Cellular resistance to Evans blue toxicity involves an up‐regulation of a phosphate transporter implicated in vesicular glutamate storage

It has recently been suggested that the brain‐specific Na+‐dependent phosphate inorganic co‐transporter (BNPI) is able to support glutamate transport and storage in synaptic vesicles. A procedure for measuring the vesicular pool of glutamate is described and was used to select cell lines according t...

Full description

Saved in:
Bibliographic Details
Published in:Journal of neurochemistry 2001-08, Vol.78 (3), p.658-663
Main Authors: Israël, M., Tomasi, M., Bostel, S., Meunier, F.‐M.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Carrier Proteins
Carrier Proteins - genetics
Carrier Proteins - metabolism
Cell Line
Cell physiology
Cerebellum
Cerebellum - cytology
Coloring Agents
Coloring Agents - toxicity
Drug Resistance
Drug Resistance - physiology
Evans Blue
Evans Blue - toxicity
Fundamental and applied biological sciences. Psychology
Glutamic Acid
Glutamic Acid - chemistry
Glutamic Acid - metabolism
inorganic phosphate transporter
Life Sciences
Molecular and cellular biology
Neurons and Cognition
Neurotransmission
phosphate transporter
Protein Isoforms
Reverse Transcriptase Polymerase Chain Reaction
Sodium-Phosphate Cotransporter Proteins
Symporters
Synaptosomes
Synaptosomes - metabolism
vesicular glutamate
vesicular transporter
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:It has recently been suggested that the brain‐specific Na+‐dependent phosphate inorganic co‐transporter (BNPI) is able to support glutamate transport and storage in synaptic vesicles. A procedure for measuring the vesicular pool of glutamate is described and was used to select cell lines according to their glutamate storage capacity. Two cell lines were selected: C6BU‐1, with a large intracellular glutamate storage capacity, and NG108‐15, devoid of it. Their contents in BNPI mRNA were compared by RT‐PCR. We found that both cell lines had BNPI, but in addition C6BU‐1 alone expresses the other isoform, DNPI. We also carried out a clonal selection of NG108‐15 cells in the presence of the dye Evans blue, a competitive inhibitor of vesicular glutamate transport, very toxic for cells in culture. It was assumed that only those that sequester and eliminate the drug by overexpressing a vesicular glutamate transporter would survive. We found that the NG108‐15 clones resistant to Evans blue had an increased storage capacity for glutamate. These cells also up‐regulated the BNPI isoform of the phosphate transporter as shown by RT‐PCR and northern blot.
ISSN:0022-3042
1471-4159
DOI:10.1046/j.1471-4159.2001.00449.x