Tetanus toxin-enhanced GABA immunoreactivity in living neurons.
Analysis of the connectivity between different neuronal cell types is dependent on an appreciation of their dendritic and axonal arborizations. A detailed study of the dendrites and axons of GABAergic neurons has been thwarted by the lack of a suitable technique for enhancing GABA immunoreactivity. This article describes a procedure using tetanus toxin which, when applied to organotypic hippocampal cultures, considerably enhances the immunoreactivity in the dendrites and axons of the GABA- and somatostatin-containing neurons and clearly demonstrates the co-localization of GABA and somatostatin immunoreactivities in the same neuron. Tetanus toxin was applied to the culture medium on Day 14 for a 24-hr period and the cultures were fixed at the end of Day 18. Tetanus toxin-treated cultures (n = 30) or untreated cultures (n = 40) were incubated for either GABA or somatostatin immunoreactivity. Tetanus toxin-treated cultures used for co-localization studies (n = 20) were incubated for both GABA and somatostatin immunoreactivity.
Mitchell
J
J
Human Morphology, University of Southampton, Southampton, United Kingdom.
D000818:Animals; D000831:Animals, Newborn; D046508:Culture Techniques; D006624:Hippocampus / Q000737:chemistry* / Q000187:drug effects; D007150:Immunohistochemistry; D009474:Neurons / Q000737:chemistry* / Q000187:drug effects; D051381:Rats; D013004:Somatostatin / Q000032:analysis; D013744:Tetanus Toxin / Q000494:pharmacology*; D005680:gamma-Aminobutyric Acid / Q000032:analysis*
D013744:Tetanus Toxin; D013004:Somatostatin; D005680:gamma-Aminobutyric Acid