ANALYSIS OF MITOCHONDRIAL COMPLEX I SUBUNIT (NDUFS2) ROLE IN PRIMARY CORTICAL NEURONS FROM A SCHIZOPHRENIA-LIKE ANIMAL MODEL

• Grant number: 23/14145-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: June 01, 2024
• End date: May 31, 2025
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: Cristoforo Scavone
• Grantee: Amanda Siena dos Santos
• Supervisor: Juan-Carlos Leza
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: Universidad Complutense de Madrid (UCM), Spain
• Associated to the scholarship: 21/03021-5 - The role of the mitochondrial function on synaptic dynamics and neurogenesis in cells exposed to rotenone: relevance for psychiatric disorders, BP.DR

Abstract

Schizophrenia (SCZ) is a disabling psychiatric disorder that affects around 1% of the global population. Patients with SCZ present positive, negative, and cognitive symptoms. The SCZ onset occurs at the beginning of adulthood, with correlates with the late stages of neurodevelopment. Then, nowadays, SCZ has been studied as a neurodevelopmental disorder. Some disturbances during neurodevelopment can increase the risk of developing SCZ, such as maternal viral and bacterial infections, and perinatal complications. As a multifaceted disorder, the development of in vitro and in vivo models seems indispensable in order to better understand SCZ pathophysiology. Animal models have been developed to study the SCZ onset and disorder evolution. For example, the mimic of viral infection during the last trimester of pregnancy (embryonic day 15), with PolyI:C administration in Wistar rats, produces offspring with SCZ-like phenotype. Indeed, previous results of our research group showed that mitochondrial complex I inhibition, with Rotenone (Rot) administration, in Wistar rats during neurodevelopment (from 5 to 11 days old) can lead to an SCZ-like phenotype. The Rot treatment in primary cortical neurons with the same amount present in the Rot-treated animal model (1.325nM) leads to neuronal impairment in mitochondria respiration, superoxide production, and dendritic arborization. However, more studies are necessary to fully elucidate the Rot modifications in neurons and their relationship with SCZ cellular phenotype. Therefore, this project aims to compare the neuronal abnormalities in the primary cortical neurons from an SCZ animal model (PolyI:C administration), a kind of culture that does not have reports in the literature, with the primary cortical neurons treated with Rot. Nevertheless, investigate if the NDUFS2 CI subunit is involved in the cellular disruptions promoted by Rot treatment on the cortical neurons, and if the PolyI:C neuronal phenotype can be worsened or modified by Rot treatment and/or NDUFS2 silencing. For this, we will use embryos 19 days old from PolyI:C-treated Wistar rats and treated-control Wistar rats to obtain a primary cortical neuronal culture. After that, we will treat both cells with siRNA for Ndufs2 and Rot 1.325nM. Will be evaluated: i) mitochondrial superoxide production, by MitoSox probe; ii) mitochondrial supercomplex assay, by native gel electrophoresis; iii) mitochondrial dynamics, by immunocytochemistry; iv) antioxidant cascade activation, by western blot; v) dendritic arborization, by immunocytochemistry; vi) synaptic structuration, by immunocytochemistry. With this project, we expect to increase the understanding of the mitochondrial complex I role during neuronal development and the cellular phenotype of an SCZ-like animal model.