Figure 3
Post-embedding immunoelectron microscopy for DGKβ in spines of hippocampal neurons. (A-C) Electron micrographs showing immunogold localization of DGKβ in spines of hippocampal neurons. Arrowheads, large arrow, and small arrow indicate perisynaptic, extrasynaptic, or intracellular labeling, respectively. (D) The percentage of immunogold particles for cell membrane-associated (para-plasma membrane distribution) and intracellular (cytoplasmic distribution) labeling in spines of hippocampal neurons (n = 49 immunogold particles in 32 spines). Cell membrane-associated distribution was defined by the position of immunogold particles located less than 35 nm from the cell membrane to the center of gold particles. (E) A histogram showing tangential distribution of cell membrane-associated immunogold particles on spines of hippocampal neurons (n = 37 immunogold particles). DGKβ peaked in the 0–60-nm bin, which is just extrasynaptic from the edge of PSD. Scale bars, 100 nm.