Inducible Nitric Oxide Synthase in the Anterior Pituitary Gland: Induction by Interferon-γ in a Subpopulation of Folliculostellate Cells and in an Unidentifiable Population of Non-hormone-secreting Cells

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Abstract
In the context of immune-endocrine relationships, we have previously shown that interferon-γ (IFN-γ) inhibits hormone secretion in anterior pituitary (AP) cell cultures. The non-hormone-secreting folliculostellate (FS) cells were found to mediate this inhibitory action. Because in the immune system IFN-γ is a strong stimulator of nitric oxide (NO) release through the induction of NO synthase (NOS), we investigated whether the inducible form of NOS (iNOS) is present in (rat) AP cell cultures, and whether its expression is stimulated by IFN-γ. Immunocytochemistry revealed that under basal in vitro conditions only a very few AP cells contained iNOS. Treatment with IFN-γ caused a sixfold rise in the number of iNOS-positive cells and augmented the intensity of the staining. The increased number of iNOS-expressing cells was paralleled by elevated production of NO. Some of the iNOS-positive cells extended cytoplasmic processes between hormone-secreting cells, which is a characteristic of FS cells. Immunostaining of FS cell-poor and FS cell-enriched populations (obtained by gradient sedimentation) also suggested the presence of iNOS in a subpopulation of FS cells. By double immunofluorescence techniques we found that about 65% of iNOS-expressing cells were positive for S-100, a marker protein for FS cells. However, around 80% of the S-100-positive cells were not labeled for iNOS. On the other hand, the majority of the S-100-negative iNOS-containing cells could not be further identified by antisera against the classical AP hormones, suggesting the presence of iNOS in a still unidentified non-hormone-secreting cell type of the AP gland. This report is the first to demonstrate the expression of the inducible form of NOS in the AP gland. IFN-γ upregulates this expression, showing that cytokines may use the same signaling mechanisms in both the immune and the endocrine system. In addition, a putative new function of a subpopulation of FS cells in the paracrine regulation of the AP gland is suggested.

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