Dorsal Root Ganglia Macrophages Maintain Osteoarthritis Pain

Pain is the major debilitating symptom of osteoarthritis (OA), which is difficult to treat. In OA patients joint tissue damage only poorly associates with pain, indicating other mechanisms contribute to OA pain. Immune cells regulate the sensory system, but little is known about the involvement of immune cells in OA pain. Here, we report that macrophages accumulate in the dorsal root ganglia (DRG) distant from the site of injury in two rodent models of OA. DRG macrophages acquired an M1-like phenotype, and depletion of DRG macrophages resolved OA pain in male and female mice. Sensory neurons innervating the damaged knee joint shape DRG macrophages into an M1-like phenotype. Persisting OA pain, accumulation of DRG macrophages, and programming of DRG macrophages into an M1-like phenotype were independent of Nav1.8 nociceptors. Inhibition of M1-like macrophages in the DRG by intrathecal injection of an IL4-IL10 fusion protein or M2-like macrophages resolved persistent OA pain. In conclusion, these findings reveal a crucial role for macrophages in maintaining OA pain independent of the joint damage and suggest a new direction to treat OA pain. SIGNIFICANCE STATEMENT: In OA patients pain poorly correlates with joint tissue changes indicating mechanisms other than only tissue damage that cause pain in OA. We identified that DRG containing the somata of sensory neurons innervating the damaged knee are infiltrated with macrophages that are shaped into an M1-like phenotype by sensory neurons. We show that these DRG macrophages actively maintain OA pain remotely and independent of joint damage. The phenotype of these macrophages is crucial for a pain-promoting role. Targeting the phenotype of DRG macrophages with either M2-like macrophages or a cytokine fusion protein that skews macrophages into an M2-like phenotype resolves OA pain. Our work reveals a mechanism that contributes to the maintenance of OA pain distant from the affected knee joint and suggests that dorsal root ganglia macrophages are a target to treat osteoarthritis chronic pain

MICALs in control of the cytoskeleton, exocytosis, and cell death

MICALs form an evolutionary conserved family of multidomain signal transduction proteins characterized by a flavoprotein monooxygenase domain. MICALs are being implicated in the regulation of an increasing number of molecular and cellular processes including cytoskeletal dynamics and intracellular trafficking. Intriguingly, some of these effects are dependent on the MICAL monooxygenase enzyme and redox signaling, while other functions rely on other parts of the MICAL protein. Recent breakthroughs in our understanding of MICAL signaling identify the ability of MICALs to bind and directly modify the actin cytoskeleton, link MICALs to the docking and fusion of exocytotic vesicles, and uncover MICALs as anti-apoptotic proteins. These discoveries could lead to therapeutic advances in neural regeneration, cancer, and other diseases

Storm-Time Relative Total Electron Content Modelling Using Machine Learning Techniques

Accurately predicting total electron content (TEC) during geomagnetic storms is still a challenging task for ionospheric models. In this work, a neural-network (NN)-based model is proposed which predicts relative TEC with respect to the preceding 27-day median TEC, during storm time for the European region (with longitudes 30°W–50°E and latitudes 32.5°N–70°N). The 27-day median TEC (referred to as median TEC), latitude, longitude, universal time, storm time, solar radio flux index F10.7, global storm index SYM-H and geomagnetic activity index Hp30 are used as inputs and the output of the network is the relative TEC. The relative TEC can be converted to the actual TEC knowing the median TEC. The median TEC is calculated at each grid point over the European region considering data from the last 27 days before the storm using global ionosphere maps (GIMs) from international GNSS service (IGS) sources. A storm event is defined when the storm time disturbance index Dst drops below 50 nanotesla. The model was trained with storm-time relative TEC data from the time period of 1998 until 2019 (2015 is excluded) and contains 365 storms. Unseen storm data from 33 storm events during 2015 and 2020 were used to test the model. The UQRG GIMs were used because of their high temporal resolution (15 min) compared to other products from different analysis centers. The NN-based model predictions show the seasonal behavior of the storms including positive and negative storm phases during winter and summer, respectively, and show a mixture of both phases during equinoxes. The model’s performance was also compared with the Neustrelitz TEC model (NTCM) and the NN-based quiet-time TEC model, both developed at the German Aerospace Agency (DLR). The storm model has a root mean squared error (RMSE) of 3.38 TEC units (TECU), which is an improvement by 1.87 TECU compared to the NTCM, where an RMSE of 5.25 TECU was found. This improvement corresponds to a performance increase by 35.6%. The storm-time model outperforms the quiet-time model by 1.34 TECU, which corresponds to a performance increase by 28.4% from 4.72 to 3.38 TECU. The quiet-time model was trained with Carrington averaged TEC and, therefore, is ideal to be used as an input instead of the GIM derived 27-day median. We found an improvement by 0.8 TECU which corresponds to a performance increase by 17% from 4.72 to 3.92 TECU for the storm-time model using the quiet-time-model predicted TEC as an input compared to solely using the quiet-time model

Semaphorin 3F Is a Bifunctional Guidance Cue for Dopaminergic Axons and Controls Their Fasciculation, Channeling, Rostral Growth, and Intracortical Targeting

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TRPC3 is a major contributor to functional heterogeneity of cerebellar Purkinje cells

espite the canonical homogeneous character of its organization, the cerebellum plays differential computational roles in distinct sensorimotor behaviors. Previously, we showed that Purkinje cell (PC) activity differs between zebrin-negative (Z–) and zebrin-positive (Z+) modules (Zhou et al., 2014). Here, using gain-of-function and loss-of-function mouse models, we show that transient receptor potential cation channel C3 (TRPC3) controls the simple spike activity of Z–, but not Z+ PCs. In addition, TRPC3 regulates complex spike rate and their interaction with simple spikes, exclusively in Z– PCs. At the behavioral level, TRPC3 loss-of-function mice show impaired eyeblink conditioning, which is related to Z– modules, whereas compensatory eye movement adaptation, linked to Z+ modules, is intact. Together, our results indicate that TRPC3 is a major contributor to the cellular heterogeneity that introduces distinct physiological properties in PCs, conjuring functional heterogeneity in cerebellar sensorimotor integration

Physicochemical characterization and evaluation of the effect of alginate-gelatin-usnic acid sponges of wound healing in rats

There is interest in the pharmaceutical area in the use of sponges, produced with biopolymers, as matrices for the controlled release of drugs. The biopolymers for pharmaceutical applications must have physicochemical and appropriate mechanical and biocompatibility. Alginate is a biopolymer, obtained from the alginic acid, which has been largely studied due to its ability to incorporate and promote the controlled release of drugs. Gelatin is a protein obtained by hydrolysis of collagen and has been used in obtaining produce hydrogels and sponges with therapeutic properties. Usnic acid (UA) is the most abundant metabolite of the lichen Cladonia substellata has proven antimicrobial, anti-inflammatory, analgesic and healing activities. In this context, the aim of this work was to physicochemical characterization and evaluate the effect of alginate-gelatin sponges, containing usnic acid on wound healing in rats. The sponges were prepared from gelatin, alginate and incorporated with usnic acid in concentrations of 1 and 5% (m/v) and freeze dried. For material characterization were used thermogravimetry/derivative thermogravimetry (TG/DTG), differential scanning calorimetry (DSC), infrared absorption spectroscopy (FTIR), scanning electron microscopy (SEM) and mechanical properties tests. The biological assay was performed with 36 rats divided into three groups: without treatment (CTR); treated with inert sponge (ALG/GEL); and treated with the sponge containing usnic acid 5% (AU 5%). After 7 and 14 days, six animals from each group were submitted to euthanasia for histological analysis. The average content of usnic acid was 60.03 and 97.38% for sponges in 1 and 5%, respectively. DSC analysis results show the absence of endothermic peak of UA pure, indicating that the drug was incorporated into the polymer system. TG/DTG curves corroborate with the DSC data and showed that the incorporation of usnic acid has not changed the thermal behavior of sponges. The FTIR spectra of sponges showed a sum corresponding to the event of usnic acid, alginate and gelatin bands, indicating the presence of these compounds in the system. The analysis by SEM showed that the incorporation of different concentrations of UA, in sponges promotes changes in microscopic structure, resulting in different mechanical behaviors. The biological assays in 7 days, showed an inflammatory reaction in the marginal and central portions of the wound, but ALG/ GEL and AU 5% groups, promoted an acceleration in the formation of granulation tissue. In 14 days, in the AU 5% group, was a clear reduction of the inflammatory component even in the central portion of the wound. Additionally, there was a marked reduction in vascular component, when compared to the CTR group with a homogeneous pattern of colagenization type III in the entire length of the scar. In conclusion, the incorporation of usnic acid has not changed the thermal stability of sponges and, in addition, the use of alginate-gelatin sponge and promotes the wound healing and the incorporation of the drug eases wound contraction and a homogeneous deposition of collagen, avoiding the formation of hipertrophic scars.Existe um interesse na área farmacêutica no uso de esponjas, produzidas com biopolímeros, como matrizes para a liberação controlada de fármacos. Os biopolímeros para aplicações farmacêuticas devem possuir propriedades físico-químicas e mecânicas adequadas, além de serem biocompatíveis. O alginato é um biopolímero, obtido a partir do ácido algínico, que tem sido largamente estudado devido a sua capacidade de incorporar compostos e promover a liberação controlada de fármacos. A gelatina é uma proteína obtida pela hidrólise do colágeno e vem sendo empregada na obtenção de hidrogéis e esponjas com propriedades terapêuticas. O ácido úsnico (AU) é o metabólito mais abundante do líquen Cladonia substellata e possui comprovada atividade antimicrobiana, anti-inflamatória, analgésica e cicatrizante. Neste contexto, o objetivo deste trabalho foi caracterizar físico-quimicamente e avaliar o efeito de esponjas de alginato-gelatina, contendo ácido úsnico sob a cicatrização em ratos. As esponjas foram preparadas a partir do alginato e gelatina, incorporadas com ácido úsnico, nas concentrações de 1 e 5% (m/v) e liofilizadas. Para caracterização do material foram utilizadas a análise termogravimétrica/termogravimétrica derivada (TG/DTG), a calorimetria exploratória diferencial (DSC), espectroscopia de absorção na região do infravermelho (FTIR), microscopia eletrônica de varredura (MEV) e testes de propriedades mecânicas. O ensaio biológico foi realizado com 36 ratos divididos em três grupos: sem tratamento (CTR); tratados com esponja inerte (ALG/GEL); e tratados com esponja contendo ácido úsnico a 5% (AU 5%). Após 7 e 14 dias, seis animais de cada grupo foram eutanasiados para análise histológica. O teor médio de ácido úsnico foi de 60,03 e 97,38% nas esponjas de 1 e 5%, respectivamente. Os resultados da análise de DSC mostram a ausência do pico endotérmico referente ao AU puro, indicando que o fármaco foi incorporado ao sistema polimérico. As curvas TG/DTG corroboram com os dados do DSC e mostraram que a incorporação do ácido úsnico não alterou o perfil geral da decomposição térmica das esponjas. Os espectros FTIR das esponjas mostraram um somatório de eventos correspondentes às bandas do ácido úsnico, alginato e gelatina, indicando a presença dos referidos compostos no sistema. A análise por MEV mostrou, que a incorporação de diferentes concentrações de AU, nas esponjas promove alterações na estrutura microscópica, resultando em diferentes comportamentos mecânicos. Os ensaios biológicos em 7 dias, mostram uma reação inflamatória nas porções centrais e marginais da ferida, porém os grupos ALG/GEL e AU 5%, promoveram uma aceleração na formação de tecido de granulação mais maduro. Em 14 dias, no grupo AU 5%, foi nítida a redução do componente inflamatório mesmo na porção central da ferida. Adicionalmente, observou-se uma marcante redução na rede vascular, quando comparado ao grupo CTR, acompanhada de um padrão homogêneo de colagenização tipo III em toda a extensão da cicatriz. Em conclusão, a incorporação do ácido úsnico não alterou a estabilidade térmica das esponjas e, além disso, o uso de esponja de alginato e gelatina favorece a cicatrização de feridas e a incorporação do fármaco facilita a contração da ferida e uma deposição homogênea de colágeno, evitando a formação de cicatrizes antiestéticas

Subdomain-mediated axon-axon signaling and chemoattraction cooperate to regulate afferent innervation of the lateral habenula

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The intracellular redox protein MICAL-1 regulates the development of hippocampal mossy fibre connections

Mical is a reduction-oxidation (redox) enzyme that functions as an unusual F-actin disassembly factor during Drosophila development. Although three Molecule interacting with CasL (MICAL) proteins exist in vertebrate species, their mechanism of action remains poorly defined and their role in vivo unknown. Here, we report that vertebrate MICAL-1 regulates the targeting of secretory vesicles containing immunoglobulin superfamily cell adhesion molecules (IgCAMs) to the neuronal growth cone membrane through its ability to control the actin cytoskeleton using redox chemistry, thereby maintaining appropriate IgCAM cell surface levels. This precise regulation of IgCAMs by MICAL-1 is essential for the lamina-specific targeting of mossy fibre axons onto CA3 pyramidal neurons in the developing mouse hippocampus in vivo. These findings reveal the first in vivo role for a vertebrate MICAL protein, expand the repertoire of cellular functions controlled through MICAL-mediated effects on the cytoskeleton, and provide insights into the poorly characterized mechanisms underlying neuronal protein cell surface expression and lamina-specific axonal targeting

The intracellular redox protein MICAL-1 regulates the development of hippocampal mossy fibre connections

Mical is a reduction-oxidation (redox) enzyme that functions as an unusual F-actin disassembly factor during Drosophila development. Although three Molecule interacting with CasL (MICAL) proteins exist in vertebrate species, their mechanism of action remains poorly defined and their role in vivo unknown. Here, we report that vertebrate MICAL-1 regulates the targeting of secretory vesicles containing immunoglobulin superfamily cell adhesion molecules (IgCAMs) to the neuronal growth cone membrane through its ability to control the actin cytoskeleton using redox chemistry, thereby maintaining appropriate IgCAM cell surface levels. This precise regulation of IgCAMs by MICAL-1 is essential for the lamina-specific targeting of mossy fibre axons onto CA3 pyramidal neurons in the developing mouse hippocampus in vivo. These findings reveal the first in vivo role for a vertebrate MICAL protein, expand the repertoire of cellular functions controlled through MICAL-mediated effects on the cytoskeleton, and provide insights into the poorly characterized mechanisms underlying neuronal protein cell surface expression and lamina-specific axonal targeting