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Related Experiment Videos

Ca2+-H+ antiport activity in synaptic vesicles isolated from sheep brain cortex

P P Gonçalves1, S M Meireles, C Gravato

  • 1Departamento de Biologia, Universidade de Aveiro, Portugal. pgoncalves@bio.ua.pt

Neuroscience Letters
|July 9, 1998
PubMed
Summary

Sheep brain synaptic vesicles accumulate calcium ions using a proton gradient, driven by an H+-pumping ATPase. This process involves a calcium-hydrogen ion antiporter, crucial for synaptic function.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • Synaptic vesicles are crucial for neurotransmitter release.
  • Calcium ion (Ca2+) homeostasis is vital for synaptic transmission.
  • The mechanisms of Ca2+ transport in synaptic vesicles are not fully elucidated.

Purpose of the Study:

  • To investigate the mechanism of ATP-dependent Ca2+ accumulation in sheep brain synaptic vesicles.
  • To identify the type of ATPase involved in energizing Ca2+ transport.
  • To characterize the relationship between proton gradients and Ca2+ uptake.

Main Methods:

  • Isolation of synaptic vesicles from sheep brain cortex.
  • Measurement of Ca2+ uptake using isotopic methods.
  • Assessment of proton gradient dissipation using acridine orange fluorescence quenching.

Related Experiment Videos

  • Inhibition studies using V-type and P-type ATPase inhibitors (bafilomycin, vanadate) and a protonophore (CCCP).

    • Main Results:

    • Synaptic vesicles exhibit ATP-dependent Ca2+ accumulation.
    • This transport is sensitive to V-type ATPase inhibitors but not P-type ATPase inhibitors.
    • Ca2+ accumulation is linked to proton gradient dissipation and increased ATP hydrolysis.
    • A low-affinity Ca2+-H+ antiport system energized by the H+-pumping ATPase was identified.

    Conclusions:

    • Sheep brain synaptic vesicles utilize a V-type H+-pumping ATPase to establish a proton motive force.
    • This proton motive force energizes a Ca2+-H+ antiport system for Ca2+ accumulation.
    • The findings elucidate a key mechanism for calcium regulation within synaptic vesicles.