Spectral analyses of extrinsic fluorescence of the nerve membrane labeled with aminonaphthalene derivatives

Tasaki, Ichiji; Carbone, Emilio; Sisco, Kenneth L.; Singer, Irwin

doi:10.1016/0005-2736(73)90146-6

Cited by 37 publications

(19 citation statements)

References 18 publications

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“…The dotted line in the figure fits the experimental data obtained by subtraction of the emission spectra of 2,6-TNS in 100% ethanol, and 80% ethanol plus 20% D20 (see ref. [16]). …”

Section: Demonstration Of Involvement Of Water In Nerve Excitationmentioning

confidence: 93%

“…Emission spectra of internally labeled axons bathed in artificial sea water at 5 and 19 ~ were obtained by the use of the spectrofluorometer previously described [16]. Lowering the temperature of the external bath was found to bring about an increase in the resting fluorescence intensity associated with a blue shift in the wavelength of emission maximum and a decrease in the haft-peak bandwidth.…”

Section: Temperature Effects On the Fluorescence Of 26-tnsmentioning

confidence: 99%

“…To carry out nanosecond lifetime measurements at different temperatures, a cooling device was constructed inside the sample housing Detection of changes in fluorescence intensity and spectral analyses of fluorescence signals (i.e., intensity changes associated with nerve excitation) were performed by using a spectrofluorometer specifically designed to studies of the nerve membrane [16]. The optical setup, the nerve chamber, and the electrical recording device employed were similar to those used in previous studies and have already been described elsewhere in detail [16,19]. The same spectrofluorometer was used to determine the emission spectra of internally labeled axons at different temperatures.…”

Section: Optical and Electronic Apparatusmentioning

confidence: 99%

“…This study is intended to be an extension of observations previously reported [16] and was carried out to elucidate the possible involvement of water in the process of nerve excitation. The data obtained are interpreted on the basis of the results of in vitro investigations demonstrating the effects of DzO and H20 on the fluorescence of 2,6-TNS in organic solvents.…”

mentioning

confidence: 97%

“…Among all the fluorescent probe-molecules so far employed, aminonaphthalene derivatives have yielded the most interesting results. Recently, particular attention has been given to the physico-chemical properties of 2-p-toluidinyl-6-naphthalene sulfonate (2,6-TNS) and emphasis placed on the information it can furnish about the macromolecular organization of the axonal membrane [ 15,16]. However, up…”

mentioning

confidence: 99%

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Physico-chemical properties of 2,6-TNS binding sites in squid giant axons: Involvement of water molecules in the excitation process

1974

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Summary. The lifetime of the excited singlet state of 2,6-TNS (2-p-toluidinyl-6-naphthalene sulfonate) in the squid giant axon was measured by the use of the single photon counting technique. The accessibility of water molecules to the bound 2,6-TNS molecules by the use of heavy water indicated that these binding sites are only partially exposed to surrounding water molecules. Studies of the effect of temperature changes on the fluorescence of 2,6-TNS in axons suggested that relaxation of polar solvent molecules and/or neighboring polar side groups (during the lifetime of the excited dye molecule) plays a crucial role in the fluorescence emission from the bound 2,6-TNS. Time-resolved emission spectra suggested that the viscosity of the microenvironment of the axoplasmic bound dye molecules is high. The effect of replacing HzO with DzO in the medium on the fluorescence signals (intensity changes) from internally labeled squid giant axons during nerve excitation were investigated. The fluorescence signals (31) in the D20 were found to be smaller in size and longer in duration than those in HzO. The spectrum of A1 in DzO was similar to that in HzO; that is to say, blue-shifted and narrow as compared with the fluorescence spectrum from the axon at rest.The method of determining extrinsic fluorescence has been applied in the past as a means of studying structural changes of membrane macromolecules during the process of nerve excitation [17,19,20]. Among all the fluorescent probe-molecules so far employed, aminonaphthalene derivatives have yielded the most interesting results. Recently, particular attention has been given to the physico-chemical properties of 2-p-toluidinyl-6-naphthalene sulfonate (2,6-TNS) and emphasis placed on the information it can furnish about the macromolecular organization of the axonal membrane [ 15,16]. However, up

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“…The dotted line in the figure fits the experimental data obtained by subtraction of the emission spectra of 2,6-TNS in 100% ethanol, and 80% ethanol plus 20% D20 (see ref. [16]). …”

Section: Demonstration Of Involvement Of Water In Nerve Excitationmentioning

confidence: 93%

Section: Temperature Effects On the Fluorescence Of 26-tnsmentioning

confidence: 99%

Section: Optical and Electronic Apparatusmentioning

confidence: 99%

mentioning

confidence: 97%

mentioning

confidence: 99%

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Physico-chemical properties of 2,6-TNS binding sites in squid giant axons: Involvement of water molecules in the excitation process

1974

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Organische Fluoreszenzreagentien für die Untersuchung von Enzymen und Proteinen

Kanaoka

1977

Angewandte Chemie

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Organic Fluorescence Reagents in the Study of Enzymes and Proteins

Kanaoka

1977

Angew. Chem. Int. Ed. Engl.

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Studies designed to elucidate life processes require close cooperation between various scientific disciplines. A pertinent example is seen in the application of fluorescence spectroscopy to biological studies, where biological sciences, physical and organic chemistry, and technical innovations complement one another. This report reviews the application of fluorescence probes which bind covalently to certain sites of proteins. The major organic fluorescence reagents used in this field are tabulated.

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Spectral analyses of extrinsic fluorescence of the nerve membrane labeled with aminonaphthalene derivatives

Cited by 37 publications

References 18 publications

Physico-chemical properties of 2,6-TNS binding sites in squid giant axons: Involvement of water molecules in the excitation process

Physico-chemical properties of 2,6-TNS binding sites in squid giant axons: Involvement of water molecules in the excitation process

Organische Fluoreszenzreagentien für die Untersuchung von Enzymen und Proteinen

Organic Fluorescence Reagents in the Study of Enzymes and Proteins

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