Phagocytosis Of Debris Research Articles

Invasion of microglia/macrophages and granulocytes into the Mauthner axon myelin sheath following spinal cord injury of the adult goldfish, Carassius auratus.

Rapid activation of resident glia occurs after spinal cord injury. Somewhat later, innate and adaptive immune responses occur with the invasion of peripheral immune cells into the wound site. The activation of resident and peripheral immune cells has been postulated to play harmful as well as beneficial roles in the regenerative process. Mauthner cells, large identifiable neurons located in the hindbrain of most fish and amphibians, provided the opportunity to study the morphological relationship between reactive cells and Mauthner axons (M-axons) severed by spinal cord crush or by selective axotomy. After crossing in the hindbrain, the M-axons of adult goldfish, Carassius auratus, extend the length of the spinal cord. Following injury, the M-axon undergoes retrograde degeneration within its myelin sheath creating an axon-free zone (proximal dieback zone). Reactive cells invade the wound site, enter the axon-free dieback zone and are observed in the vicinity of the retracted M-axon tip as early as 3 hr postinjury. Transmission electron microscopy allowed the detection of microglia/macrophages and granulocytes, some of which appear to be neutrophil-like, at each of these locations. We believe that this is the first report of the invasion of such cells within the myelin sheath of an identifiable axon in the vertebrate central nervous system (CNS). We speculate that microglia/macrophages and granulocytes that are attracted within a few hours to the damaged M-axon are part of an inflammatory response that allows phagocytosis of debris and plays a role in the regenerative process. Our results provide the baseline from which to utilize immunohistochemical and genetic approaches to elucidate the role of non-neuronal cells in the regenerative process of a single axon in the vertebrate CNS.

Promotion of microglial phagocytosis by tuftsin stimulates remyelination in experimental autoimmune encephalomyelitis.

Microglia were once thought to serve a pathogenic role in demyelinating diseases, particularly in multiple sclerosis (MS). However, it has recently been shown that in the experimental autoimmune encephalomyelitis (EAE) model of MS, microglia could serve a protective role by promoting remyelination via the efficient removal of apoptotic cells, the phagocytosis of debris and the support of myelinating oligodendrocytes. The aim of the present study was to determine if the effect of microglia could promote the recovery of EAE and attenuate symptoms in EAE. The severity of EAE was assessed by clinical scores, pathologic changes revealed by luxol fast blue staining and immunohistochemical techniques. The results suggested that microglia reduced clinical scores in mice, suppressed ongoing severe EAE and promoted remyelination and recovery in EAE mice. In addition, following induction with tuftsin, the M1/M2 cytokine balance was shifted, downregulating the proinflammatory M1 response and upregulating the anti-inflammatory M2 response. Generally, microglia can stimulate remyelination, which serves a protective role in different phases of EAE and may represent a potential therapeutic strategy for the treatment of MS.

Migratory Neural Crest Cells Phagocytose Dead Cells in the Developing Nervous System.

During neural tube closure and spinal cord development, many cells die in both the central and peripheral nervous systems (CNS and PNS, respectively). However, myeloid-derived professional phagocytes have not yet colonized the trunk region during early neurogenesis. How apoptotic cells are removed from this region during these stages remains largely unknown. Using live imaging in zebrafish, we demonstrate that neural crest cells (NCCs) respond rapidly to dying cells and phagocytose cellular debris around the neural tube. Additionally, NCCs have the ability to enter the CNSthrough motor exit point transition zones and clear debris in the spinal cord. Surprisingly, NCCs phagocytosis mechanistically resembles macrophage phagocytosis and their recruitment toward cellular debris is mediated by interleukin-1β. Taken together, our results reveal a role for NCCs in phagocytosis of debris in the developing nervous system before the presence of professional phagocytes.

Neuronal activity and microglial activation support corticospinal tract and proprioceptive afferent sprouting in spinal circuits after a corticospinal system lesion

Spared corticospinal tract (CST) and proprioceptive afferent (PA) axons sprout after injury and contribute to rewiring spinal circuits, affecting motor recovery. Loss of CST connections post-injury results in corticospinal signal loss and associated reduction in spinal activity. We investigated the role of activity loss and injury on CST and PA sprouting. To understand activity-dependence after injury, we compared CST and PA sprouting after motor cortex (MCX) inactivation, produced by chronic MCX muscimol microinfusion, with sprouting after a CST lesion produced by pyramidal tract section (PTx). Activity suppression, which does not produce a lesion, is sufficient to trigger CST axon outgrowth from the active side to cross the midline and to enter the inactivated side of the spinal cord, to the same extent as PTx. Activity loss was insufficient to drive significant CST gray matter axon elongation, an effect of PTx. Activity suppression triggered presynaptic site formation, but less than PTx. Activity loss triggered PA sprouting, as PTx. To understand injury-dependent sprouting further, we blocked microglial activation and associated inflammation after PTX by chronic minocycline administration after PTx. Minocycline inhibited myelin debris phagocytosis contralateral to PTx and abolished CST axon elongation, formation of presynaptic sites, and PA sprouting, but not CST axon outgrowth from the active side to cross the midline. Our findings suggest sprouting after injury has a strong activity dependence and that microglial activation after injury supports axonal elongation and presynaptic site formation. Combining spinal activity support and inflammation control is potentially more effective in promoting functional restoration than either alone.

How the brain eats itself

The brain is the basis of the self. We may imagine it to be a relatively unchanging structure, but recent research has shown that the brain is in fact continuously changing its microstructure, and it does so by ‘eating’ itself. The processes of eating things outside the cell, including other cells, is called phagocytosis. In the brain, phagocytosis is performed by a particular type of cell called microglia, which can ‘eat’ neurons (nerve cells) or the connections between neurons (synapses). Microglia engulf neurons and synapses during development in order to sculpt the neural circuits of the brain. Into adulthood, they continue to eat synapses, pathogens and debris in order to shape memory, stop infections and clear accumulating rubbish from the brain. However, too little synaptic pruning by microglia during development may lead to autism; whilst too much eating of synapses during adolescence may contribute to schizophrenia. Too little phagocytosis of debris and protein aggregates or too much eating of synapses and neurons may cause neurodegeneration. A whole host of problems have recently been blamed on excessive phagocytosis in the brain, including aspects of obesity, aging and even sleep deprivation. In this article we will review the evidence, outline what biochemistry can contribute and discuss how to stop the brain eating too much of itself.

Abstract A148: A hydrogel platform for the delivery of specialized pro-resolving mediators to treat chronic inflammatory disease

Abstract The resolution of acute inflammation is an active process orchestrated by mediators that prevent further recruitment of inflammatory cells and promote clearance of microbes and dying cells. Chronic inflammation can arise when resolution is diminished due to a scarcity of these pro-resolving mediators. Current immunosuppressive drugs for the treatment of chronic inflammatory diseases such as IBD, arthritis, or psoriasis temporarily block inflammation without initiating resolution or tissue repair, leading to the recurrence of chronic inflammation. Additionally, systemic administration of these drugs leaves patients susceptible to tuberculosis, lymphoma, and opportunistic infections. Resolvins are a class of evolutionarily conserved lipid mediators that return tissue to homeostasis without immunosuppression by promoting phagocytosis of debris and microbes, and preventing proinflammatory cytokine production and cellular infiltration. To enhance dose sparing and stability, we have developed a tunable hydrogel platform to deliver resolvins to various sites of chronic inflammation. For oral delivery, this hydrogel is further encapsulated into a pH-sensitive microparticle formulated using anionic polymers, to target the large intestine. Using a mouse model of colitis, we demonstrate the ability of the hydrogel to resolve intestinal inflammation, initiate repair of the intestinal barrier, and regulate the gut microbiota. Our tunable hydrogel platform could have profound effects on the prevention of gastrointestinal cancers and chronic inflammatory disease. Citation Format: Padmini Sushila Pillai, Jamie Webster, Robert Langer. A hydrogel platform for the delivery of specialized pro-resolving mediators to treat chronic inflammatory disease [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A148.

Migratory Neural Crest Cells Phagocytose Cellular Debris in the Developing Nervous System

During neural tube closure and subsequent spinal cord development, a vast number of cells undergo programmed cell death in both the central and peripheral nervous systems (CNS and PNS, respectively). However, myeloid-derived professional phagocytes have not yet colonized the trunk region during early neurogenesis. How apoptotic cells are removed from this region during early development remains largely unknown. Using live imaging in zebrafish, we demonstrate that multipotent neural crest cells (NCC) respond rapidly to dying cells and phagocytose cellular debris around the neural tube. Furthermore, migratory NCCs have the ability to enter the CNS through motor exit point transition zones and clear debris in the spinal cord. Surprisingly, NCC phagocytosis mechanistically resembles macrophage phagocytosis and their recruitment towards cellular debris is mediated by interleukin-1β signaling. Taken together, our results reveal an unexpected role of NCCs in phagocytosis of debris in the developing nervous system before the presence of professional phagocytes.

Galectin-3-Mediated Glial Crosstalk Drives Oligodendrocyte Differentiation and (Re)myelination.

Galectin-3 (Gal-3) is the only chimeric protein in the galectin family. Gal-3 structure comprises unusual tandem repeats of proline and glycine-rich short stretches bound to a carbohydrate-recognition domain (CRD). The present review summarizes Gal-3 functions in the extracellular and intracellular space, its regulation and its internalization and secretion, with a focus on the current knowledge of Gal-3 role in central nervous system (CNS) health and disease, particularly oligodendrocyte (OLG) differentiation, myelination and remyelination in experimental models of multiple sclerosis (MS). During myelination, microglia-expressed Gal-3 promotes OLG differentiation by binding glycoconjugates present only on the cell surface of OLG precursor cells (OPC). During remyelination, microglia-expressed Gal-3 favors an M2 microglial phenotype, hence fostering myelin debris phagocytosis through TREM-2b phagocytic receptor and OLG differentiation. Gal-3 is necessary for myelin integrity and function, as evidenced by myelin ultrastructural and behavioral studies from LGALS3-/- mice. Mechanistically, Gal-3 enhances actin assembly and reduces Erk 1/2 activation, leading to early OLG branching. Gal-3 later induces Akt activation and increases MBP expression, promoting gelsolin release and actin disassembly and thus regulating OLG final differentiation. Altogether, findings indicate that Gal-3 mediates the glial crosstalk driving OLG differentiation and (re)myelination and may be regarded as a target in the design of future therapies for a variety of demyelinating diseases.

In Vitro Phagocytosis of Myelin Debris by Bone Marrow-Derived Macrophages.

Bone marrow-derived macrophages (BMDMs) are mature leukocytes that serve a critical physiological role as professional phagocytes capable of clearing a variety of particles. Normally, BMDMs are restricted from the central nervous system (CNS), but following an injury, they can readily infiltrate. Once within the injured CNS tissue, BMDMs are the primary cell type responsible for the clearance of injury-derived cellular debris, including large quantities of lipid rich myelin debris. The neuropathological ramifications of BMDM infiltration and myelin debris phagocytosis within the CNS are complex and not well understood. The protocols described here, allow for the direct in vitro study of BMDMs in the context of CNS injury. We cover murine BMDM isolation and culture, myelin debris preparation, and assays to assess BMDM myelin debris phagocytosis. These techniques produce robust quantifiable results without the need for significant specialized equipment or materials, yet can be easily customized to meet the needs of researchers.

E6020, a synthetic TLR4 agonist, accelerates myelin debris clearance, Schwann cell infiltration, and remyelination in the rat spinal cord.

Oligodendrocyte progenitor cells (OPCs) are present throughout the adult brain and spinal cord and can replace oligodendrocytes lost to injury, aging, or disease. Their differentiation, however, is inhibited by myelin debris, making clearance of this debris an important step for cellular repair following demyelination. In models of peripheral nerve injury, TLR4 activation by lipopolysaccharide (LPS) promotes macrophage phagocytosis of debris. Here we tested whether the novel synthetic TLR4 agonist E6020, a Lipid A mimetic, promotes myelin debris clearance and remyelination in spinal cord white matter following lysolecithin-induced demyelination. In vitro, E6020 induced TLR4-dependent cytokine expression (TNFα, IL1β, IL-6) and NF-κB signaling, albeit at ∼10-fold reduced potency compared to LPS. Microinjection of E6020 into the intact rat spinal cord gray/white matter border induced macrophage activation, OPC proliferation, and robust oligodendrogenesis, similar to what we described previously using an intraspinal LPS microinjection model. Finally, a single co-injection of E6020 with lysolecithin into spinal cord white matter increased axon sparing, accelerated myelin debris clearance, enhanced Schwann cell infiltration into demyelinated lesions, and increased the number of remyelinated axons. In vitro assays confirmed that direct stimulation of macrophages by E6020 stimulates myelin phagocytosis. These data implicate TLR4 signaling in promoting repair after CNS demyelination, likely by stimulating phagocytic activity of macrophages, sparing axons, recruiting myelinating cells, and promoting remyelination. This work furthers our understanding of immune-myelin interactions and identifies a novel synthetic TLR4 agonist as a potential therapeutic avenue for white matter demyelinating conditions such as spinal cord injury and multiple sclerosis.

Retinoid X receptor activation reverses age-related deficiencies in myelin debris phagocytosis and remyelination.

The efficiency of central nervous system remyelination declines with age. This is in part due to an age-associated decline in the phagocytic removal of myelin debris, which contains inhibitors of oligodendrocyte progenitor cell differentiation. In this study, we show that expression of genes involved in the retinoid X receptor pathway are decreased with ageing in both myelin-phagocytosing human monocytes and mouse macrophages using a combination of in vivo and in vitro approaches. Disruption of retinoid X receptor function in young macrophages, using the antagonist HX531, mimics ageing by reducing myelin debris uptake. Macrophage-specific RXRα (Rxra) knockout mice revealed that loss of function in young mice caused delayed myelin debris uptake and slowed remyelination after experimentally-induced demyelination. Alternatively, retinoid X receptor agonists partially restored myelin debris phagocytosis in aged macrophages. The agonist bexarotene, when used in concentrations achievable in human subjects, caused a reversion of the gene expression profile in multiple sclerosis patient monocytes to a more youthful profile and enhanced myelin debris phagocytosis by patient cells. These results reveal the retinoid X receptor pathway as a positive regulator of myelin debris clearance and a key player in the age-related decline in remyelination that may be targeted by available or newly-developed therapeutics.

Abstract 2341: Characterization of macrophage behavior by 4-dimensional label free, quantitative holographic imaging

Abstract Introduction: Macrophages perform a wide range of tasks, including bacterial suppression, tumor suppression, immune-stimulation, tissue repair, angiogenesis, and cellular debris phagocytosis Behavior is controlled by a large number of chemo-stimulants, and broadly are classified into two polarization groups, M1 (classically activated, pro-inflammatory) and M2 (alternatively activated, anti-inflammatory). Understanding the macrophage phenotype is essential in the development of pharmacological agents designed to influence macrophage behavior. Due to the plethora of macrophage stimulants, it is desirable to do analysis under label-free conditions. Materials and Methods: The murine macrophage cell line, J774A.1 was analyzed untreated (M0), stimulated with LPS to drive M1, and stimulated with IL-4 to drive M2. Efficacy of the stimulations was monitored by PCR analysis of the expression levels of iNOS and TNF-Alpha (up-regulated in the M1 state) and Arginase-1 and IL-10 (up-regulated in the M2 state). Imaging data was obtained on a holographic imaging cytometer Holomonitor® M4 (Phase Holographic Imaging, Lund, Sweden). The instrument permits acquisition of time-lapse holographic image sets of unstained cells in standard vessels placed in the incubator. Images are segmented, and quantitative features are recorded for individual cells, including optical volume, thickness, area amongst others. Positional information includes X and Y positions, as well as X and Y center of mass. Data can be presented as positional track maps, position vs. feature graphs, and feature vs. feature graphs. We also obtained time-lapse videos and generated 4-dimensional (X-pos., Y-pos., cell height and time) plots in ImageJ software. Results: In untreated cells, we were able to observe the morphological and behavioral changes in cells involved in the phagocytic process. These include elongation and development of bilateral symmetry of the cells, and formation of podosomes involved in the phagocytosis of cellular remnants - all hallmarks of the M1 polarization state. We detected the same effects in cells treated with LPS under conditions known to induce M1, but without the influence of target cells. Cells present an oscillating phenotype, which we believe was caused by the redistribution of the center of mass within the cell as the polarity of the cell reverses. In Cartesian time course plots and 4D images, M1 polarized cells present linear tracking profiles. Cells treated with IL4 under conditions known to induce M2, maintained radial symmetry observable in cell images and videos. In Cartesian time course plots and 4D images, M2 polarized cells presented initial “starburst” patterns, suggesting oscillations with rotations occurring around the center of mass during cycles. Conclusions: Recently developed label-free imaging techniques provide a new tool for classifying macrophages based on cellular behavior. Citation Format: Ed Luther, Meghna Talekar, Qijun Ouyang, Mansoor Amiji. Characterization of macrophage behavior by 4-dimensional label free, quantitative holographic imaging. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2341. doi:10.1158/1538-7445.AM2015-2341

Pioglitazone treatment enhances myelin debris phagocytosis and immunoregulatory functions of monocytes from MS patients (P1.162)

Pioglitazone treatment enhances myelin debris phagocytosis and immunoregulatory functions of monocytes from MS patients (P1.162)

Complex alterations in microglial M1/M2 markers during the development of epilepsy in two mouse models.

To characterize the changes in microglial proinflammatory M1 and antiinflammatory M2 marker expression during epileptogenesis in the chronic pilocarpine and intrahippocampal kainate models. M1-activated microglia express proinflammatory cytokines driving infiltration of cells, whereas M2-activated microglia are more reparative, promoting phagocytosis of debris and expression of proteins associated with cellular stability and repair. Microglial markers were characterized as acute (3 days after status epilepticus [SE]), early chronic (21 days post-SE), and late chronic epileptic (5-12 months post-SE) time points. Following pilocarpine-SE, microglial markers were assessed by flow cytometry. Quantitative real-time polymerase chain reaction (RT-PCR) was used to measure messenger RNA (mRNA) levels of selected M1 (interleukin [IL] 1β, tumor necrosis factor α [TNFα] cluster of differentiation [CD],CD16, and CD86), interleukin-6 [IL-6], interleukin-12 [IL-12], Fc receptors 16, and CD86) and M2 (arginase 1 [Arg1], chitinase-3-like protein [Ym1], found in inflammatory zone [FIZZ-1] [FIZZ-1], mannose receptor C type-1 [CD206], interleukin-4 [IL-4], and interleukin-10 (IL-10)) markers in both models. Video-electroencephalography (EEG) recordings were used to quantify late chronic seizure frequency. Three days post-SE microglia in the pilocarpine model expressed M1 and M2 markers, but only M1 markers were upregulated after kainate-induced SE. After 3 weeks, M1/M2 marker expression was largely ablated in the hippocampal formation of both models. Small mRNA level increases of CD11b, glial fibrillary acidic protein (GFAP), and IL-1β were found in the pilocarpine model, consistent with IL-1β contributing to spontaneous seizures, whereas mRNA levels of TNFα and Ym1 were decreased. In the late chronic phase, some M1/M2 markers, IL-1β, TNFα, Arg1, Ym1, and CD206, resurged in the kainate, but not pilocarpine model, which may reflect and/or contribute to highly frequent seizures in kainate-SE mice. The common M1 upregulation acutely post-SE may signal a role early in epileptogenesis, with a more pure "inflamed" central nervous system state after kainate-SE, potentially contributing to the development of more frequent seizures. The difference may also be due to the contribution of peripheral inflammation after pilocarpine injection. In summary, the microglial inflammatory response during epileptogenesis is complex, varies between models, and appears to correlate with chronic seizure frequency.

The p53 Transcriptional Network Influences Microglia Behavior and Neuroinflammation.

The tumor-suppressor protein p53 belongs to a family of proteins that play pivotal roles in multiple cellular functions including cell proliferation, cell death, genome stability, and regulation of inflammation. Neuroinflammation is a common feature of central nervous system (CNS) pathology, and microglia are the specialized resident population of CNS myeloid cells that initiate innate immune responses. Microglia maintain CNS homeostasis through pathogen containment, phagocytosis of debris, and initiation of tissue-repair cascades. However, an unregulated pro-inflammatory response can lead to tissue injury and dysfunction in both acute and chronic inflammatory states. Therefore, regulation of the molecular signals that control the induction, magnitude, and resolution of inflammation are necessary for optimal CNS health. We and others have described a novel mechanism by which p53 transcriptional activity modulates microglia behaviors in vitro and in vivo. Activation of p53 induces expression of microRNAs (miRNAs) that support microglia pro-inflammatory functions and suppress anti-inflammatory and tissue repair behaviors. In this review, we introduce the previously described roles of the p53 signaling network and discuss novel functions of p53 in the microglia-mediated inflammatory response in CNS health and disease. Ultimately, improved understanding of the molecular regulators modulated by p53 transcriptional activity in microglia will enhance the development of rational therapeutic strategies to harness the homeostatic and tissue repair functions of microglia.

M1 and M2 Microglia Exhibit Significant Differences in their K+ Channel Expression

Microglia effector functions are widely associated with specific phenotypes, which are referred to as M1 and M2. Classically activated M1 microglia release pro-inflammatory cytokines and neurotoxic molecules and have been associated with neurological damage in ischemic stroke and Alzheimer's disease. Alternatively activated M2 microglia exhibit beneficial immunological effector functions such as phagocytosis of debris and release of anti-inflammatory and neurotrophic factors. Similar to B- and T-cells, microglia activity is regulated by calcium (Ca2+)-signaling, which is maintained by potassium (K+) channels.

Retinal dendritic cell recruitment, but not function, was inhibited in MyD88 and TRIF deficient mice.

BackgroundImmune system cells are known to affect loss of neurons due to injury or disease. Recruitment of immune cells following retinal/CNS injury has been shown to affect the health and survival of neurons in several models. We detected close, physical contact between dendritic cells and retinal ganglion cells following an optic nerve crush, and sought to understand the underlying mechanisms.MethodsCD11c-DTR/GFP mice producing a chimeric protein of diphtheria toxin receptor (DTR) and GFP from a transgenic CD11c promoter were used in conjunction with mice deficient in MyD88 and/or TRIF. Retinal ganglion cell injury was induced by an optic nerve crush, and the resulting interactions of the GFPhi cells and retinal ganglion cells were examined.ResultsRecruitment of GFPhi dendritic cells to the retina was significantly compromised in MyD88 and TRIF knockout mice. GFPhi dendritic cells played a significant role in clearing fluorescent-labeled retinal ganglion cells post-injury in the CD11c-DTR/GFP mice. In the TRIF and MyD88 deficient mice, the resting level of GFPhi dendritic cells was lower, and their influx was reduced following the optic nerve crush injury. The reduction in GFPhi dendritic cell numbers led to their replacement in the uptake of fluorescent-labeled debris by GFPlo microglia/macrophages. Depletion of GFPhi dendritic cells by treatment with diphtheria toxin also led to their displacement by GFPlo microglia/macrophages, which then assumed close contact with the injured neurons.ConclusionsThe contribution of recruited cells to the injury response was substantial, and regulated by MyD88 and TRIF. However, the presence of these adaptor proteins was not required for interaction with neurons, or the phagocytosis of debris. The data suggested a two-niche model in which resident microglia were maintained at a constant level post-optic nerve crush, while the injury-stimulated recruitment of dendritic cells and macrophages led to their transient appearance in numbers equivalent to or greater than the resident microglia.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-014-0143-1) contains supplementary material, which is available to authorized users.

Microglia toxicity in preterm brain injury

Microglia are the resident phagocytic cells of the central nervous system. During brain development they are also imperative for apoptosis of excessive neurons, synaptic pruning, phagocytosis of debris and maintaining brain homeostasis. Brain damage results in a fast and dynamic microglia reaction, which can influence the extent and distribution of subsequent neuronal dysfunction. As a consequence, microglia responses can promote tissue protection and repair following brain injury, or become detrimental for the tissue integrity and functionality. In this review, we will describe microglia responses in the human developing brain in association with injury, with particular focus on the preterm infant. We also explore microglia responses and mechanisms of microglia toxicity in animal models of preterm white matter injury and in vitro primary microglia cell culture experiments.

SCAR/WAVE-mediated processing of engulfed apoptotic corpses is essential for effective macrophage migration in Drosophila

In vitro studies have shown that SCAR/WAVE activates the Arp2/3 complex to generate actin filaments, which in many cell types are organised into lamellipodia that are thought to have an important role in cell migration. Here we demonstrate that SCAR is utilised by Drosophila macrophages to drive their developmental and inflammatory migrations and that it is regulated via the Hem/Kette/Nap1-containing SCAR/WAVE complex. SCAR is also important in protecting against bacterial pathogens and in wound repair as SCAR mutant embryos succumb more readily to both sterile and infected wounds. However, in addition to driving the formation of lamellipodia in macrophages, SCAR is required cell autonomously for the correct processing of phagocytosed apoptotic corpses by these professional phagocytes. Removal of this phagocytic burden by preventing apoptosis rescues macrophage lamellipodia formation and partially restores motility. Our results show that efficient processing of phagosomes is critical for effective macrophage migration in vivo. These findings have important implications for the resolution of macrophages from chronic wounds and the behaviour of those associated with tumours, because phagocytosis of debris may serve to prolong the presence of these cells at these sites of pathology.

Lipoic Acid Treatment after Brain Injury: Study of the Glial Reaction

After trauma brain injury, oxidative substances released to the medium provoke an enlargement of the initial lesion, increasing glial cell activation and, occasionally, an influx of immune cells into the central nervous system, developing the secondary damage. In response to these stimuli, microglia are activated to perform upregulation of intracellular enzymes and cell surface markers to propagate the immune response and phagocytosis of cellular debris. The phagocytosis of debris and dead cells is essential to limit the inflammatory reaction and potentially prevent extension of the damage to noninjured regions. Lipoic acid has been reported as a neuroprotectant by acting as an antioxidant and anti-inflammatory agent. Furthermore, angiogenic effect promoted by lipoic acid has been recently shown by our group as a crucial process for neural regeneration after brain injury. In this work, we focus our attention on the lipoic acid effect on astroglial and microglial response after brain injury.