Exploiting Nanotechnologies and TRPV1 Channels to Investigate the Putative Anandamide Membrane Transporter
Figure 4
Characterization of anandamide release from PCL-NPs and evidence that PCL-NP rapidly enter HEK-293 cells over-expressing the human recombinant TRPV1 receptor.
(A) Anandamide release from PCL-NP-AEA at 37°C and at a 99.9% dilution in distilled water (0.5 µl stock suspension in 500 µl water). AEA was measured by HPLC. In a separate experiment, ∼80% of AEA was retained by PCL-NP-AEA incubated for 1 min with the assay buffer used for the experiments of Figs. 1,3. Each bar indicates means ± sem of at least 3 measurements. (B, C, D) Representative images of differential interference contrast (DIC) and fluorescence, and merged images, of cells treated for 1 min at 37°C with Rhodamine-123-loaded nanoparticles (PCL-NP-Rho, 1 µM) (B) or “free” Rhodamine-123 (Rho, 1 µM) (C). Representative images are also shown for cells treated with “empty” PCL-NP's (D).