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Title: Comparative Analysis of Retinal Organotypic Cultures and In Vivo Axotomized Retinas
Authors: González-Riquelme, María José
Lucas-Ruiz, Fernando
Galindo-Romero, Caridad
Boia, Raquel 
Ambrósio, António Francisco 
Vidal-Sanz, Manuel
Santiago, Ana Raquel 
Agudo-Barriuso, Marta
Keywords: microglia; Müller cells; retinal ganglion cells; astrocytes; axotomy; in vivo; in vitro
Issue Date: 9-Feb-2023
Publisher: MDPI
Project: This study was supported by the Spanish Ministry of Economy and Competitiveness (project: PID2019-106498GB-I00), Instituto de Salud Carlos III, Fondo Europeo de Desarrollo Regional “Una manera de hacer Europa” (project: PI19/00071), Fundación Séneca, Agencia de Ciencia y Tecnología Región de Murcia (project: 19881/GERM/15); FCT-Portugal (UIDB/04539/2020) and Centro2020 Regional Operational Programme (CENTRO-01-0145-FEDER-000008: BrainHealth 2020). 
Serial title, monograph or event: International Journal of Molecular Sciences
Volume: 24
Issue: 4
Abstract: Retinal organotypic cultures (ROCs) are used as an in vivo surrogate to study retinal ganglion cell (RGC) loss and neuroprotection. In vivo, the gold standard to study RGC degeneration and neuroprotection is optic nerve lesion. We propose here to compare the course of RGC death and glial activation between both models. The left optic nerve of C57BL/6 male mice was crushed, and retinas analyzed from 1 to 9 days after the injury. ROCs were analyzed at the same time points. As a control, intact retinas were used. Retinas were studied anatomically to assess RGC survival, microglial, and macroglial activation. Macroglial and microglial cells showed different morphological activation between models and were activated earlier in ROCs. Furthermore, microglial cell density in the ganglion cell layer was always lower in ROCs than in vivo. RGC loss after axotomy and in vitro followed the same trend up to 5 days. Thereafter, there was an abrupt decrease in viable RGCs in ROCs. However, RGC somas were still immuno-identified by several molecular markers. ROCs are useful for proof-of-concept studies on neuroprotection, but long-term experiments should be carried out in vivo. Importantly, the differential glial activation observed between models and the concomitant death of photoreceptors that occurs in vitro may alter the efficacy of RGC neuroprotective therapies when tested in in vivo models of optic nerve injury.
ISSN: 1422-0067
DOI: 10.3390/ijms24043481
Rights: openAccess
Appears in Collections:I&D CIBB - Artigos em Revistas Internacionais
I&D ICBR - Artigos em Revistas Internacionais
FMUC Medicina - Artigos em Revistas Internacionais

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