Conditional Astrocyte Rac1KO Attenuates Hyperreflexia after Spinal Cord Injury.

Spasticity is a hyperexcitability disorder that adversely impacts functional recovery and rehabilitative efforts after spinal cord injury (SCI). The loss of evoked rate-dependent depression (RDD) of the monosynaptic H-reflex is indicative of hyperreflexia, a physiological sign of spasticity. Given the intimate relationship between astrocytes and neurons, that is, the tripartite synapse, we hypothesized that astrocytes might have a significant role in post-injury hyperreflexia and plasticity of neighboring neuronal synaptic dendritic spines. Here, we investigated the effect of selective Rac1KO in astrocytes (i.e., adult male and female mice, transgenic cre-flox system) on SCI-induced spasticity. Three weeks after a mild contusion SCI, control Rac1wt animals displayed a loss of H-reflex RDD, that is, hyperreflexia. In contrast, transgenic animals with astrocytic Rac1KO demonstrated near-normal H-reflex RDD similar to pre-injury levels. Reduced hyperreflexia in astrocytic Rac1KO animals was accompanied by a loss of thin-shaped dendritic spine density on α-motor neurons in the ventral horn. In SCI-Rac1wt animals, as expected, we observed the development of dendritic spine dysgenesis on α-motor neurons associated with spasticity. As compared with WT animals, SCI animals with astrocytic Rac1KO expressed increased levels of the glial-specific glutamate transporter, glutamate transporter-1 in the ventral spinal cord, potentially enhancing glutamate clearance from the synaptic cleft and reducing hyperreflexia in astrocytic Rac1KO animals. Taken together, our findings show for the first time that Rac1 activity in astrocytes can contribute to hyperreflexia underlying spasticity following SCI. These results reveal an opportunity to target cell-specific molecular regulators of H-reflex excitability to manage spasticity after SCI.Significance Statement Spinal cord injury leads to stretch reflex hyperexcitability, which underlies the clinical symptom of spasticity. This study shows for the first time that astrocytic Rac1 contributes to the development of hyperreflexia after SCI. Specifically, astrocytic Rac1KO reduced SCI-related H-reflex hyperexcitability, decreased dendritic spine dysgenesis on α-motor neurons, and elevated the expression of the astrocytic glutamate transporter-1 (GLT-1). Overall, this study supports a distinct role for astrocytic Rac1 signaling within the spinal reflex circuit and the development of SCI-related spasticity.

Increased astrocytic GLT-1 expression in tripartite synapses is associated with SCI-induced hyperreflexia.

Spasticity is a chronic neurological complication associated with spinal cord injury (SCI), characterized by increased muscle tone and stiffness. A physiological sign of spasticity is hyperreflexia, evident by the loss of evoked rate-dependent depression (RDD) in the H-reflex. Although previous work has shown that SCI-induced astrogliosis contributes to hyperexcitability disorders, including neuropathic pain and spasticity, it is unclear how reactive astrocytes can modulate synaptic transmission within the injured spinal cord. To study astrocytes' role in post-SCI hyperreflexia, we examined glutamate transporter-1 (GLT-1) and postsynaptic density protein 95 (PSD-95) proteins in astrocytes and neurons, respectively, within the ventral horn (lamina IX) below the level of injury (spinal segment L4-5). The close juxtaposition of GLT-1 and PSD-95 markers is a molecular correlate of tripartite synapses and is thought to be a key element in the astrocyte-induced plasticity of neuronal synapses. Our study compared animals with and without SCI-induced hyperreflexia and spasticity and investigated potential synaptic abnormalities associated with astrocyte involvement. As expected, 4 wk after SCI, we observed a loss in evoked H-reflex RDD in hindlimb electromyogram recordings, i.e., hyperreflexia, in contrast to uninjured sham. Importantly, our main findings show a significant increase in the presence of GLT-1-PSD-95 tripartite synapses in the ventral spinal cord motor regions of animals exhibiting SCI-induced hyperreflexia. Taken together, our study suggests the involvement of astrocyte-neuron synaptic complexes in the plasticity-driven progression of chronic spasticity.NEW & NOTEWORTHY The role of astrocytes in H-reflex hyperexcitability following SCI remains understudied. Our findings establish a relationship between GLT-1 expression, its proximity to neuronal PSD-95 in the spinal cord ventral horn, and the loss of H-reflex RDD, i.e., hyperreflexia. Our findings provide a new perspective on synaptic alterations and the development of SCI-related spasticity.

Kinetics of Photoreactive 4,4'-Dimethylbenzophenone Nanocrystals: Relative Contributions to Triplet Decay from Intermolecular H-Atom Transfer and Reductive Charge Transfer Quenching.

Crystals of 4,4'-dimethylbenzophenone (DMBP) are known to react by intermolecular H atom transfer, followed by radical pair recombination. To determine the contribution of the H atom transfer reaction to the deactivation of the triplet ketone, transient absorption spectra and kinetics were obtained using aqueous nanocrystalline suspensions. Single-exponential lifetimes of ca. 1185 ns with no deuterium isotope effect and inefficient product formation suggest that the reaction does not contribute significantly to the kinetics of triplet decay. By contrast, the observed lifetime is consistent with previous observations with p,p'-disubstituted benzophenones that undergo an efficient self-quenching process by a reductive charge transfer mechanism.

Dendritic Spines and Pain Memory.

Neuropathic pain is a debilitating form of pain arising from injury or disease of the nervous system that affects millions of people worldwide. Despite its prevalence, the underlying mechanisms of neuropathic pain are still not fully understood. Dendritic spines are small protrusions on the surface of neurons that play an important role in synaptic transmission. Recent studies have shown that dendritic spines reorganize in the superficial and deeper laminae of the spinal cord dorsal horn with the development of neuropathic pain in multiple models of disease or injury. Given the importance of dendritic spines in synaptic transmission, it is possible that studying dendritic spines could lead to new therapeutic approaches for managing intractable pain. In this review article, we highlight the emergent role of dendritic spines in neuropathic pain, as well as discuss the potential for studying dendritic spines for the development of new therapeutics.

Sniff-synchronized, gradient-guided olfactory search by freely moving mice.

For many organisms, searching for relevant targets such as food or mates entails active, strategic sampling of the environment. Finding odorous targets may be the most ancient search problem that motile organisms evolved to solve. While chemosensory navigation has been well characterized in microorganisms and invertebrates, spatial olfaction in vertebrates is poorly understood. We have established an olfactory search assay in which freely moving mice navigate noisy concentration gradients of airborne odor. Mice solve this task using concentration gradient cues and do not require stereo olfaction for performance. During task performance, respiration and nose movement are synchronized with tens of milliseconds precision. This synchrony is present during trials and largely absent during inter-trial intervals, suggesting that sniff-synchronized nose movement is a strategic behavioral state rather than simply a constant accompaniment to fast breathing. To reveal the spatiotemporal structure of these active sensing movements, we used machine learning methods to parse motion trajectories into elementary movement motifs. Motifs fall into two clusters, which correspond to investigation and approach states. Investigation motifs lock precisely to sniffing, such that the individual motifs preferentially occur at specific phases of the sniff cycle. The allocentric structure of investigation and approach indicates an advantage to sampling both sides of the sharpest part of the odor gradient, consistent with a serial-sniff strategy for gradient sensing. This work clarifies sensorimotor strategies for mouse olfactory search and guides ongoing work into the underlying neural mechanisms.

Probing molecular dynamics with hyperpolarized ultrafast Laplace NMR using a low-field, single-sided magnet.

Laplace NMR (LNMR) offers deep insights on diffusional and rotational motion of molecules. The so-called "ultrafast" approach, based on spatial data encoding, enables one to carry out a multidimensional LNMR experiment in a single scan, providing from 10 to 1000-fold acceleration of the experiment. Here, we demonstrate the feasibility of ultrafast diffusion-T2 relaxation correlation (D-T2) measurements with a mobile, low-field, relatively low-cost, single-sided NMR magnet. We show that the method can probe a broad range of diffusion coefficients (at least from 10-8 to 10-12 m2 s-1) and reveal multiple components of fluids in heterogeneous materials. The single-scan approach is demonstrably compatible with nuclear spin hyperpolarization techniques because the time-consuming hyperpolarization process does not need to be repeated. Using dynamic nuclear polarization (DNP), we improved the NMR sensitivity of water molecules by a factor of 105 relative to non-hyperpolarized NMR in the 0.3 T field of the single-sided magnet. This enabled us to acquire a D-T2 map in a single, 22 ms scan, despite the low field and relatively low mole fraction (0.003) of hyperpolarized water. Consequently, low-field, hyperpolarized ultrafast LNMR offers significant prospects for advanced, mobile, low-cost and high-sensitivity chemical and medical analysis.

Ultrafast Multidimensional Laplace NMR Using a Single-Sided Magnet.

Laplace NMR (LNMR) consists of relaxation and diffusion measurements providing detailed information about molecular motion and interaction. Here we demonstrate that ultrafast single- and multidimensional LNMR experiments, based on spatial encoding, are viable with low-field, single-sided magnets with an inhomogeneous magnetic field. This approach shortens the experiment time by one to two orders of magnitude relative to traditional experiments, and increases the sensitivity per unit time by a factor of three. The reduction of time required to collect multidimensional data opens significant prospects for mobile chemical analysis using NMR. Particularly tantalizing is future use of hyperpolarization to increase sensitivity by orders of magnitude, allowed by single-scan approach.

Characterization of aging and solvent treatments of painted surfaces using single-sided NMR.

Typical experiments conducted with single-sided NMR are incapable of unique chemical identification and, thus, often rely on comparative measurements in scientific study. However, cultural heritage objects have unique natures and histories, making a genuine 'control' sample a rarity and complicating many scientific investigations. In this paper, we present some comparative results enabled by such a rare, control sample. Two paintings, The Dinner and The Dance from the 1616 set Pipenpoyse Wedding, were made by the same artist with indistinguishable materials and techniques. However, despite their shared history, The Dinner has undergone varnishing and subsequent varnish removal multiple times, whereas The Dance has not. NMR measurements on these two paintings show the effect of organic-solvent-based treatments on the stiffness of the paintings as measured by T(2,eff), supporting visual and tactile observations that The Dinner is stiffer throughout its thickness than The Dance, probably due to ingress of natural resins and organic solvents into the paint and ground layers. In addition to a comparative analysis of these two paintings, initial experiments to compare solvent penetration with different varnish removal methods are described. Model canvas painting samples were treated with solvent in two ways--with free solvent on a swab and with cellulose gel thickened solvent in a tissue. Both treatment methods cause a measurable change in T(2,eff) ; however, the thickened-solvent method affects a narrower region of the model than does the free solvent.

Dense nonaqueous phase liquid architecture and dissolution in discretely fractured sandstone blocks.

Laboratory experiments were performed in discretely fractured sandstone blocks to evaluate residual dense nonaqueous phase liquid (DNAPL) architecture and dissolution. Tetrachloroethene (PCE) DNAPL residual saturations (DNAPL volume/ fracture volume) ranged between 0.18 and 0.52 for the rocks studied. DNAPL-water specific interfacial areas ranged between 19 and 57 cm2/cm3. No measurable correlation was observed between DNAPL-water interfacial area and aperture, aperture ratio, or residual saturation. DNAPL-water interfacial areas were comparable to those reported in sands with grain diameters similar to the rock apertures. However, the DNAPL residual saturation in the fractures were 2-4 times greater than in the sands, suggesting that PCE dissolution rates in rock fractures may be substantially less than in unconsolidated media, as the effective interfacial area per volume of DNAPL in rock fractures was less than in sands. Comparison of dissolution mass transfer coefficients in the bedrock fractures to corresponding mass transfer coefficients measured in sands indicated that dissolution rates in bedrock fractures were substantially less than dissolution rates measured in sands, even after normalization to DNAPL-water interfacial area. The presence of preferential water and DNAPL flow paths within the discrete fractures was shown to have a significant impact on observed DNAPL dissolution rates. DNAPL dissolution was reasonably described by a Reynolds number correlation that incorporated flow characteristics and the DNAPL-water interfacial area.

Birthplace effects on the development of female athletic talent.

This study examined the extent to which an athlete's place of birth can influence the likelihood of playing professional sport. Information regarding the birthplace of all American female athletes in the Ladies Professional Golf Association and Women's United Soccer Association was gathered from official league websites. Monte Carlo simulations were used to determine if the birthplace of these professional athletes differed in any systematic way from official census population distributions. Odds-ratios were determined for cities within specific population ranges to ascertain if the likelihood of playing professional sport was influenced in any systematic way by city size. The analyses revealed that female professional soccer players born in cities of less than 1,000,000 were over-represented, as were female professional golfers born in cities of less than 250,000. Results are consistent with those of male professional athletes in suggesting that areas of lower population provide conditions more conducive to the development of expertise than do larger city environments.