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Open AccessArticle
A New Ex Vivo Model Based on Mouse Retinal Explants for the Study of Ocular Toxoplasmosis
by
Veronica Rodriguez Fernandez
Veronica Rodriguez Fernandez 1,2,†
,
Rosario Amato
Rosario Amato 3,†
,
Simona Piaggi
Simona Piaggi 1,
Barbara Pinto
Barbara Pinto 1,
Giovanni Casini
Giovanni Casini 3,4,*
and
Fabrizio Bruschi
Fabrizio Bruschi 1,*
1
Department of Translational Research, School of Medicine, University of Pisa, 56126 Pisa, Italy
2
Department of Infectious Diseases and Public Health, La Sapienza University, 00185 Rome, Italy
3
Department of Biology, University of Pisa, 56126 Pisa, Italy
4
Interdepartmental Research Center Nutrafood “Nutraceuticals and Food for Health”, University of Pisa, 56126 Pisa, Italy
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Pathogens 2024, 13(8), 701; https://doi.org/10.3390/pathogens13080701 (registering DOI)
Submission received: 27 June 2024
/
Revised: 5 August 2024
/
Accepted: 18 August 2024
/
Published: 19 August 2024
Abstract
Ocular toxoplasmosis is the most prevalent clinical manifestation of T. gondii infection, which causes irreversible retinal damage. Different experimental models have been developed to study this pathology. In the present study, a new, ex vivo model is proposed to contribute to the elucidation of disease mechanisms and to possible therapeutic solutions. Ex-vivo retinal explants, prepared from mouse retinas following established protocols, were incubated with T. gondii tachyzoites maintained in Vero cells. At different times, starting at 12 h up to 10 days of incubation, the explants were analyzed with immunofluorescence and Western blot to investigate their responses to parasite infection. T. gondii invasion of the retinal thickness was evident after 3 days in culture, where parasites could be detected around retinal cell nuclei. This was paralleled by putative cyst formation and microglial activation. At the same time, an evident increase in inflammatory and oxidative stress markers was detected in infected explants compared to controls. Cell death also appeared to occur in retinal explants after 3 days of T. gondii infection, and it was characterized by increased necroptotic but not apoptotic markers. The proposed model recapitulates the main characteristics of T. gondii retinal infection within 3 days of incubation and, therefore, allows for studying the very early events of the process. In addition, it requires only a limited number of animals and offers easy manipulation and accessibility for setting up different experimental conditions and assessing the effects of putative drugs for therapy.
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MDPI and ACS Style
Rodriguez Fernandez, V.; Amato, R.; Piaggi, S.; Pinto, B.; Casini, G.; Bruschi, F.
A New Ex Vivo Model Based on Mouse Retinal Explants for the Study of Ocular Toxoplasmosis. Pathogens 2024, 13, 701.
https://doi.org/10.3390/pathogens13080701
AMA Style
Rodriguez Fernandez V, Amato R, Piaggi S, Pinto B, Casini G, Bruschi F.
A New Ex Vivo Model Based on Mouse Retinal Explants for the Study of Ocular Toxoplasmosis. Pathogens. 2024; 13(8):701.
https://doi.org/10.3390/pathogens13080701
Chicago/Turabian Style
Rodriguez Fernandez, Veronica, Rosario Amato, Simona Piaggi, Barbara Pinto, Giovanni Casini, and Fabrizio Bruschi.
2024. "A New Ex Vivo Model Based on Mouse Retinal Explants for the Study of Ocular Toxoplasmosis" Pathogens 13, no. 8: 701.
https://doi.org/10.3390/pathogens13080701
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