Activation of microglial cells triggers a release of brain-derived neurotrophic factor (BDNF) inducing their proliferation in an adenosine A2A receptor-dependent manner: A2A receptor blockade prevents BDNF release and proliferation of microglia

Background: Brain-derived neurotrophic factor (BDNF) has been shown to control microglial responses in neuropathic pain. Since adenosine A_2A receptors (A_2ARs) control neuroinflammation, as well as the production and function of BDNF, we tested to see if A_2AR controls the microglia-dependent secretion of BDNF and the proliferation of microglial cells, a crucial event in neuroinflammation. Methods: Murine N9 microglial cells were challenged with lipopolysaccharide (LPS, 100 ng/mL) in the absence or in the presence of the A_2AR antagonist, SCH58261 (50 nM), as well as other modulators of A_2AR signaling. The BDNF cellular content and secretion were quantified by Western blotting and ELISA, A_2AR density was probed by Western blotting and immunocytochemistry and cell proliferation was assessed by BrdU incorporation. Additionally, the A_2AR modulation of LPS-driven cell proliferation was also tested in primary cultures of mouse microglia. Results: LPS induced time-dependent changes of the intra- and extracellular levels of BDNF and increased microglial proliferation. The maximal LPS-induced BDNF release was time-coincident with an LPS-induced increase of the A_2AR density. Notably, removing endogenous extracellular adenosine or blocking A_2AR prevented the LPS-mediated increase of both BDNF secretion and proliferation, as well as exogenous BDNF-induced proliferation. Conclusions: We conclude that A_2AR activation plays a mandatory role controlling the release of BDNF from activated microglia, as well as the autocrine/paracrine proliferative role of BDNF. [ABSTRACT FROM AUTHOR]