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David Sulzer

Publications and source records attributed to David Sulzer.

31 records · Page 2Linked to original sources
PubMed2003

D3 dopamine autoreceptors do not activate G-protein-gated inwardly rectifying potassium channel currents in substantia nigra dopamine neurons.

Substantia nigra (SN) dopamine neurons express D2 and D3 dopamine autoreceptors. A physiological role for the D3 receptor has not been identified, but an activation of G-protein-gated inwardly rectifying potassium (GIRK; also known as Kir3) channels is strongly implicated because D3 receptors activate channels composed of GIRK2 subunits in cell lines. We confirmed that acutely dissociated SN dopamine neurons indeed contain D3 and GIRK2 subunit mRNA using single-cell RT-PCR. We then tested whether D3 receptors activate GIRK currents in SN dopamine neurons by comparing acutely dissociated neurons from D2-/- receptor knock-out and congenic wild-type mice. In nearly all (14 of 15) wild-type SN dopamine neurons, the D2/D3 agonist quinpirole activated GIRK currents that were blocked by cesium. Quinpirole, however, elicited no GIRK currents in any SN dopamine neuron (0 of 13) derived from D2-/- receptor knock-out mice. The absence of quinpirole response was not caused by a lack of GIRK activity, because the GABAB receptor agonist baclofen continued to elicit these currents in the mutant neurons. Thus, it appears that D3 activation of GIRK currents in SN neurons does not occur or is exceedingly rare.

Animals↗
PubMed2003

Intracellular patch electrochemistry: regulation of cytosolic catecholamines in chromaffin cells.

Alterations in the cytosolic pool directly affect neurotransmitter synthesis and release and are suggested to be key factors in various neurodegenerative disorders. Although this cytosolic pool is the most metabolically active, it is miniscule compared with the amount of vesicular transmitter and has never been quantified separately. Here, we introduce intracellular patch electrochemistry (IPE), a technique that for the first time provides direct measurements of cytosolic oxidizable molecules in single mammalian cells. In amperometric mode, IPE detects total catechols, whereas in cyclic voltammetric mode, it preferentially measures catecholamines. In cultured chromaffin cells, the total cytosolic catechol concentration was 50-500 microm, of which approximately 10% were catecholamines. Reserpine, a vesicular monoamine transporter inhibitor, had no effect on the catecholamine pool but increased total catechols by fourfold to fivefold. Combined with pargyline, a monoamine oxidase inhibitor, reserpine increased catecholamine levels in the cytosol by approximately sixfold. Amphetamine induced a transient approximately fivefold accumulation of cytosolic catecholamines and a slow increase of total catechols. In cells incubated with 3,4-dihydroxy-L-phenylalanine (L-DOPA), catecholamines increased by approximately 2.5-fold and total catechols increased by approximately fourfold. Cytosolic catecholamines returned to control levels <or=10 min after L-DOPA withdrawal, whereas total catechols remained approximately twofold elevated even after a 1.5 hr incubation in L-DOPA-free media. Our data indicate that cytosolic catecholamines are strictly maintained at a defined level, and drug-induced increases in their concentrations lead to the accumulation of other catecholamine derivatives, such as DOPAC and 3,4-dihydroxyphenylethyleneglycol. These derivatives reside in the cytosol for hours after treatment and may be an underlying cause of drug-related cytotoxicity.

Adrenergic Uptake Inhibitors↗
PubMed2003

Neuromelanin of the substantia nigra: a neuronal black hole with protective and toxic characteristics.

Neuromelanin accumulates in dopaminergic neurons during normal aging, and in Parkinson's disease, neurons with this pigment are those that selectively degenerate. Intraneuronal neuromelanin could play a protective role during its synthesis by preventing the toxic accumulation of cytosolic catechol derivatives and, in addition, by its ability to scavenge reactive metals, pesticides and other toxins to form stable adducts. However, dying neurons in Parkinson's disease that release neuromelanin might induce a vicious cycle of chronic neuroinflammation and neuronal loss.

Aging↗
PubMed2003

Dopamine transport currents are promoted from curiosity to physiology.

Midbrain dopaminergic neurons maintain low ongoing "tonic" activity interrupted by high frequency bursts associated with behavioral reward. Dopamine released during bursts feeds back onto D(2) autoreceptors that depress neuronal activity. New findings from Ingram and colleagues suggest that, by contrast, tonic activity excites these neurons by activating an uncoupled Cl- conductance that is mediated by the dopamine uptake transporter. This response extends the range of states regulated by neurotransmitter feedback, and could contribute importantly to understanding amphetamine action.

Amphetamine↗
PubMed2003

Presynaptic regulation of dopaminergic neurotransmission.

The development of electrochemical recordings with small carbon-fiber electrodes has significantly advanced the understanding of the regulation of catecholamine transmission in various brain areas. Recordings in vivo or in slice preparations monitor diffusion of catecholamine following stimulated synaptic release into the surrounding tissue. This synaptic 'overflow' is defined by the amount of release, by the activity of reuptake, and by the diffusion parameters in brain tissue. Such studies have elucidated the complex regulation of catecholamine release and uptake, and how psychostimulants and anti-psychotic drugs interfere with it. Moreover, recordings with carbon-fiber electrodes from cultured neurons have provided analysis of catecholamine release and its plasticity at the quantal level.

Animals↗
PubMed2003

Real-time decoding of dopamine concentration changes in the caudate-putamen during tonic and phasic firing.

The fundamental process that underlies volume transmission in the brain is the extracellular diffusion of neurotransmitters from release sites to distal target cells. Dopaminergic neurons display a range of activity states, from low-frequency tonic firing to bursts of high-frequency action potentials (phasic firing). However, it is not clear how this activity affects volume transmission on a subsecond time scale. To evaluate this, we developed a finite-difference model that predicts the lifetime and diffusion of dopamine in brain tissue. We first used this model to decode in vivo amperometric measurements of electrically evoked dopamine, and obtained rate constants for release and uptake as well as the extent of diffusion. Accurate predictions were made under a variety of conditions including different regions, different stimulation parameters and with uptake inhibited. Second, we used the decoded rate constants to predict how heterogeneity of dopamine release and uptake sites would affect dopamine concentration fluctuations during different activity states in the absence of an electrode. These simulations show that synchronous phasic firing can produce spatially and temporally heterogeneous concentration profiles whereas asynchronous tonic firing elicits uniform, steady-state dopamine concentrations.

Action Potentials↗
PubMed2002

The identification of vesicular glutamate transporter 3 suggests novel modes of signaling by glutamate.

Quantal release of the principal excitatory neurotransmitter glutamate requires a mechanism for its transport into secretory vesicles. Within the brain, the complementary expression of vesicular glutamate transporters (VGLUTs) 1 and 2 accounts for the release of glutamate by all known excitatory neurons. We now report the identification of VGLUT3 and its expression by many cells generally considered to release a classical transmitter with properties very different from glutamate. Remarkably, subpopulations of inhibitory neurons as well as cholinergic interneurons, monoamine neurons, and glia express VGLUT3. The dendritic expression of VGLUT3 by particular neurons also indicates the potential for retrograde synaptic signaling. The distribution and subcellular location of VGLUT3 thus suggest novel modes of signaling by glutamate.

Amino Acid Transport Systems, Acidic↗
PubMed2002

Methamphetamine-induced degeneration of dopaminergic neurons involves autophagy and upregulation of dopamine synthesis.

Methamphetamine (METH) selectively injures the neurites of dopamine (DA) neurons, generally without inducing cell death. It has been proposed that METH-induced redistribution of DA from the vesicular storage pool to the cytoplasm, where DA can oxidize to produce quinones and additional reactive oxygen species, may account for this selective neurotoxicity. To test this hypothesis, we used mice heterozygous (+/-) or homozygous (-/-) for the brain vesicular monoamine uptake transporter VMAT2, which mediates the accumulation of cytosolic DA into synaptic vesicles. In postnatal ventral midbrain neuronal cultures derived from these mice, METH-induced degeneration of DA neurites and accumulation of oxyradicals, including metabolites of oxidized DA, varied inversely with VMAT2 expression. METH administration also promoted the synthesis of DA via upregulation of tyrosine hydroxylase activity, resulting in an elevation of cytosolic DA even in the absence of vesicular sequestration. Electron microscopy and fluorescent labeling confirmed that METH promoted the formation of autophagic granules, particularly in neuronal varicosities and, ultimately, within cell bodies of dopaminergic neurons. Therefore, we propose that METH neurotoxicity results from the induction of a specific cellular pathway that is activated when DA cannot be effectively sequestered in synaptic vesicles, thereby producing oxyradical stress, autophagy, and neurite degeneration.

Animals↗
PubMed2002

Resistance of alpha -synuclein null mice to the parkinsonian neurotoxin MPTP.

Parkinson's disease (PD) is most commonly a sporadic illness, and is characterized by degeneration of substantia nigra dopamine (DA) neurons and abnormal cytoplasmic aggregates of alpha-synuclein. Rarely, PD may be caused by missense mutations in alpha-synuclein. MPTP, a neurotoxin that inhibits mitochondrial complex I, is a prototype for an environmental cause of PD because it produces a pattern of DA neurodegeneration that closely resembles the neuropathology of PD. Here we show that alpha-synuclein null mice display striking resistance to MPTP-induced degeneration of DA neurons and DA release, and this resistance appears to result from an inability of the toxin to inhibit complex I. Contrary to predictions from in vitro data, this resistance is not due to abnormalities of the DA transporter, which appears to function normally in alpha-synuclein null mice. Our results suggest that some genetic and environmental factors that increase susceptibility to PD may interact with a common molecular pathway, and represent the first demonstration that normal alpha-synuclein function may be important to DA neuron viability.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed2002

Altered dopamine release and uptake kinetics in mice lacking D2 receptors.

Dysregulation of dopamine transmission is thought to contribute to schizophrenic psychosis and drug dependence. Dopamine release is regulated by D2 dopamine autoreceptors, and D2 receptor ligands are used to treat psychosis and addiction. To elucidate the long-term effects of D2 autoreceptor activity on dopamine signaling, dopamine overflow evoked by single or paired-pulse stimulation was compared in striatal slices from D2-null mutant and wild-type mice. Quinpirole, a D2/D3 receptor agonist, had no effect on evoked dopamine release in D2 mutant mice, indicating that D2 receptors are the only release-regulating receptors at the axon terminal. Dopamine release inhibition by GABA(B) receptor activation was unchanged in D2 mutant mice, suggesting that other G-protein-coupled pathways remained normal in the absence of D2 autoreceptors. Paired-pulse stimulation revealed that autoinhibition of dopamine release was maximal 500 msec after stimulation and lasted <5 sec. In D2-null mutants, dopamine overflow in response to single stimuli was severely decreased. Experiments with the uptake inhibitor nomifensine indicated that this was caused by enhanced dopamine uptake rather than reduced release. Analysis of dopamine overflow kinetics using a simulation model suggested that the enhanced uptake was caused by an increase in the maximal velocity of uptake, V(max). These results from D2-null mutant mice support the suggestion that D2 autoreceptors and dopamine transporters interact to regulate the amplitude and timing of dopamine signals.

3,4-Dihydroxyphenylacetic Acid↗
PubMed2002

Stimulation-dependent regulation of the pH, volume and quantal size of bovine and rodent secretory vesicles.

Trapping of weak bases was utilized to evaluate stimulus-induced changes in the internal pH of the secretory vesicles of chromaffin cells and enteric neurons. The internal acidity of chromaffin vesicles was increased by the nicotinic agonist 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP; in vivo and in vitro) and by high K+ (in vitro); and in enteric nerve terminals by exposure to veratridine or a plasmalemmal [Ca2+]o receptor agonist (Gd3+). Stimulation-induced acidification of chromaffin vesicles was [Ca2+]o-dependent and blocked by agents that inhibit the vacuolar proton pump (vH+-ATPase) or flux through Cl- channels. Stimulation also increased the average volume of chromaffin vesicles and the proportion that displayed a clear halo around their dense cores (called active vesicles). Stimulation-induced increases in internal acidity and size were greatest in active vesicles. Stimulation of chromaffin cells in the presence of a plasma membrane marker revealed that membrane was internalized in endosomes but not in chromaffin vesicles. The stable expression of botulinum toxin E to prevent exocytosis did not affect the stimulation-induced acidification of the secretory vesicles of mouse neuroblastoma Neuro2A cells. Stimulation-induced acidification thus occurs independently of exocytosis. The quantal size of secreted catecholamines, measured by amperometry in cultured chromaffin cells, was found to be increased either by prior exposure to L-DOPA or stimulation by high K+, and decreased by inhibition of vH+-ATPase or flux through Cl- channels. These observations are consistent with the hypothesis that the content of releasable small molecules in secretory vesicles is increased when the driving force for their uptake is enhanced, either by increasing the transmembrane concentration or pH gradients.

Adrenal Medulla↗
PubMed2002

The absolute concentration of nigral neuromelanin, assayed by a new sensitive method, increases throughout the life and is dramatically decreased in Parkinson's disease.

The concentration of neuromelanin (NM) in substantia nigra pars compacta (SNPC) has been measured in male and female normal subjects at different ages in the range 1-97 years old and in SNPC of parkinsonian patients. A very similar age trend of NM concentration was found in both sexes. In the first year of life NM was not detectable, between 10 and 20 years the NM levels were 0.3-0.8 microg/mg of SNPC, between 20 and 50 years were 0.8-2.3 microg/mg SNPC and between 50 and 90 were 2.3-3.7 microg/mg of SNPC. In parkinsonian subjects, the NM levels were 1.2-1.5 microg/mg of SNPC, which is less than 50% with respect to the age-matched controls. These data demonstrate a continuous NM accumulation in SNPC neurons during aging, the presence of large amounts of NM in SNPC and severe depletion of NM in Parkinson's disease.

Adolescent↗