The heterogeneous distribution of TH, vGLUT1, and vGLUT2 within a brain region rendered a unique challenge for stereological sampling, but it emphasizes that stereological analyses remain valid even though regional variation exists in terms of intensity or distribution of the signals. Immuno-staining for tyrosine hydroxylase (TH) labeled DA terminals, and vesicular glutamate transporter 1 (vGLUT1)/vesicular glutamate transporter 2 (vGLUT2) labeled GLU terminals, which delineated regional boundaries of subdivisions in the extended amygdala due to their innate expression. Stereology was used to count the vesicular glutamate transporters and tyrosine hydroxylase-immunostained terminals in a study investigating the effect of chronic or repeated deprivation of alcohol on the dopamine (DA) and glutamate (GLU) terminals in the extended amygdala ( 4). As an alternative to purely counting neurons, one may choose to study neuronal terminals or varicosities.
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