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  1. Article ; Online: Protocol for observing microtubules and microtubule ends in both fixed and live primary microglia cells.

    Sanchini, Caterina / Rosito, Maria / Comincini, Alessandro / De Panfilis, Simone / Bartolini, Francesca / Di Angelantonio, Silvia

    STAR protocols

    2023  Volume 4, Issue 3, Page(s) 102499

    Abstract: Microtubule dynamics and orientation have crucial roles in many vital cellular processes. However, functional live imaging of microtubules and/or microtubule ends in primary microglia can be challenging. Here, we present a protocol for observing ... ...

    Abstract Microtubule dynamics and orientation have crucial roles in many vital cellular processes. However, functional live imaging of microtubules and/or microtubule ends in primary microglia can be challenging. Here, we present a protocol for observing microtubules and microtubule ends in both fixed and live primary microglia cells. We describe steps for microglia culture and in vitro stimulation, SiR-tubulin labeling, lentivirus preparation, live imaging, immunostaining, and image acquisition. We also provide procedures for SiR-tubulin, EB3-EGFP, and EB1 analyses. For complete details on the use and execution of this protocol, please refer to Rosito et al. (2023).
    MeSH term(s) Tubulin/genetics ; Microtubule-Associated Proteins/genetics ; Microglia ; Microtubules
    Chemical Substances Tubulin ; Microtubule-Associated Proteins
    Language English
    Publishing date 2023-08-12
    Publishing country United States
    Document type Journal Article
    ISSN 2666-1667
    ISSN (online) 2666-1667
    DOI 10.1016/j.xpro.2023.102499
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Unlocking Neural Function with 3D In Vitro Models: A Technical Review of Self-Assembled, Guided, and Bioprinted Brain Organoids and Their Applications in the Study of Neurodevelopmental and Neurodegenerative Disorders.

    D'Antoni, Chiara / Mautone, Lorenza / Sanchini, Caterina / Tondo, Lucrezia / Grassmann, Greta / Cidonio, Gianluca / Bezzi, Paola / Cordella, Federica / Di Angelantonio, Silvia

    International journal of molecular sciences

    2023  Volume 24, Issue 13

    Abstract: Understanding the complexities of the human brain and its associated disorders poses a significant challenge in neuroscience. Traditional research methods have limitations in replicating its intricacies, necessitating the development of in vitro models ... ...

    Abstract Understanding the complexities of the human brain and its associated disorders poses a significant challenge in neuroscience. Traditional research methods have limitations in replicating its intricacies, necessitating the development of in vitro models that can simulate its structure and function. Three-dimensional in vitro models, including organoids, cerebral organoids, bioprinted brain models, and functionalized brain organoids, offer promising platforms for studying human brain development, physiology, and disease. These models accurately replicate key aspects of human brain anatomy, gene expression, and cellular behavior, enabling drug discovery and toxicology studies while providing insights into human-specific phenomena not easily studied in animal models. The use of human-induced pluripotent stem cells has revolutionized the generation of 3D brain structures, with various techniques developed to generate specific brain regions. These advancements facilitate the study of brain structure development and function, overcoming previous limitations due to the scarcity of human brain samples. This technical review provides an overview of current 3D in vitro models of the human cortex, their development, characterization, and limitations, and explores the state of the art and future directions in the field, with a specific focus on their applications in studying neurodevelopmental and neurodegenerative disorders.
    MeSH term(s) Animals ; Humans ; Brain/metabolism ; Neurodegenerative Diseases/metabolism ; Organoids ; Induced Pluripotent Stem Cells
    Language English
    Publishing date 2023-06-28
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms241310762
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  3. Article ; Online: Microglia reactivity entails microtubule remodeling from acentrosomal to centrosomal arrays.

    Rosito, Maria / Sanchini, Caterina / Gosti, Giorgio / Moreno, Manuela / De Panfilis, Simone / Giubettini, Maria / Debellis, Doriana / Catalano, Federico / Peruzzi, Giovanna / Marotta, Roberto / Indrieri, Alessia / De Leonibus, Elvira / De Stefano, Maria Egle / Ragozzino, Davide / Ruocco, Giancarlo / Di Angelantonio, Silvia / Bartolini, Francesca

    Cell reports

    2023  Volume 42, Issue 2, Page(s) 112104

    Abstract: Microglia reactivity entails a large-scale remodeling of cellular geometry, but the behavior of the microtubule cytoskeleton during these changes remains unexplored. Here we show that activated microglia provide an example of microtubule reorganization ... ...

    Abstract Microglia reactivity entails a large-scale remodeling of cellular geometry, but the behavior of the microtubule cytoskeleton during these changes remains unexplored. Here we show that activated microglia provide an example of microtubule reorganization from a non-centrosomal array of parallel and stable microtubules to a radial array of more dynamic microtubules. While in the homeostatic state, microglia nucleate microtubules at Golgi outposts, and activating signaling induces recruitment of nucleating material nearby the centrosome, a process inhibited by microtubule stabilization. Our results demonstrate that a hallmark of microglia reactivity is a striking remodeling of the microtubule cytoskeleton and suggest that while pericentrosomal microtubule nucleation may serve as a distinct marker of microglia activation, inhibition of microtubule dynamics may provide a different strategy to reduce microglia reactivity in inflammatory disease.
    MeSH term(s) Microglia ; Microtubules ; Centrosome ; Cytoskeleton ; Golgi Apparatus ; Tubulin
    Chemical Substances Tubulin
    Language English
    Publishing date 2023-02-13
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2023.112104
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  4. Article: Substrate stiffness effect on molecular crosstalk of epithelial-mesenchymal transition mediators of human glioblastoma cells.

    Basilico, Bernadette / Palamà, Ilaria Elena / D'Amone, Stefania / Lauro, Clotilde / Rosito, Maria / Grieco, Maddalena / Ratano, Patrizia / Cordella, Federica / Sanchini, Caterina / Di Angelantonio, Silvia / Ragozzino, Davide / Cascione, Mariafrancesca / Gigli, Giuseppe / Cortese, Barbara

    Frontiers in oncology

    2022  Volume 12, Page(s) 983507

    Abstract: The complexity of the microenvironment effects on cell response, show accumulating evidence that glioblastoma (GBM) migration and invasiveness are influenced by the mechanical rigidity of their surroundings. The epithelial-mesenchymal transition (EMT) is ...

    Abstract The complexity of the microenvironment effects on cell response, show accumulating evidence that glioblastoma (GBM) migration and invasiveness are influenced by the mechanical rigidity of their surroundings. The epithelial-mesenchymal transition (EMT) is a well-recognized driving force of the invasive behavior of cancer. However, the primary mechanisms of EMT initiation and progression remain unclear. We have previously showed that certain substrate stiffness can selectively stimulate human GBM U251-MG and GL15 glioblastoma cell lines motility. The present study unifies several known EMT mediators to uncover the reason of the regulation and response to these stiffnesses. Our results revealed that changing the rigidity of the mechanical environment tuned the response of both cell lines through change in morphological features, epithelial-mesenchymal markers (E-, N-Cadherin), EGFR and ROS expressions in an interrelated manner. Specifically, a stiffer microenvironment induced a mesenchymal cell shape, a more fragmented morphology, higher intracellular cytosolic ROS expression and lower mitochondrial ROS. Finally, we observed that cells more motile showed a more depolarized mitochondrial membrane potential. Unravelling the process that regulates GBM cells' infiltrative behavior could provide new opportunities for identification of new targets and less invasive approaches for treatment.
    Language English
    Publishing date 2022-08-25
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2649216-7
    ISSN 2234-943X
    ISSN 2234-943X
    DOI 10.3389/fonc.2022.983507
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  5. Article ; Online: Antibiotics Treatment Modulates Microglia-Synapses Interaction.

    Cordella, Federica / Sanchini, Caterina / Rosito, Maria / Ferrucci, Laura / Pediconi, Natalia / Cortese, Barbara / Guerrieri, Francesca / Pascucci, Giuseppe Rubens / Antonangeli, Fabrizio / Peruzzi, Giovanna / Giubettini, Maria / Basilico, Bernadette / Pagani, Francesca / Grimaldi, Alfonso / D'Alessandro, Giuseppina / Limatola, Cristina / Ragozzino, Davide / Di Angelantonio, Silvia

    Cells

    2021  Volume 10, Issue 10

    Abstract: Dysbiosis' of the adult gut microbiota, in response to challenges such as infection, altered diet, stress, and antibiotics treatment has been recently linked to pathological alteration of brain function and behavior. Moreover, gut microbiota composition ...

    Abstract 'Dysbiosis' of the adult gut microbiota, in response to challenges such as infection, altered diet, stress, and antibiotics treatment has been recently linked to pathological alteration of brain function and behavior. Moreover, gut microbiota composition constantly controls microglia maturation, as revealed by morphological observations and gene expression analysis. However, it is unclear whether microglia functional properties and crosstalk with neurons, known to shape and modulate synaptic development and function, are influenced by the gut microbiota. Here, we investigated how antibiotic-mediated alteration of the gut microbiota influences microglial and neuronal functions in adult mice hippocampus. Hippocampal microglia from adult mice treated with oral antibiotics exhibited increased microglia density, altered basal patrolling activity, and impaired process rearrangement in response to damage. Patch clamp recordings at CA3-CA1 synapses revealed that antibiotics treatment alters neuronal functions, reducing spontaneous postsynaptic glutamatergic currents and decreasing synaptic connectivity, without reducing dendritic spines density. Antibiotics treatment was unable to modulate synaptic function in CX3CR1-deficient mice, pointing to an involvement of microglia-neuron crosstalk through the CX3CL1/CX3CR1 axis in the effect of dysbiosis on neuronal functions. Together, our findings show that antibiotic alteration of gut microbiota impairs synaptic efficacy, suggesting that CX3CL1/CX3CR1 signaling supporting microglia is a major player in in the gut-brain axis, and in particular in the gut microbiota-to-neuron communication pathway.
    MeSH term(s) Animals ; Anti-Bacterial Agents/pharmacology ; CX3C Chemokine Receptor 1/metabolism ; Chemokine CX3CL1/metabolism ; Gene Expression Regulation/drug effects ; Glutamic Acid/metabolism ; Hippocampus/pathology ; Inflammation/genetics ; Mice ; Microglia/drug effects ; Microglia/metabolism ; Neurons/drug effects ; Neurons/metabolism ; Signal Transduction/drug effects ; Synapses/drug effects ; Synapses/metabolism ; Synaptic Transmission/drug effects
    Chemical Substances Anti-Bacterial Agents ; CX3C Chemokine Receptor 1 ; Chemokine CX3CL1 ; Glutamic Acid (3KX376GY7L)
    Language English
    Publishing date 2021-10-04
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells10102648
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  6. Article ; Online: Microglia control glutamatergic synapses in the adult mouse hippocampus.

    Basilico, Bernadette / Ferrucci, Laura / Ratano, Patrizia / Golia, Maria T / Grimaldi, Alfonso / Rosito, Maria / Ferretti, Valentina / Reverte, Ingrid / Sanchini, Caterina / Marrone, Maria C / Giubettini, Maria / De Turris, Valeria / Salerno, Debora / Garofalo, Stefano / St-Pierre, Marie-Kim / Carrier, Micael / Renzi, Massimiliano / Pagani, Francesca / Modi, Brijesh /
    Raspa, Marcello / Scavizzi, Ferdinando / Gross, Cornelius T / Marinelli, Silvia / Tremblay, Marie-Ève / Caprioli, Daniele / Maggi, Laura / Limatola, Cristina / Di Angelantonio, Silvia / Ragozzino, Davide

    Glia

    2021  Volume 70, Issue 1, Page(s) 173–195

    Abstract: Microglia cells are active players in regulating synaptic development and plasticity in the brain. However, how they influence the normal functioning of synapses is largely unknown. In this study, we characterized the effects of pharmacological microglia ...

    Abstract Microglia cells are active players in regulating synaptic development and plasticity in the brain. However, how they influence the normal functioning of synapses is largely unknown. In this study, we characterized the effects of pharmacological microglia depletion, achieved by administration of PLX5622, on hippocampal CA3-CA1 synapses of adult wild type mice. Following microglial depletion, we observed a reduction of spontaneous and evoked glutamatergic activity associated with a decrease of dendritic spine density. We also observed the appearance of immature synaptic features and higher levels of plasticity. Microglia depleted mice showed a deficit in the acquisition of the Novel Object Recognition task. These events were accompanied by hippocampal astrogliosis, although in the absence ofneuroinflammatory condition. PLX-induced synaptic changes were absent in Cx3cr1
    MeSH term(s) Animals ; Brain ; Excitatory Amino Acid Agents/pharmacology ; Hippocampus ; Mice ; Microglia ; Neurons ; Organic Chemicals/pharmacology ; Synapses/physiology
    Chemical Substances Excitatory Amino Acid Agents ; Organic Chemicals ; PLX5622
    Language English
    Publishing date 2021-10-18
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 639414-0
    ISSN 1098-1136 ; 0894-1491
    ISSN (online) 1098-1136
    ISSN 0894-1491
    DOI 10.1002/glia.24101
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