Abstract
The anterior pituitary gland consists of a heterogeneous population of various cell types. To study a single cell type with a homogeneous cell population, one can perform laser capture microdissection (LCM). Because different pituitary cells have unique immunophenotypic profiles, it is possible to perform immunohistochemical staining before LCM (immuno-LCM) for the collection of a phenotypically homogeneous cell population. These techniques were developed and applied to dissociated anterior pituitary cells and cultured pituitary cells. When combined with reverse transcriptase polymerase chain reaction, it is possible to analyze gene expression in as few as one to 10 pituitary cells. We have used the immuno-LCM technique to prepare homogeneous populations of folliculostellate cells. These cells were analyzed for expression of peptides and receptors. Anterior pituitary hormones were not expressed by these cells. These results show the utility of immuno-LCM for cellular and molecular studies of gene expression.
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References
Emmert-Buck, M. R., Bonner, R. F., Smith, P. D., Chuaqui, R. F., Zhuang, Z., Goldstein, S. R., et al. (1996) Laser capture microdissection. Science 274, 998–1001.
Bonner, R. F., Emmert-Buck, M., Cole, K., Pohida, T., Chuaqui, R., Goldstein, S., and Liotta, L. A. (1997) Laser capture microdissection: molecular analysis of tissue. Science 278, 1481,1483.
Fend, F., Emmert-Buck, M. R., Chuaqui, R., Cole, K., Lee, J., Liotta, L. A., and Raffeld, M. (1999) Immuno-LCM: laser capture microdissection of immunostained frozen sections for mRNA analysis. Am. J. Pathol. 154, 61–66.
Jin, L., Thompson, C. A., Qian, X., Kuecker, S. J., Kulig, E., and Lloyd, R. V. (1999) Analysis of anterior pituitary hormone mRNA expression in immunophenotypically characterized single cells after laser capture microdissection. Lab. Invest. 79, 511–512.
Kuecker, S., Jin, L., Kulig, E., Ondraogo, G., Roche, P., and Lloyd, R. (1999) Analysis of PRL, PRL-R, TGFβ1 and TGFβ-RII gene expression in normal and neoplastic breast tissues after laser capture microdissection. Appl. Immunohistochem. Mol. Morphol. 7, 193–200.
Goldsworthy, S. M., Stockton, P. S., Trempus, C. S., Foley, J. F., and Maronpot, R. R. (1999) Effects of fixation on RNA extraction and amplification from laser capture microdissected tissue. Mol. Carcinog. 25, 86–91.
Simone, N. L., Bonner, R. F., Gillespie, J. W., Emmert-Buck, M. R., Liotta, L. A. (1998) Laser-capture microdissection: opening the microscopic frontier to molecular analysis. Trends Genet. 14, 272–276.
Suarez-Quian, C. A., Goldstein, S. R., Pohida, T., Smith, P. D., Peterson, J. I., Wellner, E., et al. (1999) Laser capture microdissection of single cells from complex tissues. Biotechniques 26, 328–335.
Jin, L., Tsumanuma, I., Ruebel, K. H., Bayliss, J. M., and Lloyd, R. V. (2001) Analysis of homogeneous populations of anterior pituitary folliculostellate cells by laser capture microdissection and reverse transcription-polymerase chain reaction. Endocrinology 142, 1703–1709.
Cocchia, D. and Miani, N. (1980) Immunocytochemical localization of the brainspecific S-100 protein in the pituitary gland of adult rat. J. Neurocytol. 9, 771–782.
Hofler, H., Walter, G. F., and Denk, H. (1984) Immunohistochemistry of folliculostellate cells in normal human adenohypophyses and in pituitary adenomas. Acta Neuropathol. (Berl.) 65, 35–40.
Girod, C., Trouillas, J., and Dubois, M. P. (1985) Immunocytochemical localization of S-100 protein in stellate cells (folliculo-stellate cells) of the anterior lobe of the normal human pituitary. Cell Tissue Res. 241, 505–511.
Lloyd, R. V. and Mailloux, J. (1988) Analysis of S-100 protein positive folliculostellate cells in rat pituitary tissues. Am. J. Pathol. 133, 338–346.
Baes, M., Allaerts, W., and Denef, C. (1987) Evidence for functional communication between folliculo-stellate cells and hormone-secreting cells in perifused anterior pituitary cell aggregates. Endocrinology 120, 685–691.
Allaerts, W., Carmeliet, P., and Denef, C. (1990) New perspectives in the function of pituitary folliculo-stellate cells. Mol. Cell Endocrinol. 71, 73–81.
Marin, F., Stefaneanu, L., and Kovacs, K. (1991) Folliculo-stellate cells of the pituitary. Endocr. Pathol. 2, 180–192.
Ferrara, N. and Henzel, W. J. (1989) Pituitary follicular cells secrete a novel heparin-binding growth factor specific for vascular endothelial cells. Biochem Biophys. Res. Commun. 161, 851–858.
Ferrara, N., Schweigerer, L., Neufeld, G., Mitchell, R., and Gospodarowicz, D. (1987) Pituitary follicular cells produce basic fibroblast growth factor. Proc. Natl. Acad. Sci. USA 84, 5773–5777.
Vankelecom, H., Matthys, P., Van Damme, J., Heremans, H., Billiau, A., and Denef, C. (1993) Immunocytochemical evidence that S-100-positive cells of the mouse anterior pituitary contain interleukin-6 immunoreactivity. J Histochem Cytochem 41, 151–156.
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Lloyd, R.V. et al. (2005). Analysis of Pituitary Cells by Laser Capture Microdissection. In: Murray, G.I., Curran, S. (eds) Laser Capture Microdissection. Methods in Molecular Biology™, vol 293. Humana Press. https://doi.org/10.1385/1-59259-853-6:233
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DOI: https://doi.org/10.1385/1-59259-853-6:233
Publisher Name: Humana Press
Print ISBN: 978-1-58829-260-5
Online ISBN: 978-1-59259-853-3
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