Many receptor-mediated endocytic processes are mediated by constitutive budding of clathrin-coated pits (CCPs) at spatially randomized sites before slowly pinching off from the plasma membrane (60-100 s). In contrast, clathrin-mediated endocytosis (CME) coupled with regulated exocytosis in excitable cells occurs at peri-exocytic sites shortly after vesicle fusion (∼10 s). The molecular mechanis...

Accurately detecting and classifying vesicle-plasma membrane fusion events in fluorescence microscopy, is of primary interest for studying biological activities in a close proximity to the plasma membrane. In this paper, we present a novel Gaussian mixture model for automated identification of glucose transporter 4 (GLUT4) vesicle and plasma membrane fusions and classification of full fusion an...

Presynaptic terminals contain several specialized compartments, which have been described by electron microscopy. We show in an identified Drosophila neuromuscular synapse that several of these compartments-synaptic vesicle clusters, presynaptic plasma membrane, presynaptic cytosol, and axonal cytoskeleton-labeled by specific reagents may be resolved from one another by laser scanning confocal ...

Docking to the plasma membrane prepares vesicles for rapid release. Here, we describe a mechanism for dense core vesicle docking in neurons. In Caenorhabditis elegans motor neurons, dense core vesicles dock at the plasma membrane but are excluded from active zones at synapses. We have found that the calcium-activated protein for secretion (CAPS) protein is required for dense core vesicle dockin...

Chemical neurotransmission is the major mechanism of neuronal communication. Neurotransmitters are released from secretory organelles, synaptic vesicles (SVs) via exocytosis into cleft. Fusion SVs with presynaptic plasma membrane balanced by endocytosis, thus maintaining at steady-state levels. The protein machineries responsible for exo- and endocytosis have been extensively investigated. In c...

In the Fluid Mosaic Model for biological membrane structure, proposed by Singer and Nicolson in 1972, the lipid bilayer is represented as a neutral two-dimensional solvent in which the proteins of the membrane are dispersed and distributed randomly. The model portrays the membrane as dominated by a membrane lipid bilayer, directly exposed to the aqueous environment, and only occasionally interr...

Insulin-stimulated glucose uptake requires the activation of several signaling pathways to mediate the translocation and fusion of GLUT4 vesicles to the plasma membrane. Our previous studies demonstrated that GLUT4-mediated glucose uptake is a myosin II-dependent process in adipocytes. The experiments described in this report are the first to show a dual role for the myosin IIA isoform specific...

Evoked exocytosis in excitable cells is fast and spatially confined and must be followed by coupled endocytosis to enable sustained exocytosis while maintaining the balance of the vesicle pool and the plasma membrane. Various types of exocytosis and endocytosis exist in these excitable cells, as those has been found from different types of experiments conducted in different cell types. Correlat...

To evaluate the possible role of a putative vesicle-targeting protein, syntaxin-4, in vasopressin-regulated trafficking of aquaporin-2 water channel vesicles to the apical plasma membrane of renal collecting duct cells, we have carried out immunoblotting, immunocytochemistry, and reverse transcription (RT)-PCR experiments in rat kidney. Immunochemical studies used an affinity-purified, peptide-...

Transport of newly synthesized cholesterol and vesicular stomatitis virus G protein from the endoplasmic reticulum to the plasma membrane is interrupted by incubation at 15 degrees C. Under this condition the newly synthesized molecules accumulate in both the endoplasmic reticulum (ER) and a subcellular vesicle fraction of low density called the lipid-rich vesicle fraction. The material in the ...