In this study we demonstrated that individual melanotropes expressed differing levels of POMC mRNA. Interspersed among the melanotropes was a small population of process-bearing cells expressing GFAP. We compared cellular resolution and feasibility for quantitation of hybrids formed by oligonucleotide probes labeled with two different markers using in situ hybridization histochemistry. In hybridizations using [35S]-labeled probes the signal could not easily be assigned to individual melanotropes, although digoxigenin-labeled probes provided good cellular resolution. Heterogeneous staining intensities of individual melanotropes for POMC mRNA were measured, and relative quantitation of changes in POMC mRNA levels following interactions with the D2 dopamine receptor was performed. We combined in situ hybridization histochemistry with immunohistochemistry to demonstrate that cells within the lobe parenchyma not expressing POMC mRNA were immunoreactive for GFAP.
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