The dopamine transporter is absent in Parkinsonian putamen and reduced in the caudate nucleus

Hyman B. Niznik, Evan Fogel, Frank F. Fassos, Philip Seeman

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

The neuronal dopamine transporter/uptake site can be covalently labeled with the photoaffinity probe 1-(2 -[bis -(4-fluorophenyl)methoxy]ethyl)-4-[2-(4- azido-3-[125I]iodophenyl)ethyl]piperazine ([125I]FAPP) and visualized following sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Upon photolysis, [125I]FAPP specifically incorporated into a polypeptide of apparent Mr = 62,000 in membranes from both the putamen and the caudate nucleus of control, Alzheimer's, schizophrenia, and Huntington's diseased brain, and, following complete deglycosylation, migrated as an Mr ∼48,000 polypeptide. In parkinsonian postmortem putamen, however, there was no detectable photoincorporation of [125I]FAPP into the ligand binding subunit of the dopamine transporter. [125I]FAPP did specifically label the Mr 62,000 polypeptide of parkinsonian caudate, although with efficiencies of 20-50% of control. The asymmetrical loss of the dopamine transporter in Parkinson's diseased striatum was confirmed in reversible receptor binding experiments using [3H]GBR-12935 {3H-labeled 1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine}. In parkinsonian putamen, mazindol competitively inhibited the binding of [3H]GBR-12935 with an estimated affinity (Ki, ∼ 2,000 nM) 10 times lower than in controls (Ki, ∼ 30 nM). while the affinity of mazindol for [3H]GBR-12935 binding in the caudate was equal to that seen with controls (Ki ∼ 50 nM). The proportion of [3H]GBR-12935 binding sites recognized by mazindol with high affinity in Parkinson's diseased caudate was, however, reduced by 50-80%. Taken together, these data clearly suggest that (a) the dopamine transporter is not detectable in Parkinson's diseased putamen, (b) [3H]GBR-12935 binding in the putamen of Parkinson's diseased brain is probably not to the dopamine uptake carrier, and (c) there is an interregional variation in the loss of dopamine uptake sites (lost in putamen and somewhat spared in caudate), which appears to parallel markedly the reported loss of dopamine content in idiopathic Parkinson's diseased striatum.

Original languageEnglish (US)
Pages (from-to)192-198
Number of pages7
JournalJournal of Neurochemistry
Volume56
Issue number1
StatePublished - 1991
Externally publishedYes

Fingerprint

Dopamine Plasma Membrane Transport Proteins
Caudate Nucleus
Putamen
Mazindol
Parkinson Disease
Dopamine
Peptides
Brain
Photolysis
Huntington Disease
Electrophoresis
Autoradiography
Sodium Dodecyl Sulfate
GBR 12935
Labels
Polyacrylamide Gel Electrophoresis
Schizophrenia
Alzheimer Disease
Binding Sites
1-(2-(bis-(4-fluorophenyl)methoxy)ethyl)-4-(2-(4-azido-3-iodophenyl)ethyl)piperazine

Keywords

  • 12935
  • [H]GBR
  • Dopamine uptake
  • Parkinson's disease
  • Photoaffinity labeling

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

The dopamine transporter is absent in Parkinsonian putamen and reduced in the caudate nucleus. / Niznik, Hyman B.; Fogel, Evan; Fassos, Frank F.; Seeman, Philip.

In: Journal of Neurochemistry, Vol. 56, No. 1, 1991, p. 192-198.

Research output: Contribution to journalArticle

Niznik, Hyman B. ; Fogel, Evan ; Fassos, Frank F. ; Seeman, Philip. / The dopamine transporter is absent in Parkinsonian putamen and reduced in the caudate nucleus. In: Journal of Neurochemistry. 1991 ; Vol. 56, No. 1. pp. 192-198.
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N2 - The neuronal dopamine transporter/uptake site can be covalently labeled with the photoaffinity probe 1-(2 -[bis -(4-fluorophenyl)methoxy]ethyl)-4-[2-(4- azido-3-[125I]iodophenyl)ethyl]piperazine ([125I]FAPP) and visualized following sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Upon photolysis, [125I]FAPP specifically incorporated into a polypeptide of apparent Mr = 62,000 in membranes from both the putamen and the caudate nucleus of control, Alzheimer's, schizophrenia, and Huntington's diseased brain, and, following complete deglycosylation, migrated as an Mr ∼48,000 polypeptide. In parkinsonian postmortem putamen, however, there was no detectable photoincorporation of [125I]FAPP into the ligand binding subunit of the dopamine transporter. [125I]FAPP did specifically label the Mr 62,000 polypeptide of parkinsonian caudate, although with efficiencies of 20-50% of control. The asymmetrical loss of the dopamine transporter in Parkinson's diseased striatum was confirmed in reversible receptor binding experiments using [3H]GBR-12935 {3H-labeled 1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine}. In parkinsonian putamen, mazindol competitively inhibited the binding of [3H]GBR-12935 with an estimated affinity (Ki, ∼ 2,000 nM) 10 times lower than in controls (Ki, ∼ 30 nM). while the affinity of mazindol for [3H]GBR-12935 binding in the caudate was equal to that seen with controls (Ki ∼ 50 nM). The proportion of [3H]GBR-12935 binding sites recognized by mazindol with high affinity in Parkinson's diseased caudate was, however, reduced by 50-80%. Taken together, these data clearly suggest that (a) the dopamine transporter is not detectable in Parkinson's diseased putamen, (b) [3H]GBR-12935 binding in the putamen of Parkinson's diseased brain is probably not to the dopamine uptake carrier, and (c) there is an interregional variation in the loss of dopamine uptake sites (lost in putamen and somewhat spared in caudate), which appears to parallel markedly the reported loss of dopamine content in idiopathic Parkinson's diseased striatum.

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