Neurotransmitters monitoring

Electrochemical detection is one of the most common methods for neurotransmitter monitoring. Many neurotransmitters are... 

Denuault, in his review [8], defined the term microelectrode as an electrode with at least one dimension in the range of 0.1 to 50 pm. The small size of microelectrodes makes them possible to be used for in vivo detection, which is usually performed with very small volumes of samples, such as those for neurotransmitter monitoring in the brain. Moreover, due to its small size, at... 

Because of their strategic localization, astrocytes play a crucial role in maintaining the extracellular ionic homeostasis, provide energetic metabolites to neurons and remove excess of neurotransmitter in schedule with synaptic activity. In addition, the strategic location of astrocytes allows them to carefully monitor and control the level of synaptic activity. Indeed, number of papers during the last 15 years have shown that cultured astrocytes can respond to a variety of neurotransmitters with a variety of different patterns of intracellular calcium increases (Verkhratsky et al. 1998). Later on, studies performed in intact tissue preparations (acute brain slices) further established that the plasma membrane receptors can sense external inputs (such as the spillover of neurotransmitters during intense synaptic activity) and transduce them as intracellular calcium elevations, mostly via release of calcium from internal stores (Dani et al. 1992 Murphy et al. 1993 Porter and McCarthy... 

Figure 1.9 Comparison of the effects of an endogenously released and exogenously applied neurotransmitter on neuronal activity (identity of action). Recordings are made either of neuronal firing (extracellularly, A) or of membrane potential (intracellularly, B). The proposed transmitter is applied by iontophoresis, although in a brain slice preparation it can be added to the bathing medium. In this instance the applied neurotransmitter produces an inhibition, like that of nerve stimulation, as monitored by both recordings and both are affected similarly by the antagonist. The applied neurotransmitter thus behaves like and is probably identical to that released from the nerve...

One limitation of this method is that the specific activity of the radiolabel is progressively diluted as the radiolabelled transmitter is released from neurons and replaced by that derived from unlabelled substrate. This method also assumes that there is no compartmentalisation of the terminal stores, yet there is ample evidence that newly synthesised acetylcholine and monoamines are preferentially released. An alternative approach is to monitor the rate at which the store of neurotransmitter is depleted after inhibition of its synthesis (Fig. 4.1). However, the rate of release of some neurotransmitters (e.g. 5-HT) is partly governed by their rate of synthesis and blocking synthesis blunts release. 

Heterocyclic fluorophores based on the benzoxadiazole nucleous, namely 4-nitrobenz-2-oxa-l,3-diazole (NBD) 14 derivatives/analogs, have been widely used as derivatization reagents for analysis purposes. Examples include the amino- or thiol reactive 4-fluoro-7-nitrobenz-2-oxa-l,3-diazole (NBD-F) 15 and 4-chloro-7-nitrobenz-2-oxa-1,3-diazole (NBD-C1) 16 [45-50] and the thiol-reactive /V-((2-(iodoacetoxy)ethyl)-/V-methyl)amino-7-nitrobenz-2-oxa-1,3-dia-zole (IANBD ester) 17 [51] and 7-chlorobenz-2-oxa-l,3-diazole-4-sulfonate (SBD-C1) 18 [52], NBD-F and NBD-C1 derivatives can be excited at about 470 nm by using the relatively inexpensive and reliable argon ion lasers or newer diode pumped solid state (DPSS) lasers. NBD-F has been used as a labeling tag in various capillary electrophoresis (CE) experiments for amino acids [53-57] including the monitorization of in vivo dynamics of amino acids neurotransmitters [58]. 

O Brien KB, Esguerra M, Miller RF, Bowser MT (2004) Monitoring neurotransmitter release from isolated retinas using online microdialysis-capillary electrophoresis. Anal Chem 76 5069-5074... 

Grunewald, M. and Kanner, B. I. (1995) Conformational changes monitored on the glutamate transporter GLT-1 indicate the existence of two neurotransmitter-bound states../. Biol. Chem. 270,17017-17024. 

With the introduction of modern electronics, inexpensive computers, and new materials there is a resurgence of voltammetric techniques in various branches of science as evident in hundreds of new publications. Now, voltammetry can be performed with a nano-electrode for the detection of single molecular events [1], similar electrodes can be used to monitor the activity of neurotransmitter in a single living cell in subnanoliter volume electrochemical cell [2], measurement of fast electron transfer kinetics, trace metal analysis, etc. Voltammetric sensors are now commonplace in gas sensors (home CO sensor), biomedical sensors (blood glucose meter), and detectors for liquid chromatography. Voltammetric sensors appear to be an ideal candidate for miniaturization and mass production. This is evident in the development of lab-on-chip... 

Bowser MT, Kennedy RT. 2001. In vivo monitoring of amine neurotransmitters using microdialysis with online capillary electrophoresis. Electrophoresis 22(17) 3668-3676. 

Zhou SY, Zuo H, Stobaugh JE, Lunte CE, Lunte SM. 1995. Continuous in vivo monitoring of amino acid neurotransmitters by microdialysis sampling with on-line derivatization and capillary electrophoresis separation. Anal Chem 67(3) 594-599. 

A. J. Cunningham and J. B. Justice, Jr., Approaches to Voltammetric and Chromatographic Monitoring of Neurochemicals in Vivo, J. Chem. Ed. 1987, 64, A34. Another Nafion-coated electrode can detect 10 10 mol of the neurotransmitter nitric oxide within a single cell [T. Malinski and Z. Taha, Nitric Oxide Release from a Single Cell Measured in Situ by a Porphyrinic-Based Microsensor, Nature 1992, 358, 676]. 

K. B. O Brien, M. Esguerra, R. F. Miller, and M. T. Bowser, Monitoring Neurotransmitter Release from Isolated Retinas Using Online Microdialysis-Capillaiy Electrophoresis, Anal. Chem. 2004, 76, 5069. 

Carbon electrodes exhibit a wide range of electron transfer rates for benchmark redox systems, depending on carbon material and surface history. Two examples are shown in Figure 10.2, which compares two carbon surfaces with very different k° for Fe(CN) /4. In some cases, the variations in electrode kinetics have been particularly important to analytical applications. For example, carbon paste and carbon fiber electrodes have been used to monitor neurotransmitters in living animal brains [5,6]. The determination of catechol transmitters in the presence of relatively large amounts of interferents (e.g., ascorbate) de-... 

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