Skip to main content
Log in

Semi-quantitative estimation of changes in noradrenaline content and intraneuronal distribution in the rat vas deferens by fluorescence histochemistry

  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

A semi-quantitative histochemical assay for noradrenaline was developed, based on the assumption that the rate of reaction of noradrenaline with paraformaldehyde depends on transmitter concentration. Changes in organ noradrenaline content caused by drugs or cold-stress were associated with similar changes in fluorescence intensity of organ samples taken for microscopy. Differences in the fluorescence intensity of experimental and control tissues were also found when there was no change in total noradrenaline content, suggesting that fluorescence intensity is not a simple function of whole organ noradrenaline content. Changes in the relative fluorescence of experimental tissues with different paraformaldehyde exposures suggested that the intraneuronal distribution of noradrenaline may affect the rate of development of fluorescence. Analysis of the time course of the fluorescence reaction showed that this was best described by the sum of two first-order exponential components of different half-life. Further results suggested that the first, fast component represents vesicle-bound noradrenaline, while the slow component represents extragranular transmitter.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Agnati, L.F., Fuxe, K.: Quantitative comparisons of amine fluorescence in cortical noradrenaline terminals using smear preparations. J. Histochem. Cytochem. 22, 1122–1127 (1974)

    Google Scholar 

  • Bjorklund, A., Ehinger, B., Flack, B.: Analysis of fluorescence excitation peak ratios for the cellular identification of noradrenaline, dopamine or their mixtures. J. Histochem. Cytochem. 20, 56–64 (1972)

    Google Scholar 

  • Bjorklund, A., Falck, B.: Cytofluorometry of biogenic monoamines in the Falck-Hillarp method. Structural identification by spectral analysis. In: Fluorescence techniques in cell biology, pp. 171–181. Berlin-Heidelberg-New York: Springer 1973

    Google Scholar 

  • Blakeley, A.G.H., Dearnaley, D.P., Harrison, V.: The noradrenaline content of the vas deferens of the guinea pig. Proc. roy. Soc. B 174, 491–502 (1970)

    Google Scholar 

  • Corrodi, H., Jonsson, G.: The formaldehyde fluorescence method for the histochemical demonstration of biogenic monoamines. A review on the methodology. J. Histochem. Cytochem. 15, 65–78 (1967)

    Google Scholar 

  • Falck, B.: Observations on the possibilities of the cellular localization of monoamines by a fluorescence method. Acta physiol. scand. 56, S197 (1962)

    Google Scholar 

  • Falck, B., Hillarp, N.A., Thieme, G., Torp, A.: Fluorescence of catecholamines and related compounds condensed with formaldehyde. J. Histochem. Cytochem. 10, 348–354 (1962)

    Google Scholar 

  • Fillenz, M., West, D.P.: Changes in vesicular dopamine-β-hydroxyalase resulting from transmitter release. J. Neurochem. 23, 411–416 (1974)

    Google Scholar 

  • Gillespie, J.S., Hamilton, D.N.H.: Binding of noradrenaline to smooth muscle cells in the spleen. Nature (Lond.) 212, 524–525 (1966)

    Google Scholar 

  • Iversen, L.L.: The uptake and storage of noradrenaline in sympathetic nerves, pp. 36–37. Cambridge: Cambridge University Press 1967

    Google Scholar 

  • Jonsson, G.: Microfluorimetric studies on the formaldehyde-induced fluorescence of noradrenaline in adrenergic nerves of rat iris. J. Histochem. Cytochem. 17, 714–723 (1969)

    Google Scholar 

  • Lidbrink, P., Jonsson, G.: Semiquantitative estimation of formaldehyde-induced fluorescence of noradrenaline in central noradrenaline nerve terminals. J. Histochem. Cytochem. 19, 747–757 (1971)

    Google Scholar 

  • Lindvall, O., Bjorklund, A., Svensson, L.A.: Fluorophore formation from catecholamines and related compounds in the glyoxylic acid fluorescence histochemical method. Histochemistry 39, 197–227 (1974)

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Malmfors, T.: Fluorescent histochemical studies on the uptake, storage and release of catecholamines. Circulat. Res. 21, S3, 25–42 (1967)

    Google Scholar 

  • Norberg, K. A.: Drug-induced changes in monoamine levels in the sympathetic adrenergic ganglion cells and terminals. A histochemical study. Acta physiol. scand. 65, 221–234 (1965)

    Google Scholar 

  • Olson, L., Hamberger, B., Jonsson, G., Malmfors, T.: Combined fluorescence histochemistry and 3H-noradrenaline measurements of adrenergic nerves. Histochemie 15, 38–45 (1968)

    Google Scholar 

  • Van Orden, L. S., Schaefer, J. M., Burke, J. P., Lodoen, F. V.: Differentiation of norepinephri storage compartments in peripheral adrenergic nerves. J. Pharm. exp. Ther. 174, 357–368 (1970)

    Google Scholar 

  • Ritzen, M.: Quantitative fluorescence microspectrophotometry of catecholamine-formaldehyde products. Exp. Cell Res. 44, 505–520 (1966)

    Google Scholar 

  • Ritzen, M.: Microfluorimetric quantitation of biogenic monamines. In: Fluorescent techniques in cell biology, p. 183–189. Berlin-Heidelberg-New York Springer 1973

    Google Scholar 

  • Stanford, C.: The relation of organ noradrenaline concentration and the fluorescence histochemical reaction. J. Physiol. (Lond.) 245, 108–109 (1975)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stanford, C. Semi-quantitative estimation of changes in noradrenaline content and intraneuronal distribution in the rat vas deferens by fluorescence histochemistry. Histochemistry 50, 129–145 (1976). https://doi.org/10.1007/BF00495824

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00495824

Keywords

Navigation