Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants
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AutorFerrer-Martín, Rosa María; Martín-Oliva, David; Sierra, Ana; Carrasco Sierra, María del Carmen; Martín-Estebané, María; Calvente Iglesias, Ruth; Martín-Guerrero, Sandra M.; Marín-Teva, José Luis; Navascués Martínez, Julio; Cuadros Ojeda, Miguel Ángel
Public Library of Science (PLOS)
Microglial cellsRetinaFlow cytometryPhtoreceptorsCell deathMacrophagesRetinal degenerationLiposomes
Ferrer-Martín, R.M.; et al. Microglial Activation Promotes Cell Survival in Organotypic Cultures of Postnatal Mouse Retinal Explants. Plos One, 10(8): e0135238 (2015). [http://hdl.handle.net/10481/37271]
PatrocinadorThis work was supported by Junta de Andalucía, Spain, Grant P07-CVI-03008 (http://www.juntadeandalucia.es/organismos/economiainnovacioncienciayempleo.html), and Ministerio de Economía y Competitividad, Spain, Grant BFU2010-19981 (http://www.mineco.gob.es/portal/site/mineco/idi).
The role of microglia during neurodegeneration remains controversial. We investigated whether microglial cells have a neurotoxic or neuroprotective function in the retina. Retinal explants from 10-day-old mice were treated in vitro with minocycline to inhibit microglial activation, with LPS to increase microglial activation, or with liposomes loaded with clodronate (Lip-Clo) to deplete microglial cells. Flow cytometry was used to assess the viability of retinal cells in the explants and the TUNEL method to show the distribution of dead cells. The immunophenotypic and morphological features of microglia and their distribution were analyzed with flow cytometry and immunocytochemistry. Treatment of retinal explants with minocycline reduced microglial activation and simultaneously significantly decreased cell viability and increased the presence of TUNEL-labeled cell profiles. This treatment also prevented the migration of microglial cells towards the outer nuclear layer, where cell death was most abundant. The LPS treatment increased microglial activation but had no effect on cell viability or microglial distribution. Finally, partial microglial removal with Lip-Clo diminished the cell viability in the retinal explants, showing a similar effect to that of minocycline. Hence, cell viability is diminished in retinal explants cultured in vitro when microglial cells are removed or their activation is inhibited, indicating a neurotrophic role for microglia in this system.