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4,5-diaminofluoroscein imaging of nitric oxide synthesis in crayfish terminal ganglia

Schuppe, Hansjürgen, Cuttle, Matthew, Chad, John.E. and Newland, Philip.L. (2002) 4,5-diaminofluoroscein imaging of nitric oxide synthesis in crayfish terminal ganglia Journal of Neurobiology, 53, (3), pp. 361-369. (doi:10.1002/neu.10117).

Record type: Article


We have analyzed the synthesis of nitric oxide in the terminal abdominal ganglion of the crayfish using the fluorescent probe 4,5-Diaminofluoroscein diacetate, DAF-2 DA. Following DAF-2 loading, ganglia showed cell-specific patterns of fluorescence in which the occurrence of strongly fluorescent cell bodies was highest in specific anterior, central, and posterior regions. We found that preincubation with the nitric oxide synthase (NOS) inhibitor L-NAME prevented much of the initial development of DAF-2 fluorescence, whereas the inactive isomer D-NAME had no effect. Washout of preincubated L-NAME caused increased cell-specific fluorescence due to endogenous NOS activity. Application of the NOS substrate L-arginine also resulted in an increase of DAF-2 fluorescence in a cell-specific manner. We bath applied the NO donor SNAP to increase exogenous NO levels which resulted in DAF-2 fluorescence increases in most cells. We therefore presume that the cell-specific pattern of DAF-2 fluorescence indicates the distribution of neurones actively synthesizing NO. The similarity between the DAF-2 staining pattern and previously published studies of NOS activity are discussed.

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Published date: 15 November 2002
Keywords: fluorescein analysis: ganglia, sensory chemistry: nitric oxide analysis: nitric oxide biosynthesis


Local EPrints ID: 56368
ISSN: 0022-3034
PURE UUID: 9733d63a-6a92-4496-8245-fcb1496de260
ORCID for John.E. Chad: ORCID iD

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Date deposited: 08 Aug 2008
Last modified: 17 Jul 2017 14:31

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Author: Hansjürgen Schuppe
Author: Matthew Cuttle
Author: John.E. Chad ORCID iD

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