Analysis of the consequences of Argonauta-2 protein knockdown on synaptogenesis in vitro

• Grant number: 21/03711-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2021
• End date: December 31, 2021
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Alexandre Hiroaki Kihara
• Grantee: Gabrieli Bovi dos Santos
• Host Institution: Centro de Matemática, Computação e Cognição (CMCC). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

The retina has a complex morphology and is composed of more than 60 cell types highly specialized for the transduction and modulation of light information, and the transmission of this information is coordinated by chemical and electrical synapses. A key factor in cell development, cell maturation, and synaptogenesis are small non-coding RNAs, also known as microRNAs (miRNAs). The miRNAs are responsible for the regulation of gene expression through interaction with complementary messenger RNAs (mRNA). In their action process, the RNAs-induced silencing complex (RISC) stands out, composed of several proteins, among them the Agonauta-2 protein (AGO2), which guides the miRNA to the target mRNA, and may act in the cleavage or degradation of the target mRNA. Recent studies show other functions attributed to AGO2, such as active role in axonal development and regeneration and in the formation of growth cones. However, the role of AGO2 in retinal synaptogenesis is not yet fully elucidated. Thus, the aim of this project is to analyze the consequences of AGO2 protein knockdown on invitro synaptogenesis. For this, primary retinal cell cultures will be transfected with the AGO2 inhibition plasmid (pLKO.1puro-AGO2), and the consequences on synaptogenesis will be evaluated by immunofluorescence and western blotting. With this project, we intend to evaluate the distribution of synaptic proteins, morphology of dendrites and axons, after knockdown of AGO2 protein in retinal cells in vitro. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MOVIO, MARILIA INES; DE LIMA-VASCONCELLOS, THEO HENRIQUE; DOS SANTOS, GABRIELI BOVI; ECHEVERRY, MARCELA BERMUDEZ; COLOMBO, ELISABETTA; MATTOS, LEONARDO S.; RESENDE, RODRIGO RIBEIRO; KIHARA, ALEXANDRE HIROAKI. Retinal organoids from human-induced pluripotent stem cells: From studying retinal dystrophies to early diagnosis of Alzheimer?s and Parkinson?s disease. SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY, v. 144, p. 10-pg., 2023-03-17. (21/14227-3, 21/03711-1, 19/17892-8, 20/02035-0, 20/11667-0, 21/11969-9, 17/26388-6)