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Immunochemical Analysis of Vesicular Monoamine Transporter (VMAT2) Protein in Parkinson's Disease

https://doi.org/10.1006/exnr.1998.7008Get rights and content

Abstract

The vesicular monoamine transporter (VMAT2) has been suggested to be an excellent marker of presynaptic dopaminergic nerve terminals in the striatum of Parkinson's disease patients based on its high level of expression and insensitivity to drugs used to treat the disease. Previousin vivoimaging and postmortem binding studies have detected a loss in striatal VMAT2 binding in Parkinson's diseased (PD) brain; however, these techniques have poor spatial resolution and may suffer from nonspecific binding of some ligands. In this study, we use novel polyclonal antibodies to distinct regions of human VMAT2 to quantify and localize the protein. Western blot analysis demonstrated marked reductions in VMAT2 immunoreactivity in putamen, caudate, and nucleus accumbens of PD brain compared to control cases. Immunohistochemistry revealed VMAT2 immunoreactive fibers and puncta that were dense throughout the striatum of control brains, but which were drastically reduced in putamen of PD brains. In PD brains the caudate showed a significant degree of sparing along the border of the lateral ventricle and the nucleus accumbens was relatively preserved. The distribution of VMAT2 in striatum and its loss in PD paralleled that of the dopamine transporter (DAT), a phenotypic marker of dopamine neurons. Thus, immunochemical analysis of VMAT2 protein provides novel and sensitive means for localizing and quantifying VMAT2 protein and nigrostriatal dopamine terminals in PD. Furthermore, the relative expression of VMAT2 compared to that of DAT may predict the differential vulnerability of dopamine neurons in PD.

References (54)

  • D.B. Smith et al.

    Single-step purification of polypeptides expressed inEscherichia coliS

    Gene

    (1988)
  • J.K. Staley et al.

    Radioligand binding and immunoautoradiographic evidence for a lack of toxicity to dopaminergic terminals in human cocaine overdose victims

    Brain Res.

    (1997)
  • C.K. Surratt et al.

    A human synaptic vesicle monoamine transporter cDNA predicts post-translational modifications, reveals chromosome 10 gene localization and identifies Taq1 RFLPs

    FEBS Lett.

    (1993)
  • F. Thibaut et al.

    Regional distribution of monoamine vesicular uptake sites in the mesencephalon of control subjects and patients with Parkinson's disease: A postmortem study using tritiated tetrabenazine

    Brain Res.

    (1995)
  • T.M. Vanderborght et al.

    [3

    Neuroscience

    (1995)
  • Y.M. Wang et al.

    Knockout of the vesicular monoamine transporter 2 gene results in neonatal death and hypersensitivity to cocaine and amphetamine

    Neuron

    (1997)
  • V. Blanchard et al.

    Differential expression of tyrosine hydroxylase and membrane dopamine transporter genes in subpopulations of dopaminergic neurons of rat mesencephalon

    Mol. Brain Res.

    (1994)
  • B.J. Ciliax et al.

    The dopamine transporter: Immunochemical characterization and localization in brain

    J. Neurosci.

    (1995)
  • J.N. Dasilva et al.

    In vivo imaging of monoaminergic nerve terminals in normal and MPTP-lesioned primate brain using positron emission tomography (PET) and [11

    Synapse

    (1993)
  • R.H. Edwards

    Neural degeneration and the transport of neurotransmitters

    Ann. Neurol.

    (1993)
  • J.D. Erickson et al.

    Distinct pharmacological properties and distribution in neurons and endocrine cells of two isoforms of the human vesicular monoamine transporter

    Proc. Natl. Acad. Sci. USA

    (1996)
  • J.D. Erickson et al.

    Functional identification and molecular cloning of a human brain vesicle monoamine transporter

    J. Neurochem.

    (1993)
  • J.D. Erickson et al.

    Expression cloning of a reserpine-sensitive vesicular monoamine transporter

    Proc. Natl. Acad. Sci. USA

    (1992)
  • K.A. Frey et al.

    Presynaptic monoaminergic vesicles in Parkinson's disease and normal aging

    Ann. Neurol.

    (1996)
  • Fumagalli, F. R. R. Gainetdinov, Y. M. Wang, G. W. Miller, M. G. Caron, Increased neurotoxicity to methamphetamine in...
  • R.R. Gainetdinov et al.

    Dopamine transporter is required for in vivo MPTP neurotoxicity: Evidence from mice lacking the transporter

    J. Neurochem.

    (1997)
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