title: Fluorescence methods for analysis of interactions between Ca(2+) signaling, lysosomes, and endoplasmic reticulum.
abstract: The endoplasmic reticulum (ER) is both the major source of intracellular Ca(2+) for cell signaling and the organelle that forms the most extensive contacts with the plasma membrane and other organelles. Lysosomes fulfill important roles in degrading cellular materials and in cholesterol handling, but they also contribute to Ca(2+) signaling by both releasing and sequestering Ca(2+). Interactions between ER and other Ca(2+)-transporting membranes, notably mitochondria and the plasma membrane, often occur at sites where the two membranes are closely apposed, allowing local Ca(2+) signaling between them. These interactions are often facilitated by scaffold proteins. Recent evidence suggests similar local interactions between ER and lysosomes. We describe simple fluorescence-based methods that allow the interplay between Ca(2+) signals, the ER, and lysosomes to be examined.
authors: Department of Pharmacology, University of Cambridge, Cambridge, UK.
authors: David L
authors: DL
authors: Prole
authors: Department of Pharmacology, University of Cambridge, Cambridge, UK.
authors: Cristina I
authors: CI
authors: López-Sanjurjo
authors: Department of Pharmacology, University of Cambridge, Cambridge, UK.
authors: Stephen C
authors: SC
authors: Tovey
authors: Department of Pharmacology, University of Cambridge, Cambridge, UK.
authors: Colin W
authors: CW
authors: Taylor
mesh_terms: D015220:Calcium Channels / Q000737:chemistry* / Q000502:physiology; D020013:Calcium Signaling; D002463:Cell Membrane Permeability; D004721:Endoplasmic Reticulum / Q000502:physiology*; D057809:HEK293 Cells; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D008247:Lysosomes / Q000502:physiology*; D008564:Membrane Potentials; D018408:Patch-Clamp Techniques; D000097903:Two-Pore Channels
chemical_list: D015220:Calcium Channels; D008051:Lipid Bilayers; C407357:TPCN1 protein, human; D000097903:Two-Pore Channels; C530224:TPCN2 protein, human
keywords: Ca(2+); Endoplasmic reticulum; Fluorescence; Fluorescent proteins; Inositol trisphosphate receptor; Lysosomes; Optical microscopy