PMID- 10103130
OWN - NLM
STAT- MEDLINE
DCOM- 19990520
LR  - 20190815
IS  - 0953-816X (Print)
IS  - 0953-816X (Linking)
VI  - 11
IP  - 4
DP  - 1999 Apr
TI  - Unexpected localization of the Na+/Cl--dependent-like orphan transporter, Rxt1,
      on synaptic vesicles in the rat central nervous system.
PG  - 1349-61
AB  - Numerous features of its primary structure demonstrate that the orphan
      transporter Rxt1 belongs to the Na+/Cl--dependent neurotransmitter plasma
      membrane transporter superfamily, which includes the dopamine, norepinephrine,
      serotonin and gamma-aminobutyric acid (GABA) transporters. Initial
      immunocytochemical investigations with affinity-purified antibodies have
      established that Rxt1 is localized, almost exclusively, in axon terminals of
      glutamatergic neurons and subsets of GABAergic neurons in the CNS. Further
      studies were carried out to determine its subcellular distribution. In a first
      series of experiments, PC-12 cells were transfected with plasmids encoding either
      the dopamine transporter or Rxt1. Immunofluorescence experiments showed that the 
      dopamine transporter was expressed in these cells, and, as expected, addressed to
      their plasma membrane. Surprisingly, this was never the case with Rxt1, which was
      targeted to the same subcellular compartment as synaptophysin, a vesicular
      protein. In a second set of experiments, subcellular fractionation of rat
      striatum showed that Rxt1, but not the dopamine transporter, was relatively
      abundant in the purified synaptic vesicle fraction. Finally, electron microscopic
      immunocytochemistry with anti-Rxt1 antibodies showed peroxidase as well as pre-
      and post-embedding immunogold labelling confined to the intracellular compartment
      in various brain regions. Moreover, quantitative analysis of post-embedding
      experiments demonstrated that the immunogold particles corresponding to Rxt1
      immunoreactivity were mostly localized to small synaptic vesicles. These data
      indicate that, in contrast with the other members of the Na+/Cl--dependent
      neurotransmitter transporter superfamily, which are targeted to the plasma
      membrane, Rxt1 is distributed as a vesicular protein in the CNS.
FAU - Masson, J
AU  - Masson J
AD  - INSERM U288, NeuroPsychoPharmacologie Moleculaire, Cellulaire et Fonctionelle,
      Faculte de Medecine, Pitie-Salpetriere, Paris, France.
FAU - Riad, M
AU  - Riad M
FAU - Chaudhry, F
AU  - Chaudhry F
FAU - Darmon, M
AU  - Darmon M
FAU - Aidouni, Z
AU  - Aidouni Z
FAU - Conrath, M
AU  - Conrath M
FAU - Giros, B
AU  - Giros B
FAU - Hamon, M
AU  - Hamon M
FAU - Storm-Mathisen, J
AU  - Storm-Mathisen J
FAU - Descarries, L
AU  - Descarries L
FAU - El Mestikawy, S
AU  - El Mestikawy S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - France
TA  - Eur J Neurosci
JT  - The European journal of neuroscience
JID - 8918110
RN  - 0 (Carrier Proteins)
RN  - 0 (Dopamine Plasma Membrane Transport Proteins)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (Membrane Transport Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Rxt1 protein, rat)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Carrier Proteins/*analysis/metabolism
MH  - Dopamine Plasma Membrane Transport Proteins
MH  - Fluorescent Antibody Technique
MH  - Immunohistochemistry
MH  - Male
MH  - *Membrane Glycoproteins
MH  - *Membrane Transport Proteins
MH  - Microscopy, Electron
MH  - Nerve Tissue Proteins/*analysis/metabolism
MH  - PC12 Cells
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Subcellular Fractions/chemistry
MH  - Synaptic Vesicles/*chemistry
MH  - Transfection
EDAT- 1999/04/02 00:00
MHDA- 1999/04/02 00:01
CRDT- 1999/04/02 00:00
PHST- 1999/04/02 00:00 [pubmed]
PHST- 1999/04/02 00:01 [medline]
PHST- 1999/04/02 00:00 [entrez]
AID - 10.1046/j.1460-9568.1999.00540.x [doi]
PST - ppublish
SO  - Eur J Neurosci. 1999 Apr;11(4):1349-61. doi: 10.1046/j.1460-9568.1999.00540.x.