CALCIUM REGULATORY MECHANISMS IN NEURAL CELLS

Project: Research project

Description

The project aims to understand the mechanisms that regulate Ca2+ inside
neural cells. Ca2+ fulfills an essential role in neural cells,
controlling key excitatory responses including release of
neurotransmitters, axonal membrane conductance, generation of action
potentials, electrical coupling between neurons, and fast axonal
transport. The proposed studies involve a combination of cell biological
and biochemical approaches to analyze the molecular basis of mechanisms
which control Ca2+ release into the neuronal cytoplasm and transfer Ca2+
inside neural cells. The project centers on Ca2+ transport mechanisms
activated either by inositol phosphates or quanine nucleotides. The
studies have three objectives: (1) Elucidation of the identity and
organization of intracellular Ca2+ pools: studies will assess the
function of Ca2+ regulatory organelles in clonal neuroblastoma cell
cultures and isolated synaptosomes, identify at the subcellular level
those organelles responsive to inositol 1,4,5-trisphophate (InsP3) and
GTP, and use electron microscopy and electron probe microanalysis to
localize intracellular Ca2+ pools within the neural systems. (2)
Determination of the mechanisms of intracellular Ca2+ transport activated
by InsP3 and GTP; measurements will be made on the nature and specificity
of ion transport activated by InsP3, and on the physical and biochemical
basis of the process by which a GTP-regulated mechanism mediates the
translocation of Ca2+ between distinct Ca2+-regulatory compartments within
neuronal cells. (3) Molecular characterization of the Ca2+ translocation
mechanisms; studies are designed to identify the proteins involved in
InsP3- and GTP-activated Ca2+ translocation, and to apply immunochemical
approaches to the determinations of the existence of known Ca2+ -
regulatory proteins within discrete membrane subfractions of neural cells.
The project utilizes a combination of isotopic transport measurements, electron microscopic structural analyses, biophysical measurements on the
interactions between membranes, protein analytical and immunochemical
approaches, aa directed at ascertaining how Ca2+ translocation occurs
within neural cells and what regulates it. In view of the fundamental
role of Ca2+ in controlling neural functional, identification and
characterization of the mechanisms that mediate and regulate Ca2+ signals
in neural cells will permit understanding of the generation of neural
excitability, and the development of methods to control the deleterious
effects of diseases that alter neuronal conduction and excitability.
StatusFinished
Effective start/end date4/1/836/30/95

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $11,237.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $195,713.00
  • National Institutes of Health
  • National Institutes of Health: $183,987.00

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Calcium
Cell Membrane
Guanosine Triphosphate
Neuroblastoma
Organelles
Synaptosomes
Physical Phenomena
Electron Probe Microanalysis
Inositol 1,4,5-Trisphosphate
Inositol Phosphates
Membranes
Ion Transport
Inositol
Electron Transport
Electron Microscopy
Membrane Proteins
Cytoplasm
Proteins
Nucleotides
Cell Line