Biomedical subjects

D J Michael

Publications and source records attributed to D J Michael.

5 recordsLinked to original sources

PubMed2000

Subsecond adsorption and desorption of dopamine at carbon-fiber microelectrodes.

High-repetition fast-scan cyclic voltammetry and chronoamperometry were used to quantify and characterize the kinetics of dopamine and dopamine-o-quinone adsorption and desorption at carbon-fiber microelectrodes. A flow injection analysis system was used for the precise introduction and removal of a bolus of electroactive substance on a sub-second time scale to the disk-shaped surface of a microelectrode that was fabricated from a single carbon fiber (Thornel type T650 or P55). Pretreatment of the electrode surfaces consisted of soaking them in purified isopropyl alcohol for a minimum of 10 min, which resulted in S/N increasing by 200-400% for dopamine above that for those that were soaked in reagent grade solvent. Because of adsorption, high scan rates (2,000 V/s) are shown to exhibit equivalent S/N ratios as compared to slower, more traditional scan rates. In addition, the steady-state response to a concentration bolus is shown to occur more rapidly when cyclic voltammetric scans are repeated at short intervals (4 ms). The new methodologies allow for more accurate determinations of the kinetics of neurotransmitter release events (10-500 ms) in biological systems. Brain slice and in vivo experiments using T650 cylinder microelectrodes show that voltammetrically measured uptake kinetics in the caudate are faster using 2,000 V/s and 240 Hz measurements, as compared to 300 V/s and 10 Hz.

Adsorption↗

PubMed2000

Differential quantal release of histamine and 5-hydroxytryptamine from mast cells of vesicular monoamine transporter 2 knockout mice.

The recent availability of mice lacking the neuronal form of the vesicular monoamine transporter 2 (VMAT2) affords the opportunity to study its roles in storage and release. Carbon fiber microelectrodes were used to measure individual secretory events of histamine and 5-hydroxytryptamine (5-HT) from VMAT2-expressing mast cells as a model system for quantal release. VMAT2 is indispensable for monoamine storage because mast cells from homozygous (VMAT2(-/-)) mice, while undergoing granule-cell fusion, do not release monoamines. Cells from heterozygous animals (VMAT2(+/-)) secrete lower amounts of monoamine per granule than cells from wild-type controls. Investigation of corelease of histamine and 5-HT from granules in VMAT2(+/-) cells revealed 5-HT quantal size was reduced more than that of histamine. Thus, although vesicular transport is the limiting factor determining quantal size of 5-HT and histamine release, intragranular association with the heparin matrix also plays a significant role.

Animals↗

PubMed2000

Extracellular dopamine dynamics in rat caudate-putamen during experimenter-delivered and intracranial self-stimulation.

Intracranial self-stimulation is an operant behavior whereby animals are conditioned to press a lever in order to receive an electrical stimulation of their dopamine neurons. This paradigm is thought to stimulate brain reward pathways and, as such, has been used to clarify the role of dopamine in reward. Striatal extracellular dopamine concentrations were monitored during the acquisition and maintenance of self-stimulation and compared to dopamine release generated by experimenter-delivered and yoked stimulation. Fast-scan cyclic voltammetry in conjunction with carbon-fiber microelectrodes was used to monitor evoked dopamine release in the caudate-putamen during electrical stimulation of the substantia nigra/ventral tegmental area. The sub-second temporal resolution of fast-scan cyclic voltammetry coupled with the micron spatial resolution of the microelectrodes allowed for the measurement of dopamine neurotransmission in real-time. Single experimenter-delivered stimulations, identical to those used during self-stimulation, evoked dopamine release in the caudate-putamen both before and after the self-stimulation sessions. Likewise, yoked stimulations of the substantia nigra/ventral tegmental area delivered to animals untrained to perform self-stimulation resulted in an increase in extracellular dopamine levels. During training sessions, experimenter-delivered stimulations evoked dopamine release. However, as the animals began lever-pressing, extracellular dopamine levels subsequently declined. Taken together, these results suggest that dopamine functions as an alerting device, wherein increases in extracellular dopamine are obtained by unpredicted or novel rewarding stimuli, but not by those which can be anticipated.

Animals↗

PubMed1999

Improving data acquisition for fast-scan cyclic voltammetry.

Described is an improved data acquisition system for fast-scan cyclic voltammetry (FSCV). The system was designed to significantly diminish noise sources that were identified in previously recorded FSCV measurements for the detection of neurotransmitters. Minimized noise is necessary to observe the low concentrations of neurotransmitters that are physiologically important. The system was based on a high-speed, 16-bit AD/DA acquisition board that allowed high scan rates and better resolved the small faradaic currents which remained after background subtraction. Irregularities that occur when independent timing sources are used for generation of the voltage waveform and collection of the current can create large noise artifacts near the voltage limits during FSCV. These were eliminated by the use of a single acquisition board that generated the voltage waveform and collected the current. Noise from frequency drift of the power line was eliminated through the use of a phase-locked loop. To demonstrate the improved performance of the system, data were collected using carbon-fiber microelectrodes in a flow injection analysis system and in brain slices. This new data acquisition system performed significantly better than another system previously used in our laboratory without these features. The improved detection limits of the new system allowed clearly resolved current spikes featuring pre-release "feet" to be recorded adjacent to individual mast cells following chemical stimulation. When combined with false-color plots, the low-noise system facilitated identification of dopamine release in a freely moving animal.

Animals↗

PubMed1999

Electrochemical monitoring of biogenic amine neurotransmission in real time.

Three techniques, constant-potential amperometry, high-speed chronoamperometry, and fast-scan cyclic voltammetry, have been used extensively to investigate the rapid events associated with neurotransmission. These techniques vary in sensitivity, chemical resolution and temporal resolution. Amperometry provides the best temporal resolution but little chemical resolution. Fast-scan cyclic voltammetry provides both good temporal and chemical resolution, while high-speed chronoamperometry offers good temporal resolution and moderate chemical resolution. The amount of chemical information which is needed for a neurochemical measurement depends upon the sample. For single cells, secondary methods, such as HPLC and capillary electrophoresis, offer extensive chemical information about the contents of a cell. With this information, chemical information is not needed during the electrochemical measurement. Therefore, amperometry is employed to obtain the greatest temporal resolution. However, when using more complex biological samples, such as brain slices or in vivo implantation, there is a greater demand for chemical resolution provided by the electrochemical measurement. To bolster results, further confirmation is sought from anatomical, physiological and pharmaceutical evidence. Within this review, the three electrochemical techniques are outlined and compared. Examples are then provided of measurements which have been made in the three predominant biological samples which have been studied: single cells, brain slices and intact animals.

Animals↗