Expression cloning of a cocaine-and antidepressant-sensitive human noradrenaline transporter

  title={Expression cloning of a cocaine-and antidepressant-sensitive human noradrenaline transporter},
  author={Tadeusz Pacholczyk and Randy D. Blakely and Susan Gaye Amara},
AT most synapses, chemical signalling is terminated by a rapid reaccumulation of neurotransmitter into presynaptic terminals1–5. Uptake systems for the biogenic amines are the initial site of action for therapeutic antidepressants and drugs such as cocaine and the amphetamines. We have isolated a complementary DNA clone encoding a human noradrenaline transporter. The cDNA sequence predicts a protein of 617 amino acids, with 12–13 highly hydrophobic regions compatible with membrane-spanning… 

Cloning and expression of a functional serotonin transporter from rat brain

A large family of related gene products expressed in rodent brain is identified from two highly conserved regions of the transporters for noradrenaline and γ-aminobutyric acid, and one of these products hybridizes to a single 3.7-kilobase RNA restricted to rat midbrain and brainstem, where it is highly enriched within the serotonergic raphe complex.

Molecular physiology of norepinephrine and serotonin transporters.

Electrophysiological recording of human NETs and SERTs stably expressed in HEK-293 cells reveals that both transporters move charge across the plasma membrane following the addition of substrates; these currents can be blocked by NET-and SERT-selective antagonists as well as by cocaine.

Molecular Biology of Serotonin Uptake Sites

Using an expression cloning strategy, a cDNA for an antidepressant-sensitive, Nat-dependent 5-HT reuptake protein from rat mast cells is isolated and the cDNA predicts a 630-amino acid protein with 12 putative transmembrane domains and extensive homology to GABA, norepinephrine (NE), and dopamine (DA) transporters.

Immunolocalization of the cocaine‐ and antidepressant‐sensitive l‐norepinephrine transporter

Initial ultrastructural evaluation via preembedding immunogold techniques revealed substantial cytoplasmic NET immunoreactivity in axon terminals within the prelimbic prefrontal cortex, consistent with postulates of regulated trafficking controlling neurotransmitter clearance.

Identification and characterization of antidepressant-sensitive serotonin transporter proteins using site-specific antibodies

These studies indicate that brain and platelet SERTs are formed from identical polypeptides differing significantly in their extent of N- linked glycosylation, and that SERT- immunoreactive fibers were found to be widely distributed throughout the rodent brain.

Murine serotonin transporter: sequence and localization to Chromosome 11

The cDNA and deduced amino acid sequence of the mouse SERT is described, comparison with the rat SERT, the chromosomal localization of themouse SERT gene, and the predicted location of the homologous human gene are described.

[Pharmacology of monoamine neurotransmitter transporters].

The molecular cloning of these transporters revealed that NET, DAT, and SERT are members of a sodium-dependent neurotransmitter transporter geneFamily, while VMATs arise from proton-dependent transporter gene family, suggesting both activity-dependent and pharmacological regulatory mechanisms for transporter expression.

A cDNA that suppresses MPP+ toxicity encodes a vesicular amine transporter

A cocaine insensitive chimeric insect serotonin transporter reveals domains critical for cocaine interaction.

Results show TMD1 and TMD2 affect the apparent substrate transport and antagonist sensitivity by possibly providing unique conformations to the transporter and the availability of these chimeras facilitates elucidation of specific amino acids involved in interactions with cocaine.



Cloning and expression of a rat brain GABA transporter.

Hydropathy analysis of the deduced protein suggests multiple transmembrane regions, a feature shared by several cloned transporters; however, database searches indicate that GAT-1 is not homologous to any previously identified proteins.

Dopamine transporter: biochemistry, pharmacology and imaging.

In vivo labeling studies of the dopamine transporter have revealed the usefulness of imaging the transporter, a measure of the presence of dopaminergic nerve terminals, as a potential diagnostic tool in Parkinson's disease.

Cocaine receptors on dopamine transporters are related to self-administration of cocaine.

It is shown here that the potencies of cocaine-like drugs in self-administration studies correlate with their potencies in inhibiting [3H]mazindol binding to the dopamine transporters in the rat striatum, but not with theirPotencies in binding to a large number of other presynaptic and postsynaptic binding sites.

High‐Affinity Uptake of [3H]Norepinephrine by Primary Astrocyte Cultures and Its Inhibition by Tricyclic Antidepressants

These characteristics are essentially identical to those found for high‐affinity uptake of NE in total membrane or synaptosome fractions from rodent brains and suggests that such uptake in neural tissue is not exclusively neuronal.

Mechanism of Transport and Storage of Neurotransmitter

This review will focus on the bioenergetics, mechanism, and molecular basis of neurotransmitter transport, and the sodium-coupled and proton coupled systems, which play an important role in the overall process of synaptic transmission.

Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-1,2,3,6 -tetrahydropyridine: uptake of the metabolite N-methyl-4-phenylpyridine by dopamine neurons explains selective toxicity.

Blockage of dopamine uptake by mazindol prevents MPTP-induced damage to nigrostriatal dopamine neurons, indicating that MPP+ concentration into dopamine neurons explains their selective destruction by MPTP.