Neurons derived from different brain regions are inherently different in vitro: a novel multiregional brain-on-a-chip.

Brain in vitro models are critically important to developing our understanding of basic nervous system cellular physiology, potential neurotoxic effects of chemicals, and specific cellular mechanisms of many disease states. In this study, we sought to address key shortcomings of current brain in vitro models: the scarcity of comparative data for cells originating from distinct brain regions and the lack of multiregional brain in vitro models. We demonstrated that rat neurons from different brain regions exhibit unique profiles regarding their cell composition, protein expression, metabolism, and electrical activity in vitro. In vivo, the brain is unique in its structural and functional organization, and the interactions and communication between different brain areas are essential components of proper brain function. This fact and the observation that neurons from different areas of the brain exhibit unique behaviors in vitro underline the importance of establishing multiregional brain in vitro models. Therefore, we here developed a multiregional brain-on-a-chip and observed a reduction of overall firing activity, as well as altered amounts of astrocytes and specific neuronal cell types compared with separately cultured neurons. Furthermore, this multiregional model was used to study the effects of phencyclidine, a drug known to induce schizophrenia-like symptoms in vivo, on individual brain areas separately while monitoring downstream effects on interconnected regions. Overall, this work provides a comparison of cells from different brain regions in vitro and introduces a multiregional brain-on-a-chip that enables the development of unique disease models incorporating essential in vivo features.NEW & NOTEWORTHY Due to the scarcity of comparative data for cells from different brain regions in vitro, we demonstrated that neurons isolated from distinct brain areas exhibit unique behaviors in vitro. Moreover, in vivo proper brain function is dependent on the connection and communication of several brain regions, underlining the importance of developing multiregional brain in vitro models. We introduced a novel brain-on-a-chip model, implementing essential in vivo features, such as different brain areas and their functional connections.

Blast-induced phenotypic switching in cerebral vasospasm.

Vasospasm of the cerebrovasculature is a common manifestation of blast-induced traumatic brain injury (bTBI) reported among combat casualties in the conflicts in Afghanistan and Iraq. Cerebral vasospasm occurs more frequently, and with earlier onset, in bTBI patients than in patients with other TBI injury modes, such as blunt force trauma. Though vasospasm is usually associated with the presence of subarachnoid hemorrhage (SAH), SAH is not required for vasospasm in bTBI, which suggests that the unique mechanics of blast injury could potentiate vasospasm onset, accounting for the increased incidence. Here, using theoretical and in vitro models, we show that a single rapid mechanical insult can induce vascular hypercontractility and remodeling, indicative of vasospasm initiation. We employed high-velocity stretching of engineered arterial lamellae to simulate the mechanical forces of a blast pulse on the vasculature. An hour after a simulated blast, injured tissues displayed altered intracellular calcium dynamics leading to hypersensitivity to contractile stimulus with endothelin-1. One day after simulated blast, tissues exhibited blast force dependent prolonged hypercontraction and vascular smooth muscle phenotype switching, indicative of remodeling. These results suggest that an acute, blast-like injury is sufficient to induce a hypercontraction-induced genetic switch that potentiates vascular remodeling, and cerebral vasospasm, in bTBI patients.

Concussion increases CA1 activity during prolonged inactivity in a familiar environment.

Although hippocampal damage plays a key role in impairments after concussion, differences in hippocampal information processing during recovery are unknown. Micro-endoscopic calcium imaging was performed before and after primary blast injury in freely behaving mice in two environments: their familiar home cage and a novel open field. Results show that after concussion CA1 activity increased in the familiar environment in which animals were awake and mostly immobile but was unaltered in a novel environment which the animals actively and constantly explored. As awake immobility parallels cognitive rest, a common treatment for patients, the results imply that prolonged cognitive rest may unwittingly impede concussion recovery.

Cardiac microphysiological devices with flexible thin-film sensors for higher-throughput drug screening.

Microphysiological systems and organs-on-chips promise to accelerate biomedical and pharmaceutical research by providing accurate in vitro replicas of human tissue. Aside from addressing the physiological accuracy of the model tissues, there is a pressing need for improving the throughput of these platforms. To do so, scalable data acquisition strategies must be introduced. To this end, we here present an instrumented 24-well plate platform for higher-throughput studies of engineered human stem cell-derived cardiac muscle tissues that recapitulate the laminar structure of the native ventricle. In each well of the platform, an embedded flexible strain gauge provides continuous and non-invasive readout of the contractile stress and beat rate of an engineered cardiac tissue. The sensors are based on micro-cracked titanium-gold thin films, which ensure that the sensors are highly compliant and robust. We demonstrate the value of the platform for toxicology and drug-testing purposes by performing 12 complete dose-response studies of cardiac and cardiotoxic drugs. Additionally, we showcase the ability to couple the cardiac tissues with endothelial barriers. In these studies, which mimic the passage of drugs through the blood vessels to the musculature of the heart, we regulate the temporal onset of cardiac drug responses by modulating endothelial barrier permeability in vitro.

Smartphone-enabled optofluidic exosome diagnostic for concussion recovery.

A major impediment to improving the treatment of concussion is our current inability to identify patients that will experience persistent problems after the injury. Recently, brain-derived exosomes, which cross the blood-brain barrier and circulate following injury, have shown great potential as a noninvasive biomarker of brain recovery. However, clinical use of exosomes has been constrained by their small size (30-100 nm) and the extensive sample preparation (>24 hr) needed for traditional exosome measurements. To address these challenges, we developed a smartphone-enabled optofluidic platform to measure brain-derived exosomes. Sample-to-answer on our chip is 1 hour, 10x faster than conventional techniques. The key innovation is an optofluidic device that can detect enzyme amplified exosome biomarkers, and is read out using a smartphone camera. Using this approach, we detected and profiled GluR2+ exosomes in the post-injury state using both in vitro and murine models of concussion.

Diagnostic tools for evaluating the impact of Focal Axonal Swellings arising in neurodegenerative diseases and/or traumatic brain injury.

BACKGROUND: Focal Axonal Swellings arise in several leading neurodegenerative diseases of the central nervous system and are hallmark features of concussions and traumatic brain injuries. Recent theories mapped how the shape of each swelling affects the propagation of spike trains and consequently the information encoded in them. Spikes can be selectively deleted, have their speed affected, or blocked depending upon the severity of the swelling. NEW METHOD: Our computational toolbox extracts meaningful geometrical parameters from sequential images of injured axon segments. The algorithm provides a principled approach for dealing with imaging distortions caused by experimental artifacts in order to extract the cross-section of an axon by detecting local symmetries, turning points and turning regions. RESULTS: Our characterization of the Focal Axonal Swelling allows for an assessment of its impact on spike propagation, leading to a color coding of the axon that highlights problematic regions for information propagation. COMPARISON WITH EXISTING METHODS: Many theoretical works reported distortions in spike propagation related to axonal enlargements. Such estimates, however, were not incorporated to a toolbox that could classify axonal swellings directly from experimental images. CONCLUSIONS: Our MATLAB toolbox thus highlights potential trouble spots of axonal morphology, and similar to car traffic maps, identify blocked or impaired routes for information flow. This computational framework is a promising starting point for diagnosing and assessing the impact of axonal swellings implicated in concussions, Alzheimer's and Parkinson's disease, Multiple Sclerosis and other neurological pathologies.

Traumatic brain injury and the neuronal microenvironment: a potential role for neuropathological mechanotransduction.

Traumatic brain injury (TBI) is linked to several pathologies for which there is a lack of understanding of disease mechanisms and therapeutic strategies. To elucidate injury mechanisms, it is important to consider how physical forces are transmitted and transduced across all spatial scales of the brain. Although the mechanical response of the brain is typically characterized by its material properties and biological structure, cellular mechanotransduction mechanisms also exist. Such mechanisms can affect physiological processes by responding to exogenous mechanical forces directed through sub-cellular components, such as extracellular matrix and cell adhesion molecules, to mechanosensitive intracellular structures that regulate mechanochemical signaling pathways. We suggest that cellular mechanotransduction may be an important mechanism underlying the initiation of cell and sub-cellular injuries ultimately responsible for the diffuse pathological damage and clinical symptoms observed in TBI, thereby providing potential therapeutic opportunities not previously explored in TBI.

Environmental consistency determines the rate of motor adaptation.

BACKGROUND: The motor system has the remarkable ability not only to learn but also to learn how fast it should learn. However, the mechanisms behind this ability are not well understood. Previous studies have posited that the rate of adaptation in a given environment is determined by Bayesian sensorimotor integration based on the amount of variability in the state of the environment. However, experimental results have failed to support several predictions of this theory. RESULTS: We show that the rate at which the motor system adapts to changes in the environment is primarily determined not by the degree to which environmental change occurs but by the degree to which the changes that do occur persist from one movement to the next, i.e., the consistency of the environment. We demonstrate a striking double dissociation whereby feedback response strength is predicted by environmental variability rather than consistency, whereas adaptation rate is predicted by environmental consistency rather than variability. We proceed to elucidate the role of stimulus repetition in speeding up adaptation and find that repetition can greatly potentiate the effect of consistency, although unlike consistency, repetition alone does not increase adaptation rate. By leveraging this understanding, we demonstrate that the rate of motor adaptation can be modulated over a range that encompasses a 20-fold increase from lowest to highest. CONCLUSIONS: Understanding the mechanisms that determine the rate of motor adaptation could lead to the principled design of improved procedures for motor training and rehabilitation. Regimens designed to control environmental consistency and repetition during training might yield faster, more robust motor learning.

A possible role for integrin signaling in diffuse axonal injury.

Over the past decade, investigators have attempted to establish the pathophysiological mechanisms by which non-penetrating injuries damage the brain. Several studies have implicated either membrane poration or ion channel dysfunction pursuant to neuronal cell death as the primary mechanism of injury. We hypothesized that traumatic stimulation of integrins may be an important etiological contributor to mild Traumatic Brain Injury. In order to study the effects of forces at the cellular level, we utilized two hierarchical, in vitro systems to mimic traumatic injury to rat cortical neurons: a high velocity stretcher and a magnetic tweezer system. In one system, we controlled focal adhesion formation in neurons cultured on a stretchable substrate loaded with an abrupt, one dimensional strain. With the second system, we used magnetic tweezers to directly simulate the abrupt injury forces endured by a focal adhesion on the neurite. Both systems revealed variations in the rate and nature of neuronal injury as a function of focal adhesion density and direct integrin stimulation without membrane poration. Pharmacological inhibition of calpains did not mitigate the injury yet the inhibition of Rho-kinase immediately after injury reduced axonal injury. These data suggest that integrin-mediated activation of Rho may be a contributor to the diffuse axonal injury reported in mild Traumatic Brain Injury.