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Subcellular localization of Oropouche virus in human brain slice cultures using super-resolution microscopy

Grant number: 23/01202-8
Support Opportunities:Scholarships abroad - Research Internship - Doctorate
Effective date (Start): July 01, 2023
Effective date (End): December 31, 2023
Field of knowledge:Biological Sciences - Biochemistry - Chemistry of Macromolecules
Principal Investigator:Adriano Silva Sebollela
Grantee:Glaucia Maria de Almeida
Supervisor: Akira Ono
Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Research place: University of Michigan, United States  
Associated to the scholarship:21/12263-2 - Tissue-molecular alterations associated with viral infections in adult human brain slice cultures, BP.DR

Abstract

Brain slice cultures maintain the cytoarchitecture and cell connections present in brain tissue and have been widely applied in neuroscience research. In our laboratory, we have developed a method for culturing adult human brain tissue under free-floating conditions. In the past few years, we have applied this model to study cellular and molecular aspects associated with the brain pathogenicity of emerging neurotropic viruses like the Oropouche virus (OROV), an arbovirus that causes a febrile illness endemic in South America. Recent studies have reported the presence of OROV in cerebrospinal fluid in patients with meningoencephalitis, as well as neurological symptoms such as diplopia and nystagmus. Using the human brain slice model, we showed for the first time that OROV infects and replicates in neural cells obtained from adult donors. We observed that OROV preferentially infects microglia but also infects neurons to a lesser extent. Interestingly, the distribution of OROV proteins seems to be different in microglia and neurons, suggesting that OROV assembly can be mechanistically distinct in each cell type. Motivated by these findings, we now intend to focus on determining the subcellular localization of OROV particles/proteins in infected cells in brain slices using super-resolution imaging. The super-resolution will be obtained using expansion microscopy, a cutting-edge technique that allows super-resolution imaging in thick samples like brain slices. We anticipate this approach will significantly contribute to advancing my PhD project, and furthermore the field's knowledge of the biology of Oropouche virus infection in the human CNS. (AU)

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