Neuroanatomical methods for the quantitative evaluation of coexistence of transmitters in nerve cells. Analysis of the ACTH- and beta-endorphin immunoreactive nerve cell bodies of the mediobasal hypothalamus of the rat.

A new statistical approach has been introduced to study in a quantitative way the coexistence of two neuromodulators in nerve cell bodies. The method has been exemplified on the ACTH-like and beta-endorphin-like immunoreactive nerve cell bodies of the mediobasal hypothalamus demonstrated by means of indirect immunofluorescence methodology. The method is based on the analysis of 3 adjacent sections which, in a random way, are stained with antiserum against neuromodulator 1, against neuromodulator 2 and with antisera against both neuromodulator 1 and neuromodulator 2. It could be shown that some neurons of the mediobasal hypothalamus do not contain at a detectable level both ACTH- and beta-endorphin-like immunoreactivity. The present method also involves an analysis of the two nerve cell groups by means of a morphometric procedure to collect additional information on the extent of coexistence. Thus, by this approach the gravity centers of the two cell groups can be calculated. The distances between the two gravity centers of the ACTH and beta-endorphin positive cell groups were significantly different at certain levels. The present method offers unique possibilities in increasing our understanding of the functional significance of coexistence of neuromodulators in one and the same nerve cell body since it makes possible a quantitative evaluation of coexistence. The usefulness of the present method is illustrated by the findings of a possible differential synthesis of ACTH- and beta-endorphin-like material in certain cell bodies of the mediobasal hypothalamus.