Single cell glutamate analysis in Aplysia sensory neurons

Glutamate is the major excitatory neurotransmitter in the central nervous system. However, techniques and assays available for the determination and detection of glutamate are limited. Here we have applied an effective glutamate assay toward the high-throughput analysis of single neurons. Initial ph...

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Bibliographic Details
Published in:Journal of neuroscience methods 2005-05, Vol.144 (1), p.73-77
Main Authors: Drake, Timothy J., Jezzini, Sami, Lovell, Peter, Moroz, Leonid L., Tan, Weihong
Format: Article
Language:English
Subjects:
Animals
Aplysia
Aplysia - cytology
Aplysia - metabolism
Aspartic Acid - metabolism
Ganglia, Invertebrate - cytology
Glutamate
Glutamate assay
Glutamic Acid - analysis
Immunologic Techniques
Mechanosensory
Mechanotransduction, Cellular - physiology
Neurons, Afferent - metabolism
Oxazines - metabolism
Single cell analysis
Tissue Distribution
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Summary:Glutamate is the major excitatory neurotransmitter in the central nervous system. However, techniques and assays available for the determination and detection of glutamate are limited. Here we have applied an effective glutamate assay toward the high-throughput analysis of single neurons. Initial physiological studies and recent immunohistochemical data strongly suggested that mechanosensory neurons could use l-glutamate as a co-transmitter (in addition to sensorin) in the sensory–motor synapse of Aplysia californica. We have evaluated the levels of glutamate in these cells and compared them to other non-sensory Aplysia neurons. Since this is the first report of this assay in single cellular analysis, a series of chemical and cellular controls were also done. Based on our results, we were able to determine the concentration levels inside single Aplysia sensory neurons to be 29 mM, with significant heterogeneity between individual cells. In comparison to the pleural mechanosensory neurons, non-sensory abdominal neurons contained approximately 3 mM glutamate. These elevated levels in the sensory neurons confirm the earlier findings [Dale N, Kandel ER. l-Glutamate may be the fast excitatory transmitter of Aplysia sensory neurons. Proc Natl Acad Sci USA 1993;90:7163–7.], suggesting that glutamate plays a role as neurotransmitter in these cells.
ISSN:0165-0270
1872-678X
DOI:10.1016/j.jneumeth.2004.10.011