that also has an established role in oligodendrocyte lineage progression. Our results suggest a model of molecular pathogenesis whereby THAP1F81L normally binds DNA but is unable to efficiently organize an active transcription complex.

Genome-Informed Trophic Classification and Functional Characterization of Virulence Proteins from the Maize Tar Spot Pathogen Phyllachora maydis.

Phyllachora maydis is an ascomycete foliar fungal pathogen and the causal agent of tar spot in maize. Although P. maydis is considered an economically important foliar pathogen of maize, our general knowledge of the trophic lifestyle and functional role of effector proteins from this fungal pathogen remains limited. Here, we utilized a genome-informed approach to predict the trophic lifestyle of P. maydis and functionally characterized a subset of candidate effectors from this fungal pathogen. Leveraging the most recent P. maydis genome annotation and the CATAStrophy pipeline, we show that this fungal pathogen encodes a predicted carbohydrate-active enzymes (CAZymes) repertoire consistent with that of biotrophs. To investigate fungal pathogenicity, we selected 18 candidate effector proteins that were previously shown to be expressed during primary disease development. We assessed whether these putative effectors share predicted structural similarity with other characterized fungal effectors and determined whether any suppress plant immune responses. Using AlphaFold2 and Foldseek, we showed that one candidate effector, PM02_g1115, adopts a predicted protein structure similar to that of an effector from Verticillium dahlia. Furthermore, transient expression of candidate effector-fluorescent protein fusions in Nicotiana benthamiana revealed two putative effectors, PM02_g378 and PM02_g2610, accumulated predominantly in the cytosol, and three candidate effectors, PM02_g1115, PM02_g7882, and PM02_g8240, consistently attenuated chitin-mediated reactive oxygen species production. Collectively, the results presented herein provide insights into the predicted trophic lifestyle and putative functions of effectors from P. maydis and will likely stimulate continued research to elucidate the molecular mechanisms used by P. maydis to induce tar spot.

THAP1 modulates oligodendrocyte maturation by regulating ECM degradation in lysosomes.

Mechanisms controlling myelination during central nervous system (CNS) maturation play a pivotal role in the development and refinement of CNS circuits. The transcription factor THAP1 is essential for timing the inception of myelination during CNS maturation through a cell-autonomous role in the oligodendrocyte lineage. Here, we demonstrate that THAP1 modulates the extracellular matrix (ECM) composition by regulating glycosaminoglycan (GAG) catabolism within oligodendrocyte progenitor cells (OPCs). Thap1-/- OPCs accumulate and secrete excess GAGs, inhibiting their maturation through an autoinhibitory mechanism. THAP1 controls GAG metabolism by binding to and regulating the GusB gene encoding ß-glucuronidase, a GAG-catabolic lysosomal enzyme. Applying GAG-degrading enzymes or overexpressing ß-glucuronidase rescues Thap1-/- OL maturation deficits in vitro and in vivo. Our studies establish lysosomal GAG catabolism within OPCs as a critical mechanism regulating oligodendrocyte development.

DLK signaling in axotomized neurons triggers complement activation and loss of upstream synapses.

Axotomized spinal motoneurons (MNs) lose presynaptic inputs following peripheral nerve injury; however, the cellular mechanisms that lead to this form of synapse loss are currently unknown. Here, we delineate a critical role for neuronal kinase dual leucine zipper kinase (DLK)/MAP3K12, which becomes activated in axotomized neurons. Studies with conditional knockout mice indicate that DLK signaling activation in injured MNs triggers the induction of phagocytic microglia and synapse loss. Aspects of the DLK-regulated response include expression of C1q first from the axotomized MN and then later in surrounding microglia, which subsequently phagocytose presynaptic components of upstream synapses. Pharmacological ablation of microglia inhibits the loss of cholinergic C boutons from axotomized MNs. Together, the observations implicate a neuronal mechanism, governed by the DLK, in the induction of inflammation and the removal of synapses.

Cholinergic Neuronal Activity Promotes Diffuse Midline Glioma Growth through Muscarinic Signaling.

Neuronal activity promotes the proliferation of healthy oligodendrocyte precursor cells (OPC) and their malignant counterparts, gliomas. Many gliomas arise from and closely resemble oligodendroglial lineage precursors, including diffuse midline glioma (DMG), a cancer affecting midline structures such as the thalamus, brainstem and spinal cord. In DMG, glutamatergic and GABAergic neuronal activity promotes progression through both paracrine signaling and through bona-fide neuron-to-glioma synapses. However, the putative roles of other neuronal subpopulations - especially neuromodulatory neurons located in the brainstem that project to long-range target sites in midline anatomical locations where DMGs arise - remain largely unexplored. Here, we demonstrate that the activity of cholinergic midbrain neurons modulates both healthy OPC and malignant DMG proliferation in a circuit-specific manner at sites of long-range cholinergic projections. Optogenetic stimulation of the cholinergic pedunculopontine nucleus (PPN) promotes glioma growth in pons, while stimulation of the laterodorsal tegmentum nucleus (LDT) facilitates proliferation in thalamus, consistent with the predominant projection patterns of each cholinergic midbrain nucleus. Reciprocal signaling was evident, as increased activity of cholinergic neurons in the PPN and LDT was observed in pontine DMG-bearing mice. In co-culture, hiPSC-derived cholinergic neurons form neuron-to-glioma networks with DMG cells and robustly promote proliferation. Single-cell RNA sequencing analyses revealed prominent expression of the muscarinic receptor genes CHRM1 and CHRM3 in primary patient DMG samples, particularly enriched in the OPC-like tumor subpopulation. Acetylcholine, the neurotransmitter cholinergic neurons release, exerts a direct effect on DMG tumor cells, promoting increased proliferation and invasion through muscarinic receptors. Pharmacological blockade of M1 and M3 acetylcholine receptors abolished the activity-regulated increase in DMG proliferation in cholinergic neuron-glioma co-culture and in vivo. Taken together, these findings demonstrate that midbrain cholinergic neuron long-range projections to midline structures promote activity-dependent DMG growth through M1 and M3 cholinergic receptors, mirroring a parallel proliferative effect on healthy OPCs.

A Neuro-Integrative Assessment of Perceptual-Motor Performance and Wellness in ROTC Cadets.

Resting heart rate variability (HRV) may be a useful index of both brain-based executive function and general health. Our purpose in this study was to quantify relationships among HRV, perceptual-motor performance metrics, and wellness survey responses. A cohort of 32 male Reserve Officer Training Corp (ROTC) cadets completed a dual-task upper extremity reaction time (UERT) test, two tests of whole-body reactive agility, and a 10-item wellness survey that produced a 0-100 Overall Wellness Index (OWI). We averaged participants' resting HRV measurements twice per week over 10 weeks to derive an intra-individual grand mean (HRV-IIGM) and over a series of days we calculated an intra-individual coefficient of variation (HRV-IICV). We used median values for the two HRV metrics (HRV-IIGM and HRV-IICV) to separate the cadets into equal-sized high and low HRV groups to form the dependent variable for logistic regression analyses. We found a significant inverse relationship between HRV-IIGM and HRV-IICV (r = -0.723, p < .001). Differences in UERT in the left versus right visual hemifields (L-R Diff) and OWI scores were strongly related to both HRV-IIGM ≤ 4.49 and HRV-IICV ≥ 6.95%. Logistic regression models that included L-R Diff and OWI showed 71% classification accuracy for HRV-IIGM (Model χ2 [2] = 12.47, p = .002, Nagelkerke R2 = 0.430) and 81% classification accuracy for HRV-IICV (Model χ2 [2] = 14.88, p = .001, Nagelkerke R2 = 0.496). These findings suggest that resting HRV, perceptual-motor efficiency, and overall wellness are highly interrelated, supporting a multi-factor biopsychosocial assessment to guide the design and implementation of interventions to maximize operational effectiveness for ROTC cadets and other military personnel.

Breaking away: violation of distortion emission phase-frequency invariance at low frequencies.

The phase versus frequency function of the distortion product otoacoustic emission (DPOAE) at 2f(1) - f(2) is approximately invariant at frequencies above 1.5 kHz in human subjects when recorded with a constant f(2)/f(1). However, a secular break from this invariance has been observed at lower frequencies where the phase-gradient becomes markedly steeper. Apical DPOAEs, such as 2f(1) - f(2), are known to contain contributions from multiple sources. This experiment asked whether the phase behavior of the ear canal DPOAE at low frequencies is driven by the phase of the component from the distortion product (DP) region at 2f(1) - f(2), which exhibits rapid phase accumulation. Placing a suppressor tone close in the frequency to 2f(1) - f(2) reduced the contribution of this component to the ear canal DPOAE in normal-hearing adult human ears. When the contribution of this component was reduced, the phase behavior of the ear canal DPOAE was not altered, suggesting that the breaking from DPOAE phase invariance at low frequencies is an outcome of apical-basal differences in cochlear mechanics. The deviation from DPOAE phase invariance appears to be a manifestation of the breaking from approximate scaling symmetry in the human cochlear apex.

Reliability and concurrent validity of TRAZER compared to three-dimensional motion capture.

BACKGROUND: Efficient neural processing of visuospatial and proprioceptive input appears to be crucial for avoidance of sport injury. As such, clinically-feasible tests are needed to identify deficiencies found by advanced neuroimaging and electrophysiological tests. Three-dimensional motion capture in a laboratory setting is currently the gold standard for measurement of human movement parameters but is costly and requires extensive training. Non-immersive virtual reality systems with body motion tracking, such as TRAZER, may provide a clinically-feasible and portable means of acquiring similar variables. Test-retest reliability and concurrent validity of these systems are currently lacking. AIM: The aim of the study was to assess the concurrent validity of the TRAZER single-camera system with 3D motion capture system and to assess the test-retest reliability of TRAZER's whole-body reactive agility metrics. METHODS: Participants - For validity, 13 healthy individuals (24.8±3.1 years, 170.0±7.7 cm, 70.0±14.2 kg); for reliability, 18 healthy individuals (23.3±2.5 years, 168.2±11.2 cm, 78.2±17.8 kg). Design - Validity was a single-session cross-sectional study. Reliability was a 3 consecutive day test-retest study. Setting-Controlled laboratory study. Intervention - Assessments utilized randomized movements in eight directions for forty total repetitions as designated by the TRAZER system. TRAZER protocol was simultaneously tracked by Vicon Motion Capture and the TRAZER system. Reliability data were captured on three consecutive days by the TRAZER system. Main Outcome Measures - Maximum acceleration, maximum velocity, and total distance were recorded for validation. In addition to these measures, maximum deceleration, average velocity, average acceleration, average deceleration, and average reaction time were collected for reliability. RESULTS: Overall, a lack of agreement exists between maximum outputs for TRAZER and 3D motion capture (velocity r=0.808, acceleration r=-0.090), but total distance correlation was high (r =.961). ICC values between days 1-2-3 for average measures were high (average velocity=0.847, average acceleration=0.919, and average deceleration=0.948) with the exception of average reaction time being fair (ICC=0.536). ICCs for maximum measures showed a much smaller correlation between days (velocity=0.654, acceleration=0.171, and deceleration=0.416). CONCLUSIONS: Even though there is a lack of strong concurrent validity between measures obtained from TRAZER and 3D motion capture systems, there is strong test-retest reliability of the TRAZER system. The applicability of these findings makes TRAZER clinically relevant in scenarios requiring pre- and post-testing for return to play decisions, or monitoring of a training regimen where demonstration of validation to a gold standard measurement is not relevant. RELEVANCE FOR PATIENTS: When test-retest capability is desired, such as in return-to-play protocols following an injury, Trazer is a reliable option.

Influence of calibration method on distortion-product otoacoustic emission measurements: II. threshold prediction.

OBJECTIVES: Distortion-product otoacoustic emission (DPOAE) stimulus calibrations are typically performed in sound pressure level (SPL) before DPOAE measurements. These calibrations may yield unpredictable DPOAE response levels, presumably because of the presence of standing waves in the ear canal. Forward pressure level (FPL) has been proposed as an alternative method for stimulus calibration because it avoids complications due to standing waves. DPOAE thresholds after four FPL calibrations and one SPL calibration were compared with behavioral thresholds to determine which calibration results in data that yield the highest correlations between the two threshold estimates. DESIGN: Fifty-two subjects with normal hearing and 103 subjects with hearing loss participated in this study, with ages ranging from 11 to 75 yr. These were the same individuals whose data were used to address the influence of calibration method on test performance in an accompanying article. DPOAE input/output (I/O) functions were obtained at f2 frequencies of 2, 3, 4, 6, and 8 kHz with the primary frequency ratio fixed at f2/f1 approximately 1.22. L(1) was set according to the equation L(1) = 0.4 L(2) + 39 with L(2) levels ranging from -20 to 70 dB SPL and FPL in 5-dB steps. I/O functions were obtained at each frequency for each of the five stimulus calibrations: SPL, daily FPL at room temperature, daily FPL at body temperature, reference FPL at room temperature, and reference FPL at body temperature. DPOAE thresholds were estimated using two methods. In the first method, DPOAE threshold was taken as the lowest L(2) for which DPOAE level is 3 dB or greater than the noise floor (signal- to-noise ratio > or =3 dB). In a second method, a linear regression method first described by Boege & Janssen (2002) and later adapted by Gorga et al. (2003), all DPOAE levels in each I/O function are converted to linear pressure and extrapolated to 0 microPa, at which the L(2) is taken as threshold. Correlations of DPOAE thresholds with behavioral thresholds were obtained for each frequency, calibration method, and threshold-prediction method. RESULTS: Correlations were greatest for frequencies of 3 to 6 kHz and lowest for 8 kHz, consistent with previous frequency effects. Calibration method made little difference in correlations between DPOAE and behavioral thresholds at any frequency. A small difference was noted in correlations for the two threshold prediction methods, with the linear regression method yielding slightly higher correlations at all frequencies. CONCLUSIONS: Little difference in threshold correlations was observed among the five calibration methods used to calibrate the stimuli before DPOAE measurements. These results were not anticipated, given the known effects of standing waves on ear-canal estimates of SPL at the plane of the probe. In addition, there was no effect of temperature (body versus room) or timing (daily versus reference) for FPL calibrations. It may be important to note that differences between SPL and FPL calibrations should not be seen if a standing wave does not occur at the plane of the probe at or near the frequency being tested. The frequencies (2 to 8 kHz) were chosen because it was expected that effects from standing waves would occur between these frequencies because of the typical lengths of ear canals for the age group tested. Because measurements were taken at only five discrete frequencies in the interval, it is possible that standing waves were present but did not affect the specific test frequencies. In total, these results suggest that SPL calibrations may be adequate when attempting to predict pure-tone thresholds from DPOAEs, despite the fact that they are known to be susceptible to errors associated with standing waves.

Influence of calibration method on distortion-product otoacoustic emission measurements: I. test performance.

OBJECTIVE: Calibration errors in distortion-product otoacoustic emission (DPOAE) measurements because of standing waves cause unpredictable changes in stimulus and DPOAE response level. The purpose of this study was to assess the extent to which these errors affect DPOAE test performance. Standard calibration procedures use sound pressure level (SPL) to determine specified levels. Forward pressure level (FPL) is an alternate calibration method that is less susceptible to standing waves. However, FPL derivation requires prior cavity measurements, which have associated variability. In an attempt to address this variability, four FPL methods were compared with SPL: a reference calibration derived from 25 measurements before all data collection and a daily calibration measurement, both of which were made at body and room temperature. DESIGN: Data were collected from 52 normal-hearing and 103 hearing-impaired subjects. DPOAEs were measured for f2 frequencies ranging from 2 to 8 kHz in half-octave steps, with L2 ranging from -20 to 70 dB SPL (5-dB steps). At each f2, DPOAEs were measured in five calibration conditions: SPL, daily FPL at body temperature (daily body), daily FPL at room temperature (daily room), reference FPL at body temperature (ref body), and reference FPL at room temperature (ref room). Data were used to construct receiver operating characteristic (ROC) curves for each f2, calibration method, and L2. From these curves, areas under the ROC curve (AROC) were estimated. RESULTS: The results of this study are summarized by the following observations: (1) DPOAE test performance was sensitive to stimulus level, regardless of calibration method, with the best test performance observed for moderate stimulus level conditions. (2) An effect of frequency was observed for all calibration methods, with the best test performance at 6 kHz and the worst performance at 8 kHz. (3) At clinically applicable stimulus levels, little difference in test performance among calibration methods was noted across frequencies, except at 8 kHz. At 8 kHz, FPL-based calibration methods provided superior performance compared with the standard SPL calibration. (4) A difference between FPL calibration methods was observed at 8 kHz, with the best test performance occurring for daily calibrations at body temperature. CONCLUSIONS: With the exception of 8 kHz, there was little difference in test performance across calibration methods. At 8 kHz, AROCs and specificities for fixed sensitivities indicate that FPL-based calibration methods provide superior performance compared with the standard SPL calibration for clinically relevant levels. Temperature may have an impact on FPL calculations relative to DPOAE test performance. Although the differences in AROC among calibration procedures were not statistically significant, the present results indicate that standing wave errors may impact DPOAE test performance and can be reduced by using FPL, although the largest effects were restricted to 8 kHz.

Impaired Geotaxis as a Novel Phenotype of Nora Virus Infection of Drosophila melanogaster.

Nora virus (NV) is a picorna-like virus that contains a positive-sense, single-stranded RNA genome. The virus infects Drosophila melanogaster with no known characterized phenotype. In this study, geotaxis assays and longevity analyses were used to determine if Nora virus infection affects D. melanogaster's locomotor ability. In addition, Drosophila C virus (DCV), a well-characterized D. melanogaster virus, was used as a positive control, as it has previously shown a locomotor defect in infected flies. Stocks infected with NV (NV+) and DCV (DCV+) and virus-free (NV-/DCV-) stocks were established. Over a 3-year period, approximately 2,500 virgin female flies were tested for geotaxis and longevity using Kaplan-Meier analyses, as well as the Cox Proportional Hazards regression for survivorship. There was a significant decrease in the geotaxis when the D. melanogaster flies were infected with Nora virus compared to uninfected controls, but no difference was found between DCV+ and NV+ trials. There were not significant differences in longevity between the three groups. This is the first time that a phenotype has been recorded in association with Nora virus infection. Overall, the data demonstrate that geotaxis dysfunction may be a phenotypic hallmark of Nora virus infection.