Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates
Abstract 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 bra...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-06-01
|
| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12974-023-02826-6 |
| _version_ | 1797806609683972096 |
|---|---|
| author | Marina da Silva Oliveira Larissa Marcely Gomes Cassiano Jeanne Pioline Ketyllen Reis Andrade de Carvalho Anna Christina de Matos Salim Pedro Augusto Alves Gabriel da Rocha Fernandes Alexandre de Magalhães Vieira Machado Roney Santos Coimbra |
| author_facet | Marina da Silva Oliveira Larissa Marcely Gomes Cassiano Jeanne Pioline Ketyllen Reis Andrade de Carvalho Anna Christina de Matos Salim Pedro Augusto Alves Gabriel da Rocha Fernandes Alexandre de Magalhães Vieira Machado Roney Santos Coimbra |
| author_sort | Marina da Silva Oliveira |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-03-13T06:09:52Z |
| format | Article |
| id | doaj.art-deb92487729045b0a0c00d83052368a3 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-03-13T06:09:52Z |
| publishDate | 2023-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-deb92487729045b0a0c00d83052368a32023-06-11T11:20:56ZengBMCJournal of Neuroinflammation1742-20942023-06-0120111410.1186/s12974-023-02826-6Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolatesMarina da Silva Oliveira0Larissa Marcely Gomes Cassiano1Jeanne Pioline2Ketyllen Reis Andrade de Carvalho3Anna Christina de Matos Salim4Pedro Augusto Alves5Gabriel da Rocha Fernandes6Alexandre de Magalhães Vieira Machado7Roney Santos Coimbra8Neurogenômica, Imunopatologia, Instituto René Rachou, FiocruzNeurogenômica, Imunopatologia, Instituto René Rachou, FiocruzNeurogenômica, Imunopatologia, Instituto René Rachou, FiocruzImunologia de Doenças Virais, Instituto René Rachou, FiocruzPlataforma de Sequenciamento NGS (Next Generation Sequencing), Instituto René Rachou, FiocruzImunologia de Doenças Virais, Instituto René Rachou, FiocruzPlataforma de Bioinformática, Instituto René Rachou, FiocruzImunologia de Doenças Virais, Instituto René Rachou, FiocruzNeurogenômica, Imunopatologia, Instituto René Rachou, FiocruzAbstract 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.https://doi.org/10.1186/s12974-023-02826-6Zika virusOrganotypic hippocampal cultureNeurodegenerationNeuroinflammationTranscriptomeChromatin remodeling |
| spellingShingle | Marina da Silva Oliveira Larissa Marcely Gomes Cassiano Jeanne Pioline Ketyllen Reis Andrade de Carvalho Anna Christina de Matos Salim Pedro Augusto Alves Gabriel da Rocha Fernandes Alexandre de Magalhães Vieira Machado Roney Santos Coimbra Organotypic hippocampal culture model reveals differential responses to highly similar Zika virus isolates Journal of Neuroinflammation Zika virus Organotypic hippocampal culture Neurodegeneration Neuroinflammation Transcriptome Chromatin remodeling |
| 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 | Zika virus Organotypic hippocampal culture Neurodegeneration Neuroinflammation Transcriptome Chromatin remodeling |
| url | https://doi.org/10.1186/s12974-023-02826-6 |
| work_keys_str_mv | AT marinadasilvaoliveira organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT larissamarcelygomescassiano organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT jeannepioline organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT ketyllenreisandradedecarvalho organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT annachristinadematossalim organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT pedroaugustoalves organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT gabrieldarochafernandes organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT alexandredemagalhaesvieiramachado organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates AT roneysantoscoimbra organotypichippocampalculturemodelrevealsdifferentialresponsestohighlysimilarzikavirusisolates |