Tumor Necrosis Factor Alpha Mediates GABAA Receptor Trafficking to the Plasma Membrane of Spinal Cord Neurons In Vivo
Figure 1
Overview of in vivo nanoinjection methods for confocal imaging experiments. (1) Subjects () received a single TNFα injection dose (0.01, 0.1, or 1 μM) as well as a contralateral control injection of albumin. (2) Injection sites were 1 mm apart along the rostrocaudal axis. Injection sites were localized using FluoroRuby (depicted as red regions). (3) Within injection sites, large ventral motor neurons were selected under wide-field fluorescence in a blinded fashion by using the characteristic pattern of presynaptic synaptophysin outlining the plasma membrane to identify cells. (4) Confocal z-stacks were deblurred by 3D blind iterative deconvolution (AutoQuant). Panels depict a z-stack before (left) and after (right) deconvolution. (5) Custom macros designed using MetaMorph software (Molecular Devices) were used to quantify the number of fluorescently-labeled receptor puncta on the plasma membrane. WM: white matter; GM: gray matter.
