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This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This is the first of two volumes on endosome signaling and includes chapters on such topics as measurement of entry into the endosomal compartment by multi-parametric image analysis, assessment of peptide internalization and endosomal signaling, and VEGF-A in endosomal signaling. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers endosome signalingContain

Endosomes. --- Receptosomes --- Cell organelles --- Cell interaction. --- Enzymology.

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Iron --- Transferrin --- Endocytosis --- Endosomes --- Reticulocytes --- metabolism --- drug effects

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The release of cytokines, chemokines, and other immune-modulating mediators released from innate immune cells, including eosinophils, neutrophils, macrophages, dendritic cells, mast cells, and epithelial cells, is an important event in immunity. Cytokine synthesis and transportation occurs through the canonical protein trafficking pathway associated with endoplasmic reticulum and Golgi. How cytokines are released upon their exit from the trans-Golgi network varies enormously between cell types, and in many cells this has not yet been characterized. This issue delves into the plethora of cytokines released by innate immune cells, and where possible, shines light on specific mechanisms that regulate trafficking and release of Golgi-derived vesicles. Each cell type also shows varying degrees of dependency on microtubule organization and actin cytoskeleton remodeling for cytokine secretion. Understanding the mechanisms of cytokine secretion will reveal the inner workings of individual innate immune cell types, and allow identification of critical regulatory steps in cytokine release.

Cytokines. --- Chemokines. --- secretory granules --- Dendritic Cells --- GTPases --- SNAREs --- Neutrophils --- Epithelial Cells --- degranulation --- Macrophages --- Recycling endosomes --- Eosinophils

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Au cours de ce travail, nous nous sommes intéressés au rôle de vésicules dérivées des mitochondries dans la maturation des endosomes.
Au sein du laboratoire, il a été observé que des protéines de la membrane externe mitochondriale (MEM) étaient recrutées au niveau des endosomes précoces dans des cellules endothéliales en réponse à un traitement au VEGF, un facteur de croissance impliqué de manière majoritaire dans la régulation de l’angiogenèse. Les contacts entre la mitochondrie et des organelles telles que les endosomes, les lysosomes ou la membrane plasmique étant reconnus, nous nous sommes intéressés aux contacts établis entre les endosomes et les mitochondries. Nous avons observé que des contacts pouvaient s’établir via la production de structures semblant dériver des mitochondries (porteuses du marqueur TOM20 typique de la MEM) suite au traitement au VEGF.
La caractérisation du contenu protéique de ces structures suite au développement d’un protocole de purification nous a permis d’identifier différents cargos, dont certains se retrouvaient également au niveau des endosomes précoces après un traitement au VEGF. Nous avons essayé de comprendre le rôle de ces vésicules TOM20+ en ciblant des acteurs nécessaires pour leur biogenèse ou leur trafic intracellulaire (les protéines VPS34 ou Syntaxine 17). Leur diminution d’expression nous a permis d’observer une altération du recrutement endosomal des protéines de la MEM dans ces conditions. Par ailleurs, nous avons pu déterminer que ces protéines VPS34 et Syntaxine 17 sont des inhibiteurs de l’angiogenèse in vitro.
Finalement, nous avons démontré que ces vésicules TOM20+ permettent le recrutement de la protéine RAB5 dans le compartiment endosomal, ce qui permet la maturation des endosomes précoces en endosomes tardifs.

mitochondrie, dynamique mitochondriale, endosomes --- Sciences du vivant > Biochimie, biophysique & biologie moléculaire

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This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This is the second of two volumes on endosome signaling and includes chapters on such topics as measurement of entry into the endosomal compartment by multi-parametric image analysis, assessment of peptide internalization and endosomal signaling, and VEGF-A in endosomal signaling. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers endosome signalingContai

Endosomes. --- Cell interaction. --- Cell-cell interaction --- Cell communication --- Cellular communication (Biology) --- Cellular interaction --- Intercellular communication --- Receptosomes --- Cellular control mechanisms --- Cell organelles --- Enzymology.