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Analysis of Homogeneous Populations of Anterior Pituitary Folliculostellate Cells by Laser Capture Microdissection and Reverse Transcription-Polymerase Chain Reaction¹

Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota 55905

Address all correspondence and requests for reprints to: Dr. R. V. Lloyd, Department of Laboratory Medicine and Pathology, 200 1st Street SW, Rochester, Minnesota 55905. E-mail: lloyd.ricardo{at}mayo.edu

Pituitary folliculostellate (FS) cells are usually located between the secretory cells in the anterior pituitary, and they produce many peptides that exert a paracrine effect on hormone-producing pituitary cells. Previous approaches have been unsuccessful in obtaining homogeneous populations of FS cells. We used a combination of immunostaining with S100 protein followed by laser capture microdissection (Immuno-LCM) to obtain purified populations of rat FS cells. These cells were analyzed along with a mouse FS cell line (TtT/GF) by RT-PCR for gene expression.

RT-PCR analyses showed that both FS cell populations expressed the mRNAs for glial fibrillary acidic protein, S100 protein, transforming growth factor-ß1 (TGFß1), TGFß receptor, interleukin-6, leptin, leptin receptor, pituitary adenylate cyclase-activating polypeptide (PACAP), and PACAP receptors. Both FS cell populations were negative for PRL, GH, and POMC, supporting the homogeneity of the rat FS cell population. TGFß1, but not PACAP-38, treatment stimulated cell proliferation in both FS cell populations. TGFß1 increased leptin, but not interleukin-6, mRNA expression in rat FS cells. However, TGFß1 inhibited leptin RNA expression in the TtT/GF cell line, as shown by RT-PCR and Northern blot analysis.

These results indicate that 1) homogeneous populations of FS cells can be prepared by Immuno-LCM; 2) TGFß1 stimulates the proliferation of normal rat FS cells and the TtT/GF cell line; and 3) the effects of TGFß1 to stimulate leptin mRNA expression in rat FS cells but inhibit leptin mRNA expression in TtT/GF cells probably reflect alterations in signal transduction in the TtT/GF cell line.