Your browser doesn't support javascript.
loading
: 20 | 50 | 100
1 - 11 de 11
2.
bioRxiv ; 2023 Aug 30.
Article En | MEDLINE | ID: mdl-37693409

Parkinson's disease (PD) is characterized by the aggregation of α-synuclein into Lewy bodies and Lewy neurites in the brain. Microglia-driven neuroinflammation may contribute to neuronal death in PD, however the exact role of microglia remains unclear and has been understudied. The A53T mutation in the gene coding for α-synuclein has been linked to early-onset PD, and exposure to A53T-mutant human α-synuclein increases the potential for inflammation of murine microglia. To date, its effect has not been studied in human microglia. Here, we used 2-dimensional cultures of human iPSC-derived microglia and transplantation of these cells into the mouse brain to assess the effects of the A53T mutation on human microglia. We found that A53T-mutant human microglia had an intrinsically increased propensity towards pro-inflammatory activation upon inflammatory stimulus. Additionally, A53T mutant microglia showed a strong decrease in catalase expression in non-inflammatory conditions, and increased oxidative stress. Our results indicate that A53T mutant human microglia display cell-autonomous phenotypes that may worsen neuronal damage in early-onset PD.

3.
Biol Psychiatry ; 93(1): 71-81, 2023 01 01.
Article En | MEDLINE | ID: mdl-36372569

BACKGROUND: Fragile X syndrome (FXS) is characterized by physical abnormalities, anxiety, intellectual disability, hyperactivity, autistic behaviors, and seizures. Abnormal neuronal development in FXS is poorly understood. Data on patients with FXS remain scarce, and FXS animal models have failed to yield successful therapies. In vitro models do not fully recapitulate the morphology and function of human neurons. METHODS: To mimic human neuron development in vivo, we coinjected neural precursor cells derived from FXS patient-derived induced pluripotent stem cells and neural precursor cells derived from corrected isogenic control induced pluripotent stem cells into the brain of neonatal immune-deprived mice. RESULTS: The transplanted cells populated the brain and a proportion differentiated into neurons and glial cells. Immunofluorescence and single and bulk RNA sequencing analyses showed accelerated maturation of FXS neurons after an initial delay. Additionally, we found increased percentages of Arc- and Egr-1-positive FXS neurons and wider dendritic protrusions of mature FXS striatal medium spiny neurons. CONCLUSIONS: This transplantation approach provides new insights into the alterations of neuronal development in FXS by facilitating physiological development of cells in a 3-dimensional context.


Fragile X Syndrome , Neural Stem Cells , Humans , Mice , Animals , Fragile X Syndrome/genetics , Fragile X Mental Retardation Protein/genetics , Fragile X Mental Retardation Protein/metabolism , Neural Stem Cells/metabolism , Neurons/metabolism , Phenotype , Brain/metabolism , Mice, Knockout
4.
Comp Med ; 72(4): 220-229, 2022 08 01.
Article En | MEDLINE | ID: mdl-35882504

Klebsiella pneumoniae (Kp) is a gram-negative opportunistic pathogen that causes severe pneumonia, pyelonephritis, and sepsis in immunocompromised hosts. During a 4-mo interval, several NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) breeders and pups in our facilities were diagnosed with Kp infections. An initial 6 adult and 1 juvenile NSG mice were submitted for necropsy and histologic examination because of acute onset of diarrhea and death. The evaluation revealed typhlocolitis in 2 of the mice and tritrichomoniasis in all 7. Escherichia coli positive for polyketide synthase (pks+) and Kp were isolated from the intestines. Given a history of sepsis due to pks+ E. coli in NSG mice in our facilities and determination of its antimicrobial susceptibility, trimethoprim-sulfamethoxazole (TMP-SMX) was administered to the colony in the drinking water for 4 wk. After this intervention, an additional 21 mice became ill or died; 11 of these mice had suppurative pneumonia, meningoencephalitis, hepatitis, metritis, pyelonephritis, or sepsis. Kp was cultured from pulmonary abscesses or blood of 10 of the mice. Whole-genome sequencing (WGS) indicated that the Kp isolates contained genes associated with phenotypes found in pore-forming Kp isolates cultured from humans with ulcerative colitis and primary sclerosing cholangitis. None of the Kp isolates exhibited a hyperviscous phenotype, but 13 of 14 were resistant to TMP-SMX. Antimicrobial susceptibility testing indicated sensitivity of the Kp to enrofloxacin, which was administered in the drinking water. Antibiotic sensitivity profiles were confirmed by WGS of the Kp strains; key virulence and resistance genes to quaternary ammonia compounds were also identified. Enrofloxacin treatment resulted in a marked reduction in mortality, and the study using the NSG mice was completed successfully. Our findings implicate intestinal translocation of Kp as the cause of pneumonia and systemic infections in NSG mice and highlight the importance of identification of enteric microbial pathogens and targeted antibiotic selection when treating bacterial infections in immunocompromised mice.


Drinking Water , Pneumonia , Pyelonephritis , Sepsis , Adult , Animals , Anti-Bacterial Agents/pharmacology , Enrofloxacin , Escherichia coli , Humans , Immunocompromised Host , Klebsiella pneumoniae , Mice , Mice, Inbred NOD , Microbial Sensitivity Tests , Trimethoprim, Sulfamethoxazole Drug Combination
5.
Cell ; 172(5): 979-992.e6, 2018 02 22.
Article En | MEDLINE | ID: mdl-29456084

Fragile X syndrome (FXS), the most common genetic form of intellectual disability in males, is caused by silencing of the FMR1 gene associated with hypermethylation of the CGG expansion mutation in the 5' UTR of FMR1 in FXS patients. Here, we applied recently developed DNA methylation editing tools to reverse this hypermethylation event. Targeted demethylation of the CGG expansion by dCas9-Tet1/single guide RNA (sgRNA) switched the heterochromatin status of the upstream FMR1 promoter to an active chromatin state, restoring a persistent expression of FMR1 in FXS iPSCs. Neurons derived from methylation-edited FXS iPSCs rescued the electrophysiological abnormalities and restored a wild-type phenotype upon the mutant neurons. FMR1 expression in edited neurons was maintained in vivo after engrafting into the mouse brain. Finally, demethylation of the CGG repeats in post-mitotic FXS neurons also reactivated FMR1. Our data establish that demethylation of the CGG expansion is sufficient for FMR1 reactivation, suggesting potential therapeutic strategies for FXS.


DNA Methylation/genetics , Fragile X Mental Retardation Protein/genetics , Fragile X Syndrome/genetics , Gene Editing , Neurons/pathology , Animals , CRISPR-Associated Protein 9/metabolism , Epigenesis, Genetic , HEK293 Cells , Heterochromatin/metabolism , Humans , Induced Pluripotent Stem Cells/metabolism , Kinetics , Male , Mice , Neurons/metabolism , Phenotype , Promoter Regions, Genetic , RNA, Guide, Kinetoplastida/metabolism , Trinucleotide Repeat Expansion/genetics
6.
Neurogenesis (Austin) ; 4(1): e1286424, 2017.
Article En | MEDLINE | ID: mdl-28265586

Post-natal hippocampal neurogenesis plays a role in hippocampal function, and neurons born post-natally participate to spatial memory and mood control. However, a great proportion of granule neurons generated in the post-natal hippocampus are eliminated during the first 3 weeks of their maturation, a mechanism that depends on their synaptic integration. In a recent study, we examined the possibility of enhancing the synaptic integration of neurons born post-natally, by specifically overexpressing synaptic cell adhesion molecules in these cells. Synaptic cell adhesion molecules are transmembrane proteins mediating the physical connection between pre- and post-synaptic neurons at the synapse, and their overexpression enhances synapse formation. Accordingly, we found that overexpressing synaptic adhesion molecules increased the synaptic integration and survival of newborn neurons. Surprisingly, the synaptic adhesion molecule with the strongest effect on new neurons' survival, Neuroligin-2A, decreased memory performances in a water maze task. We present here hypotheses explaining these surprising results, in the light of the current knowledge of the mechanisms of synaptic integration of new neurons in the post-natal hippocampus.

7.
Cereb Cortex ; 27(8): 4048-4059, 2017 08 01.
Article En | MEDLINE | ID: mdl-27473321

Postnatal hippocampal neurogenesis induces network remodeling and may participate to mechanisms of learning. In turn, the maturation and survival of newborn neurons is regulated by their activity. Here, we tested the effect of a cell-autonomous overexpression of synaptic adhesion molecules on the maturation and survival of neurons born postnatally and on hippocampal-dependent memory performances. Families of adhesion molecules are known to induce pre- and post-synaptic assembly. Using viral targeting, we overexpressed three different synaptic adhesion molecules, SynCAM1, Neuroligin-1B and Neuroligin-2A in newborn neurons in the dentate gyrus of 7- to 9-week-old mice. We found that SynCAM1 increased the morphological maturation of dendritic spines and mossy fiber terminals while Neuroligin-1B increased spine density. In contrast, Neuroligin-2A increased both spine density and size as well as GABAergic innervation and resulted in a drastic increase of neuronal survival. Surprisingly, despite increased neurogenesis, mice overexpressing Neuroligin-2A in new neurons showed decreased memory performances in a Morris water maze task. These results indicate that the cell-autonomous overexpression of synaptic adhesion molecules can enhance different aspects of synapse formation on new neurons and increase their survival. Furthermore, they suggest that the mechanisms by which new neurons integrate in the postnatal hippocampus conditions their functional implication in learning and memory.


Cell Adhesion Molecule-1/metabolism , Cell Adhesion Molecules, Neuronal/metabolism , Dentate Gyrus/metabolism , Nerve Tissue Proteins/metabolism , Neurons/metabolism , Spatial Memory/physiology , Animals , Cell Adhesion Molecule-1/genetics , Cell Adhesion Molecules, Neuronal/genetics , Cell Survival/physiology , Dentate Gyrus/cytology , Glutamic Acid/metabolism , HEK293 Cells , Humans , Male , Maze Learning/physiology , Mice, Inbred C57BL , Nerve Tissue Proteins/genetics , Neurogenesis/physiology , Neuronal Plasticity/physiology , Neurons/cytology , Neuropsychological Tests , Synapses/metabolism , gamma-Aminobutyric Acid/metabolism
8.
Sci Rep ; 6: 39310, 2016 12 19.
Article En | MEDLINE | ID: mdl-27991559

Neurexins are transmembrane synaptic cell adhesion molecules involved in the development and maturation of neuronal synapses. In the present study, we report that Nrxn3ß is processed by the metalloproteases ADAM10, ADAM17, and by the intramembrane-cleaving protease γ-secretase, producing secreted neurexin3ß (sNrxn3ß) and a single intracellular domain (Nrxn3ß-ICD). We further completed the full characterization of the sites at which Nrxn3ß is processed by these proteases. Supporting the physiological relevance of the Nrxn3ß processing, we demonstrate in vivo a significant effect of the secreted shedding product sNrxn3ß on the morphological development of adult newborn neurons in the mouse hippocampus. We show that sNrxn3ß produced by the cells of the dentate gyrus increases the spine density of newborn neurons whereas sNrxn3ß produced by the newborn neuron itself affects the number of its mossy fiber terminal extensions. These results support a pivotal role of sNrxn3ß in plasticity and network remodeling during neuronal development.


ADAM10 Protein/metabolism , Amyloid Precursor Protein Secretases/metabolism , Hippocampus/cytology , Membrane Proteins/metabolism , Nerve Tissue Proteins/metabolism , Neurons/drug effects , Neurons/physiology , Protein Processing, Post-Translational , ADAM17 Protein/metabolism , Animals , Animals, Newborn , Mice, Inbred C57BL , Proteolysis
9.
Brain Struct Funct ; 220(4): 2027-42, 2015 Jul.
Article En | MEDLINE | ID: mdl-24748560

The adult dentate gyrus produces new neurons that morphologically and functionally integrate into the hippocampal network. In the adult brain, most excitatory synapses are ensheathed by astrocytic perisynaptic processes that regulate synaptic structure and function. However, these processes are formed during embryonic or early postnatal development and it is unknown whether astrocytes can also ensheathe synapses of neurons born during adulthood and, if so, whether they play a role in their synaptic transmission. Here, we used a combination of serial-section immuno-electron microscopy, confocal microscopy, and electrophysiology to examine the formation of perisynaptic processes on adult-born neurons. We found that the afferent and efferent synapses of newborn neurons are ensheathed by astrocytic processes, irrespective of the age of the neurons or the size of their synapses. The quantification of gliogenesis and the distribution of astrocytic processes on synapses formed by adult-born neurons suggest that the majority of these processes are recruited from pre-existing astrocytes. Furthermore, the inhibition of astrocytic glutamate re-uptake significantly reduced postsynaptic currents and increased paired-pulse facilitation in adult-born neurons, suggesting that perisynaptic processes modulate synaptic transmission on these cells. Finally, some processes were found intercalated between newly formed dendritic spines and potential presynaptic partners, suggesting that they may also play a structural role in the connectivity of new spines. Together, these results indicate that pre-existing astrocytes remodel their processes to ensheathe synapses of adult-born neurons and participate to the functional and structural integration of these cells into the hippocampal network.


Astrocytes/physiology , Hippocampus/cytology , Neurons/cytology , Aldehyde Dehydrogenase 1 Family , Animals , Astrocytes/ultrastructure , Bromodeoxyuridine/metabolism , Dendritic Spines/drug effects , Dendritic Spines/metabolism , Excitatory Postsynaptic Potentials/drug effects , Excitatory Postsynaptic Potentials/genetics , Gene Expression Regulation/genetics , Glial Fibrillary Acidic Protein/genetics , Glial Fibrillary Acidic Protein/metabolism , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Isoenzymes/genetics , Isoenzymes/metabolism , Kainic Acid/analogs & derivatives , Kainic Acid/pharmacology , Mice , Mice, Inbred C57BL , Mice, Transgenic , Microscopy, Confocal , Microscopy, Immunoelectron , Neurogenesis/drug effects , Neurogenesis/genetics , Neurons/drug effects , Patch-Clamp Techniques , Phosphopyruvate Hydratase/metabolism , Retinal Dehydrogenase/genetics , Retinal Dehydrogenase/metabolism , S100 Calcium Binding Protein beta Subunit/metabolism , Synapses/physiology , Synapses/ultrastructure , Synaptic Transmission/drug effects , Synaptic Transmission/genetics
10.
Anesthesiology ; 118(3): 602-10, 2013 Mar.
Article En | MEDLINE | ID: mdl-23314165

BACKGROUND: Adult neurogenesis occurs in the hippocampus of most mammals, including humans, and plays an important role in hippocampal-dependent learning. This process is highly regulated by neuronal activity and might therefore be vulnerable to anesthesia. In this article, the authors investigated this possibility by evaluating the impact of propofol anesthesia on mouse hippocampal neurons generated during adulthood, at two functionally distinct maturational stages of their development. METHODS: Adult-born hippocampal neurons were identified using the cell proliferation marker bromodeoxyuridine or a retroviral vector expressing the green fluorescent protein in dividing cells and their progenies. Eleven or 17 days after the labeling procedure, animals (n = 3-5 animals per group) underwent a 6-h-long propofol anesthesia. Twenty-one days after labeling, the authors analyzed the survival, differentiation, and morphologic maturation of adult-born neurons using confocal microscopy. RESULTS: Propofol impaired the survival and maturation of adult-born neurons in an age-dependent manner. Anesthesia induced a significant decrease in the survival of neurons that were 17 days old at the time of anesthesia, but not of neurons that were 11 days old. Similarly, propofol anesthesia significantly reduced the dendritic maturation of neurons generated 17 days before anesthesia, without interfering with the maturation of neurons generated 11 days before anesthesia. CONCLUSIONS: These results reveal that propofol impairs the survival and maturation of adult-born hippocampal neurons in a developmental stage-dependent manner in mice.


Anesthesia/adverse effects , Cellular Senescence/drug effects , Hippocampus/drug effects , Hippocampus/pathology , Neurogenesis/drug effects , Neurons/drug effects , Neurons/pathology , Propofol/toxicity , Animals , Cell Survival/drug effects , Male , Mice , Mice, Inbred C57BL
11.
J Cell Mol Med ; 16(6): 1206-22, 2012 Jun.
Article En | MEDLINE | ID: mdl-21762376

According to epidemiological studies, type-2 diabetes increases the risk of Alzheimer's disease. Here, we induced hyperglycaemia in mice overexpressing mutant amyloid precursor protein and presenilin-1 (APdE9) either by cross-breeding them with pancreatic insulin-like growth factor 2 (IGF-2) overexpressing mice or by feeding them with high-fat diet. Glucose and insulin tolerance tests revealed significant hyperglycaemia in mice overexpressing IGF-2, which was exacerbated by high-fat diet. However, sustained hyperinsulinaemia and insulin resistance were observed only in mice co-expressing IGF-2 and APdE9 without correlation to insulin levels in brain. In behavioural tests in aged mice, APdE9 was associated with poor spatial learning and the combination of IGF-2 and high-fat diet further impaired learning. Neither high-fat diet nor IGF-2 increased ß-amyloid burden in the brain. In male mice, IGF-2 increased ß-amyloid 42/40 ratio, which correlated with poor spatial learning. In contrast, inhibitory phosphorylation of glycogen synthase kinase 3ß, which correlated with good spatial learning, was increased in APdE9 and IGF-2 female mice on standard diet, but not on high-fat diet. Interestingly, high-fat diet altered τ isoform expression and increased phosphorylation of τ at Ser202 site in female mice regardless of genotype. These findings provide evidence for new regulatory mechanisms that link type-2 diabetes and Alzheimer pathology.


Alzheimer Disease/genetics , Diet, High-Fat , Insulin Resistance/genetics , Presenilin-1/genetics , Alzheimer Disease/pathology , Amyloid beta-Peptides/genetics , Amyloid beta-Peptides/metabolism , Amyloid beta-Protein Precursor/genetics , Amyloid beta-Protein Precursor/metabolism , Animals , Blotting, Western , Brain/metabolism , Cerebral Cortex/metabolism , Female , Glucose Tolerance Test , Glycogen Synthase Kinase 3/genetics , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Hybridization, Genetic , Hyperglycemia/genetics , Hyperglycemia/pathology , Insulin/blood , Insulin-Like Growth Factor II/genetics , Insulin-Like Growth Factor II/metabolism , Male , Mice , Mice, Transgenic , Phenotype , Phosphorylation , Presenilin-1/metabolism , Signal Transduction
...