Further studies on the identification of neurons containing immunoreactive alpha-melanocyte-stimulating hormone (alpha-MSH) in the rat brain.

In order to identify more accurately the organelles containing immunoreactive alpha-melanocyte-stimulating hormone (alpha-MSH) in the rat brain, attempts were made to improve the ultrastructural preservation of the neural tissue used for immunostaining of the peptide. With the post-embedding staining technique, the quality of the preservation was not adequate since only low concentrations (0.2-0.5%) of OsO4 could be used for postfixation. Higher concentrations of OsO4 completely destroyed the antigenicity of alpha-MSH. With the pre-embedding technique in which the immunostaining is performed prior to postfixation with OsO4, a good preservation could be obtained making possible the identification of organelles and classification of endings containing immunoreactive alpha-MSH. In the dendrites, the staining was rather diffuse without any clear association with organelles. In the endings, the staining was mostly restricted to dense core vesicles with some degree of diffusion in the cytoplasm. None of the positive endings were seen making synaptic contact. These results support the hypothesis that alpha-MSH could be released in sites other than the classical synaptic junction and act as a local hormone.