A New Role for TIMP-1 in Modulating Neurite Outgrowth and Morphology of Cortical Neurons

BACKGROUND:Tissue inhibitor of metalloproteinases-1 (TIMP-1) displays pleiotropic activities, both dependent and independent of its inhibitory activity on matrix metalloproteinases (MMPs). In the central nervous system (CNS), TIMP-1 is strongly upregulated in reactive astrocytes and cortical neurons following excitotoxic/inflammatory stimuli, but no information exists on its effects on growth and morphology of cortical neurons. PRINCIPAL FINDINGS:We found that 24 h incubation with recombinant TIMP-1 induced a 35% reduction in neurite length and significantly increased growth cones size and the number of F-actin rich microprocesses. TIMP-1 mediated reduction in neurite length affected both dendrites and axons after 48 h treatment. The effects on neurite length and morphology were not elicited by a mutated form of TIMP-1 inactive against MMP-1, -2 and -3, and still inhibitory for MMP-9, but were mimicked by a broad spectrum MMP inhibitor. MMP-9 was poorly expressed in developing cortical neurons, unlike MMP-2 which was present in growth cones and whose selective inhibition caused neurite length reductions similar to those induced by TIMP-1. Moreover, TIMP-1 mediated changes in cytoskeleton reorganisation were not accompanied by modifications in the expression levels of actin, betaIII-tubulin, or microtubule assembly regulatory protein MAP2c. Transfection-mediated overexpression of TIMP-1 dramatically reduced neuritic arbour extension in the absence of detectable levels of released extracellular TIMP-1. CONCLUSIONS:Altogether, TIMP-1 emerges as a modulator of neuronal outgrowth and morphology in a paracrine and autrocrine manner through the inhibition, at least in part, of MMP-2 and not MMP-9. These findings may help us understand the role of the MMP/TIMP system in post-lesion pre-scarring conditions

Resuscitation of Newborn Piglets. Short-Term Influence of FiO2 on Matrix Metalloproteinases, Caspase-3 and BDNF

Perinatal hypoxia-ischemia is a major cause of mortality and cerebral morbidity, and using oxygen during newborn resuscitation may further harm the brain. The aim was to examine how supplementary oxygen used for newborn resuscitation would influence early brain tissue injury, cell death and repair processes and the regulation of genes related to apoptosis, neurodegeneration and neuroprotection.Anesthetized newborn piglets were subjected to global hypoxia and then randomly assigned to resuscitation with 21%, 40% or 100% O(2) for 30 min and followed for 9 h. An additional group received 100% O(2) for 30 min without preceding hypoxia. The left hemisphere was used for histopathology and immunohistochemistry and the right hemisphere was used for in situ zymography in the corpus striatum; gene expression and the activity of various relevant biofactors were measured in the frontal cortex. There was an increase in the net matrix metalloproteinase gelatinolytic activity in the corpus striatum from piglets resuscitated with 100% oxygen vs. 21%. Hematoxylin-eosin (HE) staining revealed no significant changes. Nine hours after oxygen-assisted resuscitation, caspase-3 expression and activity was increased by 30-40% in the 100% O(2) group (n = 9/10) vs. the 21% O(2) group (n = 10; p<0.04), whereas brain-derived neurotrophic factor (BDNF) activity was decreased by 65% p<0.03.The use of 100% oxygen for resuscitation resulted in increased potentially harmful proteolytic activities and attenuated BDNF activity when compared with 21%. Although there were no significant changes in short term cell loss, hyperoxia seems to cause an early imbalance between neuroprotective and neurotoxic mechanisms that might compromise the final pathological outcome

Differential spatio-temporal regulation of MMPs in the 5xFAD mouse model of Alzheimer's disease: evidence for a pro-amyloidogenic role of MT1-MMP

International audienceMatrix metalloproteinases (MMPs) are pleiotropic endopeptidases involved in a variety of neurodegenerative/neuroinflammatory processes through their interactions with a large number of substrates. Among those, the amyloid precursor protein (APP) and the beta amyloid peptide (Aβ) are largely associated with the development of Alzheimer's disease (AD). However, the regulation and potential contribution of MMPs to AD remains unclear. In this study, we investigated the evolution of the expression of MMP-2, MMP-9, and membrane-type 1-MMP (MT1-MMP) in the hippocampus at different stages of the pathology (asymptomatic, prodromal-like and symptomatic) in the 5xFAD transgenic mouse AD model. In parallel we also followed the expression of functionally associated factors. Overall, the expression of MMP-2, MMP-9, and MT1-MMP was upregulated concomitantly with the tissue inhibitor of MMPs-1 (TIMP-1) and several markers of inflammatory/glial response. The three MMPs exhibited age- and cell-dependent upregulation of their expression, with MMP-2 and MMP-9 being primarily located to astrocytes, and MT1-MMP to neurons. MMP-9 and MT1-MMP were also prominently present in amyloid plaques. The levels of active MT1-MMP were highly upregulated in membrane-enriched fractions of hippocampus at 6 months of age (symptomatic phase), when the levels of APP, its metabolites APP C-terminal fragments (CTFs), and Aβ trimers were the highest. Overexpression of MT1-MMP in HEK cells carrying the human APP Swedish mutation (HEKswe) strongly increased β-secretase derived C-terminal APP fragment (C99) and Aβ levels, whereas MMP-2 overexpression nearly abolished Aβ production without affecting C99. Our data consolidate the emerging idea of a regulatory interplay between MMPs and the APP/Aβ system, and demonstrate for the first time the pro-amyloidogenic features of MT1-MMP. Further investigation will be justified to evaluate this MMP as a novel potential therapeutic target in AD

Vegetation, Climate and Habitability in the Marseille Basin (SE France) circa 1 Ma

International audienceThe environment of the Marseille basin in the Early Pleistocene was reconstructed through a multiproxy study of fluvial tufa deposits. Palaeomagnetic measurements revealed the Jaramillo subchron and dated the tufa to within the 0.8-1.5 Ma interval, probably between 0.9 and1.2 Ma. Sedimentological studies show varied depositional environments comprising natural dams formed by accumulations of plants promoting the development of upstream water bodies. The very negative δ 13 C values indicate that the Marseille tufa is not travertine sensu stricto but tufa deposited by local cold-water rivers. Palynological analyses indicate a semi-forested, diverse, mosaic vegetation landscape dominated by a Mediterranean pine and oak forest. Along the streams, the riparian forest was diverse and included Juglans, Castanea, Platanus and Vitis. The potential diet reconstructed from pollen was varied. The most surprising discovery was the presence of proto-cereals, which could potentially enrich the diet with carbohydrates. The identification of spores of coprophilous fungi seems to indicate the presence in situ of large herbivore herds. It is possible that, as in Anatolia, the disturbance of ecosystems by large herbivores was responsible for the genetic mutation of Poaceae and the appearance of proto-cereals. Climatic reconstructions indicate a slightly cooler and wetter climate than the present

MT5-MMP at the crossroads of neuroinflammation and amyloidogenesis: study in murine and human neural cells

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Role of Matrix Metalloproteinases in Migration and Neurotrophic Properties of Nasal Olfactory Stem and Ensheathing Cells

International audienceAdult olfactory ectomesenchymal stem cells (OE-MSCs) and olfactory ensheathing cells (OECs), both from the nasal olfactory lamina propria, display robust regenerative properties when transplanted into the nervous system, but the mechanisms supporting such therapeutic effects remain unknown. Matrix metalloproteinases (MMPs) are an important family of proteinases contributing to cell motility and axonal outgrowth across the extracellular matrix (ECM) in physiological and pathological conditions. In this study, we have characterized for the first time in nasal human OE-MSCs the expression profile of some MMPs currently associated with cell migration and invasiveness. We demonstrate different patterns of expression for MMP-1, MMP-2, MMP-9, and MT1-MMP upon cell migration when compared with nonmigrating cells. Our results establish a correspondence between the localization of these proteinases in the migration front with the ability of cells to migrate. Using various modulators of MMP activity, we also show that at least MMP-2, MMP-9, and MT1-MMP contribute to OE-MSC migration in an in vitro 3D test. Furthermore, we demonstrate under the same conditions of culture used for in vivo transplantation that OE-MSCs and OECs secrete neurotrophic factors that promote neurite outgrowth of cortical and dorsal root ganglia (DRG) neurons, as well as axo-dendritic differentiation of cortical neurons. These effects were abolished by the depletion of MMP-2 and MMP-9 from the culture conditioned media. Altogether, our results provide the first evidence that MMPs may contribute to the therapeutic features of OE-MSCs and OECs through the control of their motility and/or their neurotrophic properties. Our data provide new insight into the mechanisms of neuroregeneration and will contribute to optimization of cell therapy strategies

<i>In situ z</i>ymography in the corpus striatum.

<p>Net gelatinolytic activity increases in the striatum after hypoxia-resuscitation. Fluorescence photomicrographs of striatum sections showing <i>in situ</i> zymography in sham operated (Ctl) and hyperoxia (Hyp) controls and after reoxygenation with 21%, 40% or 100% O₂. Overall, fluorescence signal representing proteolytic activity (green) increases after hypoxia-resuscitation in the entire tissue, but the most prominent changes occur in discrete neuronal populations (arrows) in a dose-response manner. Hoechst stain was used as a nuclear marker (blue). Scale bar: 150 µm. The graph represents the quantification of net gelatinolytic activity (in arbitrary units (AU) of fluorescence) for 21% (n = 8) 50.64 (10.1), 40% (n = 8) 55.29 (5.4), 100% (n = 9) 59.51 (11.1), hyperoxia (n = 6) 35.42 (8.0) and controls (n = 6) 35.67 (6.1). There was a significant increase in net gelatinolytic activity in the corpus striatum in all groups exposed to hypoxia-reoxygenation vs the control group (p = 0.024, p = 0.002 and p<0.001 for the 21%, 40% and 100% group). The hyperoxia-group was similar to the control group. Using <i>post hoc</i> multiple comparisons between group means (Fisher LSD), we found a significant increase in the 100% oxygen group compared to the 21% oxygen group (p = 0.043). Values are expressed as a mean (±SD), *p<0.05, **p<0.01.</p

Grading of damage in striatum, hippocampus, cortex and cerebellum (HE stained sections).

<p>Grading of damage in striatum, hippocampus, cortex and cerebellum (HE stained sections).</p

Background data.

<p><i>Characterization of the study cohort before, directly after asphyxia and after reoxygenation. Values are presented as mean (±SD). Italic values show the control- and hyperoxia group at corresponding time points.</i></p

MMP-9 mRNA expression in the cortex.

<p>Relative mRNA MMP-9 expression was significantly increased in the 21% oxygen group vs. all the others: 21% (n = 10) 23.1 (19.3), 40% (n = 12) 7.8 (15.1), 100% (n = 10) 4.3 (4.2), hyperoxia (n = 11)1.8 (1.6), control (n = 6) 3.9 (4) with p = 0.029, 0.007, 0.001 and 0.017, respectively.</p