Methylphenidate-triggered ROS generation promotes caveolae-mediated transcytosis via Rac1 signaling and c-Src-dependent caveolin-1 phosphorylation in human brain endothelial cells

• Type: Article in International Scientific Journal
• Date: 2016

Authors

Coelho-Santos, V (Author) Other; Socodato, R (Author) Other; Portugal, C (Author) Other; Leitão, RA (Author) Other; Rito, M (Author) Other; Barbosa, M (Author) Other; Couraud, P (Author) Other; Romero, IA (Author) Other; Weksler, B (Author) Other; Minshall, RD (Author) Other; Fontes-Ribeiro, C (Author) Other; Summavielle, T (Author) Other; Relvas, JB (Author) ICBAS; Silva, AP (Author) Other

Journal

Title: Cellular and Molecular Life Sciences

Pages: 1-16

ISSN: 1420-682X

Publisher: Springer Nature

Indexing

Scopus - 0 Citations

Other information

Authenticus ID: P-00K-YBS

DOI: 10.1007/s00018-016-2301-3

Abstract (EN): Methylphenidate (MPH) is an amphetamine-like stimulant commonly prescribed for attention deficit hyperactivity disorder. Despite its widespread use, the cellular/molecular effects of MPH remain elusive. Here, we report a novel direct role of MPH on the regulation of macromolecular flux through human brain endothelial cells (ECs). MPH significantly increased caveolae-mediated transcytosis of horseradish peroxidase through ECs without affecting paracellular permeability. Using FRET-based live cell imaging, together with pharmacological inhibitors and lentiviral-mediated shRNA knockdown, we demonstrate that MPH promoted ROS generation via activation of Rac1-dependent NADPH oxidase (NOX) and c-Src activation at the plasma membrane. c-Src in turn was shown to mediate the phosphorylation of caveolin-1 (Cav1) on Tyr14 leading to enhanced caveolae formation and transendothelial transport. Accordingly, the inhibition of Cav1 phosphorylation by overexpression of a phosphodefective Cav1Y14F mutant or knocking down Cav1 expression abrogated MPH-induced transcytosis. In addition, both vitamin C and inhibition of NOX blocked MPH-triggered vesicular transport. This study, therefore, identifies Rac1/NOX/c-Src-dependent signaling in MPH-induced increase in transendothelial permeability of brain endothelial cell monolayers via caveolae-mediated transcytosis. © 2016 Springer International Publishing

Language: English

Type (Professor's evaluation): Scientific