Annexin A1 modulation mechanism on the expression of the translocator protein (TSPO)

Abstract

The translocator protein (18kDa) (TSPO) is a binding site for benzodiazepines (BZ) in the Central Nervous System (CNS) and peripheral tissues. Its participation in cancer and inflammatory diseases has been recently evidenced. Annexin A1 (ANXA1) is a protein produced by different cell types and mediates the anti-inflammatory effects of glucocorticoids, especially in neutrophil traffic in the microcirculation and phagocytosis of apoptotic neutrophils by macrophages. We recent showed that TSPO ligands affect the interactions between leukocyte and endothelium and neutrophil chemotaxis induced by G protein coupled receptors (GPCR) and that this can be modulated by endogenous glucocorticoids (GE). This project aims to determine the role of ANXA1 on the function of TSPO in immune cells of the CNS. For this purpose, male, adult, Balb / C mice, wild types or genetically depleted of protein ANXA1 (ANXA1- / -) will be employed. Glial cells (ameboid microglia) and BV2 cells lineage, silenced or with over-expression for ANXA1 will be used to: 1) analysis of the expression of TSPO in basal conditions or in vitro stimulation by lipopolysaccharide (LPS); 2) immunocytochemistry for the location of TSPO in the conditions of the above treatment; 3) RT-PCR (real-time PCR) and Western blot to assess gene and protein expression of TSPO and other mediators of inflammation; 4) evaluation of cytokine expression by flow cytometry flow (CBA - cytometric bead array). This work will contribute to further information undertaken in the laboratory, which aims to elucidate the molecular basis of the control and resolution of the inflammatory process by ANXA1, and to enhance collaboration with Dr. Mauro Perretti and Dr. Egle Solito, both from the William Harvey Research Institute of London. The methodology employed in this project are being has been introduced in the laboratory with the recent visit of Dra. Egle Solito, supported by FAPESP. (AU)