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J O Schenk

Publications and source records attributed to J O Schenk.

At least 19 recordsLinked to original sources
PubMed2001

Characterization of extracellular dopamine clearance in the medial prefrontal cortex: role of monoamine uptake and monoamine oxidase inhibition.

In vitro rotating disk electrode (RDE) voltammetry and in vivo microdialysis were used to characterize dopamine clearance in the rat medial prefrontal cortex (mPFC). RDE studies indicate that inhibition by cocaine, specific inhibitors of the dopamine transporter (DAT) and norepinephrine transporter (NET), and low Na(+) produced a 50-70% decrease in the velocity of dopamine clearance. Addition of the monoamine (MAO) inhibitors, l-deprenyl, clorgyline, pargyline, or in vivo nialamide produced 30-50% inhibition. Combined effects of uptake inhibitors with l-deprenyl on dopamine clearance were additive (up to 99% inhibition), suggesting that at least two mechanisms may contribute to dopamine clearance. Dopamine measured extracellularly 5 min after exogenous dopamine addition to incubation mixtures revealed that most conditions of DAT/NET inhibition did not produce elevated dopamine levels above controls. Inhibition of MAO produced elevated dopamine levels only after long-term, but not short-term, incubation in vitro. Short-term incubation of l-deprenyl combined with DAT and NET uptake inhibitors increased dopamine above control levels, consistent with more than one mechanism of dopamine clearance. Local infusion of pargyline (100 or 300 microm) into the mPFC or striatum via microdialysis produced more pronounced and immediate increases in mPFC dopamine levels compared with striatum. Furthermore, dopamine elevation in the mPFC was not accompanied by a decrease in the dopamine metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, as found in the striatum. These findings may have revealed a unique mechanism of mPFC dopamine clearance and therefore contribute to the understanding of multiple behaviors that involve mPFC dopamine transmission, such as schizophrenia, drug abuse, and working memory function.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1999

Multisubtrate mechanism for the inward transport of dopamine by the human dopamine transporter expressed in HEK cells and its inhibition by cocaine.

Rotating disk electrode voltammetry was used to measure the time-resolved inward transport of dopamine into human embryonic kidney cells expressing the human transporter for dopamine and a kinetic mechanism of transport is hypothesized. Dopamine transport in this preparation was highly concentrative, with a 10(6)-10(7) inward bias, first order in dopamine and the K(m) and V(max) were found to be 1.6 microM and 18 pmol/sec x 10(6) cells), respectively. The hDAT turnover was estimated to be approximately 18 s(-1) and the second order rate constant of association of dopamine with hDAT was approximately 10(7) M(-1)s(-1). Dopamine transport was found to have a second order dependence on Na(+) (K(Na) approximately 100 mM) and a first order dependence on Cl(-) (K(Cl) approximately 12 mM). Multisubstrate analyses suggested that hDAT operates with an ordered kinetic mechanism in which Na(+) binds first to the transporter protein, dopamine second, and Cl(-) last before translocation of dopamine into or across the membrane. Cocaine competitively inhibited dopamine transport (reaction order of unity and K(i) approximately 0.34 microM) with no discernible effect at the Na(+) and Cl(-) binding sites. These results differ from those of previous studies conducted in preparations of the striatum and nucleus accumbens. Comparisons of the variant results are made and an analysis of the differing apparent kinetic mechanisms is presented.

Carrier Proteins↗
PubMed1999

Effects of methylphenidate analogues on phenethylamine substrates for the striatal dopamine transporter: potential as amphetamine antagonists?

Methylphenidate (MPD) was found to inhibit competitively the striatal dopamine transporter (DAT) and bind at sites on the DAT in common with both cocaine (a non-substrate site ligand) and amphetamine (a substrate site ligand). Some methylphenidate analogues modified on the aromatic ring and/or at the nitrogen were tested to determine whether the profile of inhibition could be altered. None was found to stimulate the release of dopamine in the time frame (< or = 60 s) of the experiments conducted, and each of the analogues tested was found to noncompetitively inhibit the transport of dopamine. It was found that halogenating the aromatic ring with chlorine (threo-3,4-dichloromethylphenidate hydrochloride; compound 1) increased the affinity of MPD to inhibit the transport of dopamine. A derivative of MPD with simultaneous, single methyl group substitutions on the phenyl ring and at the nitrogen (threo-N-methyl-4-methylphenidate hydrochloride; compound 2) bound at a site in common with MPD. A benzyl group positioned at the nitrogen (threo-N-benzylmethylphenidate hydrochloride; compound 3) imparted properties to the inhibitor in which binding at substrate and non-substrate sites could be distinguished. This analogue bound at a mutually interacting site with that of methylphenidate and had a K(int) value of 4.29 microM. Furthermore, the N-substituted analogues (compounds 2 and 3), although clearly inhibitors of dopamine transport, were found to attenuate dramatically the inhibition of dopamine transport by amphetamine, suggesting that the development of an antagonist for substrate analogue drugs of abuse may be possible.

Algorithms↗
PubMed1998

Protein kinase A activity may kinetically upregulate the striatal transporter for dopamine.

The neuronal dopamine transporter (DAT) plays a key role in terminating dopaminergic chemical neurotransmission; thus, the study of the regulation of DAT activity is important in defining parameters relevant to the control of dopaminergic neurotransmission. Interpretation of the results from previous work of this laboratory suggests that occupation of presynaptic autoreceptors increases DAT activity. Second messenger signaling related to kinetic upregulation of DAT has not been examined previously. However, others have shown that protein kinase C activity may downregulate DAT activity, whereas protein kinase A has shown variable results. Herein it is shown that protein kinase A activity mediates the kinetic upregulation of DAT. Quinpirole increased DAT activity that was blocked by sulpiride and the protein kinase A selective inhibitor H-89. Brief incubations with forskolin and 8-bromo-cAMP (8-Br-cAMP) were found to stimulate striatal DAT activity by increasing the Vmax of transport without affecting the Km. Exposures >15 min had no effect. The 8-Br-cAMP-stimulated increases in DAT activity were blocked by pre-exposure to H-89. Thus, second messenger signaling via the cAMP cascade may mediate kinetic upregulation of DAT. Kinetic analyses of the results suggest that either insertion of DAT into the membrane or activation of pre-existing DAT within the membrane mediates the regulation.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1998

Rotating disk electrode voltammetric measurements of dopamine transporter activity: an analytical evaluation.

Rotating disk electrode (RDE) voltammetry at glassy carbon electrodes in 300- to 500-microL volumes has been shown in the literature by this laboratory and others to be useful in measuring dopamine and norepinephrine inward transport and release in preparations from rat brain and in expression systems. However, an analytical evaluation of the technique has not been made, and it is presented here in studies in striatal suspensions and human embryonic kidney cells expressing the human transporter for dopamine. The RDE was found to be able to measure dopamine and its metabolites with linear responses over the range of physiologically relevant concentrations with practical limits of quantification in the 10 to 50 nM range without signal and/or signal to noise enhancements. Response times of the technique were found to vary between 20 and 60 ms depending on rotation rate. Release and inward transport velocities were shown to be kinetically resolved. The glassy carbon electrode was found to be useful for several years at physiological temperatures without significant changes in electrochemical surface area, residual current, or response factors. Some sources of error in the measurement of release and inward transport values of velocity were noted and described.

Animals↗
PubMed1998

Relationships between the catechol substrate binding site and amphetamine, cocaine, and mazindol binding sites in a kinetic model of the striatal transporter of dopamine in vitro.

Experiments were conducted to determine how (-)-cocaine and S(+)-amphetamine binding sites relate to each other and to the catechol substrate site on the striatal dopamine transporter (sDAT). In controls, m-tyramine and S(+)-amphetamine caused release of dopamine from intracellular stores at concentrations > or = 12-fold those observed to inhibit inwardly directed sDAT activity for dopamine. In preparations from animals pretreated with reserpine, m-tyramine and S(+)-amphetamine caused release of preloaded dopamine at concentrations similar to those that inhibit inwardly directed sDAT activity. S(+)-Amphetamine and m-tyramine inhibited sDAT activity for dopamine by competing for a common binding site with dopamine and each other, suggesting that phenethylamines are substrate analogues at the plasmalemmal sDAT. (-)-Cocaine inhibited sDAT at a site separate from that for substrate analogues. This site is mutually interactive with the substrate site (K(int) = 583 nM). Mazindol competitively inhibited sDAT at the substrate analogue binding site. The results with (-)-cocaine suggest that the (-)-cocaine binding site on sDAT is distinct from that of hydroxyphenethylamine substrates, reinforcing the notion that an antagonist for (-)-cocaine binding may be developed to block (-)-cocaine binding with minimal effects on dopamine transporter activity. However, a strategy of how to antagonize drugs of abuse acting as substrate analogues is still elusive.

Amphetamine↗
PubMed1998

Food deprivation decreases mRNA and activity of the rat dopamine transporter.

We have hypothesized that the midbrain dopamine (DA) neurons are a target for insulin action in the central nervous system (CNS). In support of this hypothesis, we have previously demonstrated that direct intracerebroventricular infusion of insulin results in an increase in mRNA levels for the DA reuptake transporter (DAT). In this study, 24- to 36-hour food deprivation was used as a model of decreased CNS insulin levels, to test whether DAT mRNA levels, DAT protein concentration or DAT functional activity would be decreased. DAT mRNA levels, assessed by in situ hybridization, were significantly decreased in the ventral tegmental area/substantia nigra pars compacta (VTA/SNc) (77 +/- 7% of controls, p < 0.05) of food-deprived (hypoinsulinemic) rats. Binding of a specific high-affinity DAT ligand (125I-RTI-121) to membranes from brain regions of fasted or free-feeding rats provided an estimate of DAT protein, which was unchanged in both of the major terminal projection fields, the striatum and nucleus accumbens (NAc). In addition, we utilized the rotating disk electrode voltametry technique to assess possible changes in the function of the DAT in fasting (hypoinsulinemic) rats. The Vmax of DA uptake was significantly decreased (87 +/- 7% of control, p < 0.05), without a change in the Km of uptake, in striatum from fasted rats. In vitro incubation with a physiological concentration (1 nM) of insulin resulted in an increase of striatal DA uptake to control levels. We conclude that striatal DAT function can be modulated by fasting and nutritional status, with a contribution by insulin.

Animals↗
PubMed1997

Repeated cocaine and stress increase dopamine clearance in the rat medial prefrontal cortex.

The effects of repeated footshock stress or cocaine on the kinetics of dopamine clearance in the medial prefrontal cortex (mPFC) were measured by rotating disk electrode voltammetry (RDEV). Five groups of rats were used: animals were either naive (non-handled), pre-treated with five daily saline (1 ml/kg i.p.) or cocaine (15 mg/kg i.p.) injections, or pre-treated with five daily 20-min sessions of sham shock or footshock (0.05 mA/200 ms/s). Dopamine clearance was measured after a 1-week withdrawal period. No difference in Km values was present among the treatment groups, with the mean Km value at approximately 0.5 microM for all groups. However, Vmax values were approximately 50% higher in daily sham shock-, footshock- and cocaine-pre-treated animals compared to naive rats. The increased ability to remove dopamine in these animals suggests that altered dopamine clearance may serve an adaptive mechanism in the mPFC.

Animals↗
PubMed1997

A multisubstrate kinetic mechanism of dopamine transport in the nucleus accumbens and its inhibition by cocaine.

Kinetic studies of dopamine transport into suspensions of nucleus accumbens (NAcc) and effects of Na+ and Cl- as cosubstrates were performed using rotating disk electrode voltammetry. To mimic chemical neurotransmission, dopamine was added as a rapid pulse, and transporter-mediated clearance of dopamine was evaluated kinetically. This paradigm was shown to approximate a zero trans entry transport experiment. Dopamine was taken up with apparent Km and Vmax values of 1.3 microM and 375 pmol/s/g wet weight, respectively. Transport exhibited apparent trans acceleration. Substitution of Na+ with choline or Cl- with isethionate reduced dopamine transport with reaction orders of two and unity, respectively, accompanied by reductions in Vmax with no changes in Km. Apparent K(Na) and K(Cl) values were 70.0 and 92.1 mM, respectively. Dopamine transport in NAcc was found to follow a partially random, sequential mechanism in which dopamine and Na+ bind randomly to the transporter followed by binding of Cl- before transport. Cocaine inhibited dopamine transport and the influences of the other substrates allosterically with an overall Ki of 0.30 microM. Thus, the general kinetic mechanism of the transport of dopamine in the NAcc is identical to that previously reported by this laboratory for dopamine transport in the striatum. However, the dopamine transporter in the NAcc is more tightly regulated by Na+, possesses a higher kinetic turnover rate, is four times more sensitive to cocaine than the striatal transporter, and exhibits cocaine inhibition independent of [substrate]. These findings suggest that cocaine modulates chemical signaling in NAcc differently than in striatum, providing down-regulation of function irrespective of [substrate], thereby enhancing dopaminergic signaling more robustly in the NAcc than in the striatum.

Animals↗
PubMed1994

Effects of cocaine and repeated cocaine followed by withdrawal. Alterations of dopaminergic transporter turnover with no changes in kinetics of substrate recognition.

The turnover of the transporter for dopamine (ca. 1.5 sec-1) and the apparent second order rate constant of association of dopamine with the outward facing form of the transporter protein (ca. 10(6) M-1sec-1) were estimated using kinetic data from rotating disk voltammetric measures of the inward transport of dopamine in striatal suspensions, standard treatments of the kinetics of transport, and values in the literature for density of striatal transporter sites. Under apparent steady-state conditions of transporter functioning, inhibition of the transport of dopamine by cocaine following its addition to the incubation buffer was found to decrease the turnover of the transporter and not affect the kinetics of substrate recognition. The kinetics of binding of dopamine to the transporter were estimated also by apparent pre-steady-state kinetics. These experiments confirmed the second order nature of the binding of dopamine to the transporter and the numerical value of the rate constant estimated under steady-state conditions; they also demonstrated that the binding of dopamine has an absolute dependence on Na+, and that the second order rate constant of association of dopamine with its transporter is not influenced by cocaine. In separate studies, similar experiments were conducted in tissues from animals that had been treated with cocaine for 3 days and withdrawn for 1 day or 2 weeks. Repeated treatments with cocaine followed by either a 24-hr or 2-week period of withdrawal resulted in increases in the Vmax and turnover of the transporter with no apparent changes in the kinetics of association of substrate. No differences between the Ki for cocaine observed in direct inhibitions of the transport of dopamine and the Ki for cocaine observed in tissues obtained from animals treated repeatedly with cocaine were observed. Taken together, these data suggest that cocaine exerts its effects by altering an intramembrane translocation step for the movement of dopamine and not by changing the recognition of dopamine by the externally facing binding site or the apparent Ki for cocaine. Finally, repeated treatments with cocaine followed by a period of withdrawal appear to kinetically activate the transporter for dopamine.

Animals↗
PubMed1994

Striatal transporter for dopamine: catechol structure-activity studies and susceptibility to chemical modification.

The apparent second-order association rate constant of dopamine binding to the striatal transporter (approximately 1 x 10(6) M-1 s-1) as well as the transporter turnover number (approximately 1.5 s-1) was estimated using rotating disk electrode voltammetry to monitor apparent zero trans entry of dopamine into striatal suspensions. The substrate specificity of the transporter was also assessed using catechol derivatives. Dopamine and norepinephrine were transported, whereas epinephrine and the acidic metabolites of dopamine were not transported. The metabolite, 3-methoxytyramine, was transported with a Km seven times greater than and a Vmax close to that for dopamine. 4-Methoxytyramine was transported more facilely than the 3-methoxy derivative. N-Alkylation of the amine side chain of dopamine reduced transport dramatically. 4-Ethylcatechol and 3,4-dihydroxybenzylamine were transported with velocities 79 and 91% less than that for dopamine, respectively. The rigid analogue 6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene was transported with a greater velocity than the 5,7-dihydroxy derivative. Finally, the apparent Km values for 4-ethylcatechol, 1-amino-2-phenylethane, tyramine, and m-tyramine as cosubstrates with dopamine were 1.1, 11, 17, and 2.6 microM, respectively. Pretreatments of striatal suspensions with chloroethylnorapomorphine, N-ethylmaleimide, Hg2+, 4,5-dihydroxy-4,5-dioxo-1H-pyrrolo[2,3-f] quinoline-2,7,9-tricarboxylic acid (a redox modulator of receptors in neuronal as well as other tissues), and neuraminidase reduced the velocity of transport of dopamine, whereas N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline had no effect. Thus, the dopamine transporter requires an intact catechol with a primary ethylamine side chain for optimal activity relative to shorter side chain derivatives (side chains longer than two carbons were not tested), the 3-hydroxyl group of dopamine is the more critical hydroxyl group, and the beta rotamer of the extended conformation of dopamine is transported preferentially. The catechol appears to mediate the recognition of the substrate, whereas the amine side chain apparently facilitates the conformational change of the transporter that results in movement of dopamine into or across the membrane.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

The striatal transporter for dopamine in the rat may be kinetically up-regulated following 3 weeks of withdrawal from cocaine self-administration.

Rotating disk electrode voltammetry was used to measure the inwardly directed Vmax and Km of dopamine with its transporter in striatal suspensions prepared from nonhandled control rats, rats that had been trained to self-administer cocaine for 20 days (at 26 mg/day per rat) via a jugular catheter and subsequently withdrawn for 3 weeks, and rats that had received saline (155 mM NaCl) via a jugular catheter on the same schedule as the rats that had received cocaine. Because a limited number of animals was available from the self-administration procedure, the velocity of dopamine transport as a function of [dopamine] was measured by incremental addition of dopamine to a given striatal preparation. In nonhandled controls the values of Vmax, Km, and turnover, observed in this experimental paradigm, were increased relative to results obtained in studies of the velocity-[dopamine] relationship where dopamine was added to suspensions, one concentration per suspension. The kinetics of the association of dopamine with the transporter were unchanged. The Vmax to Km ratios obtained in the two experiments were statistically indistinguishable, suggesting that the two types of experiments probe the same transporter. Also, the increased velocity observed in the experiment involving sequential additions to the same preparation is evidence of trans acceleration, suggesting that the movement of dopamine across the membrane is carrier mediated as opposed to being mediated via channels or pores and that the rate-limiting step in inwardly directed transport is the reorientation of the unloaded transporter from the inwardly to the outwardly facing forms.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Kinetic evaluation of the commonality between the site(s) of action of cocaine and some other structurally similar and dissimilar inhibitors of the striatal transporter for dopamine.

The inhibition by cocaine of the apparent initial rate of the transport of striatal dopamine was compared with inhibitions produced by cocaethylene, benztropine, GBR-12909, mazindol, and nomifensine. Rotating disk electrode voltammetry was used to measure the kinetically resolved, inwardly directed transport of dopamine in striatal suspensions. Evidence is presented that the primary site of action of cocaine may be at the external face of the transporter. Experiments to determine whether or not the other inhibitors bind to the same site as cocaine were conducted by comparing the inhibitions observed for each of the inhibitors alone with that observed when paired with cocaine. The resulting changes in the velocity of the transport of dopamine induced by the inhibitors were then fit to one of the previously developed models of inhibition by pairs of inhibitors affecting the kinetics of actively transporting systems: a single-site model, a two-site model in which the two binding sites for the inhibitors interact, and a two-site model in which the two binding sites for the two inhibitors act independently. Cocaine inhibited the transport of dopamine competitively with its structural analogues, cocaethylene and benztropine. The structurally dissimilar inhibitor, GBR-12909, was found also to be competitive with cocaine. In contrast, mazindol and nomifensine were found to bind to separate interactive sites when individually paired with cocaine. These results suggest that mazindol and nomifensine may interact with the kinetically active transporter for dopamine in a manner different from that of cocaine. Mazindol was tested and found to inhibit competitively the inward transport of dopamine into striatal suspensions. In contrast, our previous published findings show cocaine to be an uncompetitive inhibitor of the transport of striatal dopamine. These results suggest that cocaine inhibits inward transport of dopamine by reducing the intramembrane turnover of the transporter, whereas mazindol alters the kinetics of the recognition of dopamine by the transporter. Finally, the potential effects of these binding modes of inhibitors on synaptic chemical communication in dopaminergic systems were analyzed. The results of these analyses suggest that different effects on the extracellular concentrations of dopamine can result from the different patterns of inhibition, suggesting that different modulatory influences on pre- and postsynaptic receptor occupation can result from inhibition of the transport of dopamine.

Animals↗
PubMed1994

Individual locomotor response to novelty predicts selective alterations in D1 and D2 receptors and mRNAs.

Rats that have a greater locomotor response to novelty (high responders, HR) have differences in measures of presynaptic dopamine transmission compared to low responders (LR) to a novel environment, including altered dopamine release and behavioral response to indirect dopamine agonists. This study examined the role of three dopamine terminal fields, the nucleus accumbens, striatum, and medial prefrontal cortex, in differences between HR and LR. In the first experiment, dopamine was infused directly into the nucleus accumbens (0, 3, 10, and 30 micrograms/side) or the striatum (0, 10, 30, and 100 micrograms/side). HR showed a greater behavioral response to both the 3 and 30 micrograms/side doses infused into the nucleus accumbens compared to LR. No differences between HR and LR were revealed by dopamine infusion into the striatum. In the second experiment, radioligand binding assays were performed to determine if differences exist between high and low responder rats in the Bmax and/or KD of radiolabeled antagonist ligands for the dopamine D1 and/or D2 receptors. There were fewer D2 binding sites in the nucleus accumbens and fewer sites in the striatum in HR compared to LR. High responders showed a greater Bmax for D1 binding sites in the nucleus accumbens than LR. No differences in number of binding sites for D1 receptors were observed between HR and LR in the striatum. No differences between HR and LR in D2 or D1 receptor binding were observed in the medial prefrontal cortex. There were no differences in KD for any of the dopamine receptors in the regions examined.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

D2 receptors may modulate the function of the striatal transporter for dopamine: kinetic evidence from studies in vitro and in vivo.

Recently it was hypothesized by others that the D2 dopamine receptor can regulate the uptake of dopamine. However, the evidence in support of this hypothesis, although compelling, was not based on observations related to direct measures of the kinetic activity of the transporter itself. Here kinetic evidence in support of this hypothesis is shown. The apparent time-resolved initial velocity of the transport of 1.0 microM dopamine into striatal suspensions, measured using rotating disk electrode voltammetry, was found to increase in the presence of the D2 receptor agonist, quinpirole, at 100 nM. This effect was reversed by sulpiride. In separate studies it was shown that acute and chronic treatments with haloperidol at 0.5 mg/kg, i.p., reduced the reuptake transport of dopamine in vivo following intrastriatal stimulation of its release by K+. Thus, it appears that D2 receptors may influence the functioning of the striatal transporter for dopamine. These results are consistent with a model in which presynaptically released dopamine may feed back onto the function of its transporter to increase the velocity of the clearance of synaptic dopamine following an action potential, suggesting the existence of a mechanism, in addition to release and synthesis modulation, for fine-tuning dopaminergic chemical signaling.

Animals↗
PubMed1992

A multisubstrate mechanism of striatal dopamine uptake and its inhibition by cocaine.

A study of Na+ and Cl- as co-substrates in dopamine uptake into striatal suspensions and inhibition of dopamine uptake by cocaine was made by monitoring the initial velocity of the uptake of exogenously added non-radioactively labeled dopamine using a rotating disk electroanalytical technique with 50 msec resolution. Dopamine, in the concentration range of 0.025 to 4.00 microM, was found to be taken up rapidly into the tissue phase of striatal suspensions following the apparent zero order rate law for the first 25 sec. The observed, dopamine concentration-dependent, initial velocity data were first analyzed graphically using the Eadie-Hofstee transformation of the Michaelis-Menten kinetic equation and, subsequently, using all of the velocity data and the results of the graphical analyses, by non-linear curve fitting. Dopamine uptake was found to be first order in dopamine with a Vmax of 582 pmol/sec/g wet weight and a Km of 1.2 microM. The results of experiments in which choline and isethionate were substituted for Na+ and Cl-, respectively, suggested that the uptake process is second order in Na+ and first order in Cl-. Multisubstrate analyses of the initial velocities of uptake over the concentration range of 0.025 to 1.5 microM dopamine suggested that the mechanism of binding of dopamine to the uptake carrier is a partially random, sequential mechanism where dopamine or Na+ binds first with the uptake carrier and Cl- binds last. Cocaine was found to uncompetitively inhibit dopamine uptake and competitively inhibit both Na+ and Cl- binding (apparent Km values: 131 and 51 mM, respectively), suggesting that the mechanism of cocaine inhibition may be to bind to the dopamine occupied uptake carrier complex at the Na+ binding site.

Animals↗