A role for decorin in improving motor deficits after traumatic brain injury.

Traumatic brain injury (TBI) is the leading cause of death and disability due to injury worldwide. Extracellular matrix (ECM) remodeling is known to significantly contribute to TBI pathophysiology. Glycosaminoglycans, which are long-chain, variably sulfated polysaccharides abundant within the ECM, have previously been shown to be substantially altered after TBI. In this study, we sought to delineate the dynamics of glycosaminoglycan alterations after TBI and discover the precise biologic processes responsible for observed glycosaminoglycan changes after injury. We performed state-of-the art mass spectrometry on brain tissues isolated from mice after TBI or craniotomy-alone. We observed dynamic changes in glycosaminoglycans at Day 1 and 7 post-TBI, with heparan sulfate, chondroitin sulfate, and hyaluronan remaining significantly increased after a week vis-à-vis craniotomy-alone tissues. We did not observe appreciable changes in circulating glycosaminoglycans in mice after experimental TBI compared to craniotomy-alone nor in patients with TBI and severe polytrauma compared to control patients with mild injuries, suggesting increases in injury site glycosaminoglycans are driven by local synthesis. We subsequently performed an unbiased whole genome transcriptomics analysis on mouse brain tissues 7 days post-TBI and discovered a significant induction of hyaluronan synthase 2, glypican-3, and decorin. The functional role of decorin after injury was further examined through multimodal behavioral testing comparing wild-type and Dcn-/- mice. We discovered that genetic ablation of Dcn led to an overall negative effect of TBI on function, exacerbating motor impairments after TBI. Collectively, our results provide a spatiotemporal characterization of post-TBI glycosaminoglycan alterations in the brain ECM and support an important adaptive role for decorin upregulation after TBI.

Molecular ontology of the parabrachial nucleus.

Diverse neurons in the parabrachial nucleus (PB) communicate with widespread brain regions. Despite evidence linking them to a variety of homeostatic functions, it remains difficult to determine which PB neurons influence which functions because their subpopulations intermingle extensively. An improved framework for identifying these intermingled subpopulations would help advance our understanding of neural circuit functions linked to this region. Here, we present the foundation of a developmental-genetic ontology that classifies PB neurons based on their intrinsic, molecular features. By combining transcription factor labeling with Cre fate-mapping, we find that the PB is a blend of two, developmentally distinct macropopulations of glutamatergic neurons. Neurons in the first macropopulation express Lmx1b (and, to a lesser extent, Lmx1a) and are mutually exclusive with those in a second macropopulation, which derive from precursors expressing Atoh1. This second, Atoh1-derived macropopulation includes many Foxp2-expressing neurons, but Foxp2 also identifies a subset of Lmx1b-expressing neurons in the Kölliker-Fuse nucleus (KF) and a population of GABAergic neurons ventrolateral to the PB ("caudal KF"). Immediately ventral to the PB, Phox2b-expressing glutamatergic neurons (some coexpressing Lmx1b) occupy the KF, supratrigeminal nucleus, and reticular formation. We show that this molecular framework organizes subsidiary patterns of adult gene expression (including Satb2, Calca, Grp, and Pdyn) and predicts output projections to the amygdala (Lmx1b), hypothalamus (Atoh1), and hindbrain (Phox2b/Lmx1b). Using this molecular ontology to organize, interpret, and communicate PB-related information could accelerate the translation of experimental findings from animal models to human patients.

Efferent projections of CGRP/Calca-expressing parabrachial neurons in mice.

The parabrachial nucleus (PB) is composed of glutamatergic neurons at the midbrain-hindbrain junction. These neurons form many subpopulations, one of which expresses Calca, which encodes the neuropeptide calcitonin gene-related peptide (CGRP). This Calca-expressing subpopulation has been implicated in a variety of homeostatic functions, but the overall distribution of Calca-expressing neurons in this region remains unclear. Also, while previous studies in rats and mice have identified output projections from CGRP-immunoreactive or Calca-expressing neurons, we lack a comprehensive understanding of their efferent projections. We began by identifying neurons with Calca mRNA and CGRP immunoreactivity in and around the PB, including populations in the locus coeruleus and motor trigeminal nucleus. Calca-expressing neurons in the PB prominently express the mu opioid receptor (Oprm1) and are distinct from neighboring neurons that express Foxp2 and Pdyn. Next, we used Cre-dependent anterograde tracing with synaptophysin-mCherry to map the efferent projections of these neurons. Calca-expressing PB neurons heavily target subregions of the amygdala, bed nucleus of the stria terminalis, basal forebrain, thalamic intralaminar and ventral posterior parvicellular nuclei, and hindbrain, in different patterns depending on the injection site location within the PB region. Retrograde axonal tracing revealed that the previously unreported hindbrain projections arise from a rostral-ventral subset of CGRP/Calca neurons. Finally, we show that these efferent projections of Calca-expressing neurons are distinct from those of neighboring PB neurons that express Pdyn. This information provides a detailed neuroanatomical framework for interpreting experimental work involving CGRP/Calca-expressing neurons and opioid action in the PB region.

Development and Assessment of a Microcomputed Tomography Compatible Five Degrees-of-Freedom Knee Joint Motion Simulator.

Currently available knee joint kinematic tracking systems fail to nondestructively capture the subtle variation in joint and soft tissue kinematics that occur in native, injured, and reconstructed joint states. Microcomputed tomography (CT) imaging has the potential as a noninvasive, high-resolution kinematic tracking system, but no dynamic simulators exist to take advantage of this. The purpose of this work was to develop and assess a novel micro-CT compatible knee joint simulator to quantify the knee joint's kinematic and kinetic response to clinically (e.g., pivot shift test) and functionally (e.g., gait) relevant loading. The simulator applies closed-loop, load control over four degrees-of-freedom (DOF) (internal/external rotation, varus/valgus rotation, anterior/posterior translation, and compression/distraction), and static control over a fifth degree-of-freedom (flexion/extension). Simulator accuracy (e.g., load error) and repeatability (e.g., coefficient of variation) were assessed with a cylindrical rubber tubing structure and a human cadaveric knee joint by applying clinically and functionally relevant loads along all active axes. Micro-CT images acquired of the joint at a loaded state were then used to calculate joint kinematics. The simulator loaded both the rubber tubing and the cadaveric specimen to within 0.1% of the load target, with an intertrial coefficient of variation below 0.1% for all clinically relevant loading protocols. The resultant kinematics calculated from the acquired images agreed with previously published values, and produced errors of 1.66 mm, 0.90 mm, 4.41 deg, and 1.60 deg with respect to anterior translation, compression, internal rotation, and valgus rotation, respectively. All images were free of artifacts and showed knee joint displacements in response to clinically and functionally loading with isotropic CT image voxel spacing of 0.15 mm. The results of this study demonstrate that the joint-motion simulator is capable of applying accurate, clinically and functionally relevant loads to cadaveric knee joints, concurrent with micro-CT imaging. Nondestructive tracking of bony landmarks allows for the precise calculation of joint kinematics with less error than traditional optical tracking systems.

Patient-specific cardiac phantom for clinical training and preprocedure surgical planning.

Minimally invasive mitral valve repair procedures including MitraClip® are becoming increasingly common. For cases of complex or diseased anatomy, clinicians may benefit from using a patient-specific cardiac phantom for training, surgical planning, and the validation of devices or techniques. An imaging compatible cardiac phantom was developed to simulate a MitraClip® procedure. The phantom contained a patient-specific cardiac model manufactured using tissue mimicking materials. To evaluate accuracy, the patient-specific model was imaged using computed tomography (CT), segmented, and the resulting point cloud dataset was compared using absolute distance to the original patient data. The result, when comparing the molded model point cloud to the original dataset, resulted in a maximum Euclidean distance error of 7.7 mm, an average error of 0.98 mm, and a standard deviation of 0.91 mm. The phantom was validated using a MitraClip® device to ensure anatomical features and tools are identifiable under image guidance. Patient-specific cardiac phantoms may allow for surgical complications to be accounted for preoperative planning. The information gained by clinicians involved in planning and performing the procedure should lead to shorter procedural times and better outcomes for patients.

The effects of dual-tasking on arm muscle responses in young and older adults.

This study examined whether dual-tasking affects an individual's ability to generate arm muscle responses following a loss of balance. Nineteen young and 16 older adults recovered their balance in response to a surface translation. This balance task was either completed on its own or while counting backwards by 2's (easy counting difficulty) or 7's (hard counting difficulty). With increasing counting difficulty, less attentional resources were assumed to be available for balance recovery. The ability to generate arm muscle responses was quantified through the measurement of electromyographic (EMG) onset latencies and amplitudes from three arm muscles. Results indicated that the attentional requirements of the counting task did not greatly affect EMG onset latencies or amplitudes for both young and older adults. Even when an effect was observed, the magnitude of change was small (e.g., ∼3ms earlier EMG onset and ∼2.0%MVC smaller EMG amplitude during the dual- compared to the single-task conditions). Thus, the generation of arm muscle responses do not appear to require a significant amount of attentional resources and the decreased ability to cope with cognitive interference with ageing is unlikely to explain why older adults have difficulty in generating arm responses following a loss of balance.