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Science 19 April 1985:
Vol. 228. no. 4697, pp. 332 - 335
DOI: 10.1126/science.2858916
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Articles

Science, Vol 228, Issue 4697, 332-335
Copyright © 1985 by American Association for the Advancement of Science

articles

Silver-intensified gold and peroxidase as dual ultrastructural immunolabels for pre- and postsynaptic neurotransmitters

AN van den Pol

An ultrastructural immunostaining method that uses silver-intensified gold was combined with another procedure that uses biotin peroxidase conjugates to allow simultaneous identification of two neurotransmitter-related antigens in the central nervous system. Tyrosine hydroxylase-immunoreactive neurons labeled with silver-intensified gold could be differentiated at both light and electron microscopic levels from glutamate decarboxylase-immunoreactive neurons labeled with peroxidase. Cross reactivity of the second group of immunoreagents with the first group was reduced by the heavy metal silver shell formed around the colloidal gold immunoglobulin complex. With this dual pre-embedding method, peroxidase-stained axons containing the inhibitory neurotransmitter gamma-aminobutyric acid were found to synapse directly on silver-stained dopamine neurons in the rat dorsomedial hypothalamus. This approach can be used in combination with a post-embedding immunocytochemical colloidal gold procedure, allowing ultrastructural identification of three neurotransmitter-related antigens in the same tissue section.


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