Evaluation of neurite desnsity in neurons with mutations in the LRRK2 and PRKN genes

• Grant number: 25/02168-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2025
• End date: May 31, 2026
• Field of knowledge: Health Sciences - Medicine - Psychiatry
• Principal Investigator: Patrícia Maria de Carvalho Aguiar
• Grantee: Victoria Alves Fulaneto
• Host Institution: Instituto Israelita de Ensino e Pesquisa Albert Einstein (IIEPAE). Sociedade Beneficente Israelita Brasileira Albert Einstein (SBIBAE). São Paulo , SP, Brazil

Abstract

Introduction: Parkinson's Disease (PD) is a multifactorial neurodegenerative condition with no known cure. Mutations in the LRRK2 and PARK2 genes affect cellular processes and accelerate neurodegeneration, in addition to altering the morphology and density of neurites in dopaminergic neurons. Studies suggest that degeneration of dopaminergic terminals precedes the death of neurons in the substantia nigra. In this study, we will compare the density and morphology of neurites between dopaminergic neurons with mutations in the LRRK2 and PARK2 genes and healthy controls.Methods: Neuroprogenitor cell (NPC) lines developed in our laboratory from human skin fibroblast-derived induced pluripotent stem cells will be differentiated into dopaminergic neurons. In the first stage of this project, we will test NPC differentiation protocols for dopaminergic neurons in healthy control cells. Once the most effective method is established, healthy lines and lines with mutations in the LRRK2 and PARK2 genes will be differentiated into dopaminergic neurons and maintained in culture for 4 to 8 weeks. Subsequently, immunohistochemistry and western blot will be performed for neural antibodies, including tyrosine hydroxylase, and confocal microscopy images will be analyzed using the neurolucida software to quantify neurite density.