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National Institutes of Health, Bethesda

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

Chan-Ying Zheng¹, Ronald S. Petralia¹, Ya-Xian Wang¹, Bechara Kachar¹

¹National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda

FRAP has been used to quantify the mobility of Green Fluorescence Protein (GFP)-tagged proteins in cultured cells. We examined the mobile/immobile fractions of the GFP by analyzing the fluorescence recovery percentage after photobleaching. In this study, FRAP was performed at spines of hippocampal neurons.

Neuroscience

High-Resolution Quantitative Immunogold Analysis of Membrane Receptors at Retinal Ribbon Synapses

Jun Zhang¹, Ronald S. Petralia², Ya-Xian Wang², Jeffrey S. Diamond¹

¹Synaptic Physiology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, ²Advanced Imaging Core, National Institute on Deafness and Other Communication Disorders, National Institutes of Health

The postembedding immunogold method is one of the most effective ways to provide high-resolution analyses of the subcellular localization of specific molecules. Here we describe a protocol to quantitatively analyze glutamate receptors at retinal ribbon synapses.