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Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates

INTRODUCTION: Zika virus (ZIKV) caused an outbreak in Brazil, in 2015, being associated to microcephaly. ZIKV has a strong neurotropism leading to death of infected cells in different brain regions, including the hippocampus, a major site for neurogenesis. The neuronal populations of the brain are a...

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Autores principales: Oliveira, Marina da Silva, Cassiano, Larissa Marcely Gomes, Pioline, Jeanne, de Carvalho, Ketyllen Reis Andrade, Salim, Anna Christina de Matos, Alves, Pedro Augusto, Fernandes, Gabriel da Rocha, Machado, Alexandre de Magalhães Vieira, Coimbra, Roney Santos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10257278/
https://www.ncbi.nlm.nih.gov/pubmed/37301965
http://dx.doi.org/10.1186/s12974-023-02826-6
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author Oliveira, Marina da Silva
Cassiano, Larissa Marcely Gomes
Pioline, Jeanne
de Carvalho, Ketyllen Reis Andrade
Salim, Anna Christina de Matos
Alves, Pedro Augusto
Fernandes, Gabriel da Rocha
Machado, Alexandre de Magalhães Vieira
Coimbra, Roney Santos
author_facet Oliveira, Marina da Silva
Cassiano, Larissa Marcely Gomes
Pioline, Jeanne
de Carvalho, Ketyllen Reis Andrade
Salim, Anna Christina de Matos
Alves, Pedro Augusto
Fernandes, Gabriel da Rocha
Machado, Alexandre de Magalhães Vieira
Coimbra, Roney Santos
author_sort Oliveira, Marina da Silva
collection PubMed
description INTRODUCTION: Zika virus (ZIKV) caused an outbreak in Brazil, in 2015, being associated to microcephaly. ZIKV has a strong neurotropism leading to death of infected cells in different brain regions, including the hippocampus, a major site for neurogenesis. The neuronal populations of the brain are affected differently by ZIKV from Asian and African ancestral lineages. However, it remains to be investigated whether subtle variations in the ZIKV genome can impact hippocampus infection dynamics and host response. OBJECTIVE: This study evaluated how two Brazilian ZIKV isolates, PE243 and SPH2015, that differ in two specific missense amino acid substitutions, one in the NS1 protein and the other in the NS4A protein, affect the hippocampal phenotype and transcriptome. METHODS: Organotypic hippocampal cultures (OHC) from infant Wistar rats were infected with PE243 or SPH2015 and analyzed in time series using immunofluorescence, confocal microscopy, RNA-Seq and RT-qPCR. RESULTS: Unique patterns of infection and changes in neuronal density in the OHC were observed for PE243 and SPH2015 between 8 and 48 h post infection (p.i.). Phenotypic analysis of microglia indicated that SPH2015 has a greater capacity for immune evasion. Transcriptome analysis of OHC at 16 h p.i. disclosed 32 and 113 differentially expressed genes (DEGs) in response to infection with PE243 and SPH2015, respectively. Functional enrichment analysis suggested that infection with SPH2015 activates mostly astrocytes rather than microglia. PE243 downregulated biological process of proliferation of brain cells and upregulated those associated with neuron death, while SPH2015 downregulated processes related to neuronal development. Both isolates downregulated cognitive and behavioral development processes. Ten genes were similarly regulated by both isolates. They are putative biomarkers of early hippocampus response to ZIKV infection. At 5, 7, and 10 days p.i., neuronal density of infected OHC remained below controls, and mature neurons of infected OHC showed an increase in the epigenetic mark H3K4me3, which is associated to a transcriptionally active state. This feature is more prominent in response to SPH2015. CONCLUSION: Subtle genetic diversity of the ZIKV affects the dynamics of viral dissemination in the hippocampus and host response in the early stages of infection, which may lead to different long-term effects in neuronal population. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12974-023-02826-6.
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spelling pubmed-102572782023-06-11 Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates Oliveira, Marina da Silva Cassiano, Larissa Marcely Gomes Pioline, Jeanne de Carvalho, Ketyllen Reis Andrade Salim, Anna Christina de Matos Alves, Pedro Augusto Fernandes, Gabriel da Rocha Machado, Alexandre de Magalhães Vieira Coimbra, Roney Santos J Neuroinflammation Research INTRODUCTION: Zika virus (ZIKV) caused an outbreak in Brazil, in 2015, being associated to microcephaly. ZIKV has a strong neurotropism leading to death of infected cells in different brain regions, including the hippocampus, a major site for neurogenesis. The neuronal populations of the brain are affected differently by ZIKV from Asian and African ancestral lineages. However, it remains to be investigated whether subtle variations in the ZIKV genome can impact hippocampus infection dynamics and host response. OBJECTIVE: This study evaluated how two Brazilian ZIKV isolates, PE243 and SPH2015, that differ in two specific missense amino acid substitutions, one in the NS1 protein and the other in the NS4A protein, affect the hippocampal phenotype and transcriptome. METHODS: Organotypic hippocampal cultures (OHC) from infant Wistar rats were infected with PE243 or SPH2015 and analyzed in time series using immunofluorescence, confocal microscopy, RNA-Seq and RT-qPCR. RESULTS: Unique patterns of infection and changes in neuronal density in the OHC were observed for PE243 and SPH2015 between 8 and 48 h post infection (p.i.). Phenotypic analysis of microglia indicated that SPH2015 has a greater capacity for immune evasion. Transcriptome analysis of OHC at 16 h p.i. disclosed 32 and 113 differentially expressed genes (DEGs) in response to infection with PE243 and SPH2015, respectively. Functional enrichment analysis suggested that infection with SPH2015 activates mostly astrocytes rather than microglia. PE243 downregulated biological process of proliferation of brain cells and upregulated those associated with neuron death, while SPH2015 downregulated processes related to neuronal development. Both isolates downregulated cognitive and behavioral development processes. Ten genes were similarly regulated by both isolates. They are putative biomarkers of early hippocampus response to ZIKV infection. At 5, 7, and 10 days p.i., neuronal density of infected OHC remained below controls, and mature neurons of infected OHC showed an increase in the epigenetic mark H3K4me3, which is associated to a transcriptionally active state. This feature is more prominent in response to SPH2015. CONCLUSION: Subtle genetic diversity of the ZIKV affects the dynamics of viral dissemination in the hippocampus and host response in the early stages of infection, which may lead to different long-term effects in neuronal population. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12974-023-02826-6. BioMed Central 2023-06-10 /pmc/articles/PMC10257278/ /pubmed/37301965 http://dx.doi.org/10.1186/s12974-023-02826-6 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Oliveira, Marina da Silva
Cassiano, Larissa Marcely Gomes
Pioline, Jeanne
de Carvalho, Ketyllen Reis Andrade
Salim, Anna Christina de Matos
Alves, Pedro Augusto
Fernandes, Gabriel da Rocha
Machado, Alexandre de Magalhães Vieira
Coimbra, Roney Santos
Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
title Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
title_full Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
title_fullStr Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
title_full_unstemmed Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
title_short Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
title_sort organotypic hippocampal culture model reveals differential responses to highly similar zika virus isolates
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10257278/
https://www.ncbi.nlm.nih.gov/pubmed/37301965
http://dx.doi.org/10.1186/s12974-023-02826-6
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