Summary
Chicken embryo sympathetic ganglion fragments, grown in culture for three days, were incubated for short periods in appropriate experimental and control media to provide information on axonal uptake of exogenous noradrenaline in the presence and absence of the uptake-blocking drugs, protriptyline and phenoxybenzamine. The cultures were dried, gassed in formaldehyde vapour and examined by fluorescence microscopy. Spot readings were taken on axon-bundles using a microspectrophotometer. Readings were corrected for the amount of tissue present in each field, using graticule and interferometric methods. Brightness was uniform along individual axon-bundles, but varied from bundle to bundle within the same culture. Despite this range in brightness the readings fell into a normal distribution about the mean. Dose-dependent increases in brightness were obtained following incubation with noradrenaline. A clear indication of the effect of the blocking drugs was observed but the wide range in brightness prevented reliable quantitative data on these effects from being obtained. It was concluded that the wide range of brightness within each culture reflected a lack of equivalence in the intrinsic noradrenaline-uptake properties of the axon-bundles developing in such cultures.
Similar content being viewed by others
References
Birks, R.I.: Effects of stimulation on synaptic vesicles in sympathetic ganglia, as shown by fixation in the presence of Mg++. J. Physiol. (Lond.) 216, 26–28P (1971)
Bunge, R.P., Rees, R., Wood, P., Burton, H., Ko, C.-P.: Anatomical and physiological observations on synapses formed on isolated autonomic neurons in tissue culture. Brain Res. 66, 401–412 (1974)
Chamley, J.H., Goller, I., Burnstock, G.: Selective growth of sympathetic nerve fibres to explants of normally densely innervated autonomic effector organs in tissue culture. Develop. Biol. 31, 362–379 (1973)
Chamley, J.H., Mark, G.E., Campbell, G.R., Burnstock, G.: Sympathetic ganglia in culture. Z. Zellforsch. 135, 287–314 (1972)
England, J.H., Goldstein, H.N.: The uptake and localization of catecholamines in chick embryo sympathetic neurons in tissue culture. J. Cell Sci. 4, 677–691 (1969)
Fuxe, K., Hökfelt, T., Jonsson, G., Ungerstedt, U.: Fluorescence microscopy in neuroanatomy. In: Contemporary research methods in neuroanatomy (W.J.H. Nauta and S.O.E. Ebbesson, eds.), pp. 275–314. Berlin-Heidelberg-New York: Springer 1970
Hamburger, V., Hamilton, H.L.: A series of normal stages in the development of the chick embryo. J. Morph. 88, 49–92 (1951)
Lever, J.D., Presley, R.: Studies on the sympathetic neurone in vitro. Progr. Brain Res. 34, 499–512 (1972)
Lidbrink, P., Johnsson, G.: Semiquantitative estimation of formaldehyde-induced fluorescence of noradrenaline in central noradrenaline nerve terminals. J. Histochem. Cytochem. 19, 747–757 (1971)
Mottram, D.R., Presley, R., Lever, J.D., Ivens, C.: Some characteristics of chick embryo sympathetic ganglion fragments in a simple culture system with and without nerve growth factor. Z. Anat. Entwickl.-Gesch. 138, 127–133 (1972)
Mottram, D.R., Presley, R., Lever, J.D., Ivens, C.: Sympathetic nerve cultures as a tool for the pharmacological study of sympathetic terminals. Europ. J. Pharmacol. 22, 175–180 (1973)
Teichberg, S., Holtzman, E.: Axonal agranular reticulum and synaptic vesicles in cultured embryonic chick sympathetic neurones. J. Cell Biol. 57, 88–108 (1973)
Author information
Authors and Affiliations
Additional information
This work was supported by a grant from the Medical Research Council. We wish to thank Mrs. G. O'Shea for valuable technical assistance, and Messrs. P.F. Hire and K. Twohigg for help with the illustrations. Phenoxybenzamine was a gift from Smith, Kline and French Ltd.
Rights and permissions
About this article
Cite this article
Presley, R., Santer, R.M., Lu, KS. et al. Measurements of noradrenaline fluorescence in axons of chick embryo sympathetic ganglia grown in three day culture. Cell Tissue Res. 178, 49–60 (1977). https://doi.org/10.1007/BF00232823
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00232823