Abstract
A non-radioactive method of in situ hybridization was used to localize transforming growth factor-α mRNA in epithelial cells of collecting ducts and tubules in rat kidney tissue sections. The intensity and specificity of staining were assessed under a variety of tissue preparation conditions, including a direct comparison of paraffin against frozen sections. Under optimal conditions, both the signal strength and the cellular localization of the growth factor message were superior in paraffin sections. The staining method could also be used to localize the message in lung tissue, indicating that the procedure is generally applicable to other tissues. Our results indicate that the use of paraffin sections for non-radioactive in situ hybridization affords a number of advantages for the localization of specific messages in tissue sections.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alison, M.R., Nasim, M.M., Anilkumar, T.V. & Sarraf, C.E. (1993) Transforming growth factor-á immunoreactivity in a variety of epithelial tissues. Cell Prolif. 26, 449-60.
Blasband, A. J., Kathleen, T.R., Chen, X., Azizkhan, J.C. & Lee, D.C. (1990) Characterization of the rat transforming growth factor-α gene and identification of promoter sequences. Mol. Cell Biol. 10, 2111-21.
Clavel, C., Binninger, I., Boutterin, M.-C., Polette, M. & Birembaut, P. (1991) Comparison of four nonradioactive and 35S-based methods for the detection of human papillomavirus DNA by in situ hybridization. J. Virol. Methods 33, 253-66.
Fermin C. & Martin D. (1995) Expression of S100β in sensory and secretory cells of the vertebrate inner ear. Cell Mol Biol. 41, 213-25.
Le Guellec, P., Dumas, S., Volle, G.E., Pidoux, E., Moukhtar, M.S. & Treilhou-Lahille, F. (1993) An efFIcient method to detect calcitonin mRNA in normal and neoplastic rat C-cells (medullary thyroid carcinoma) by in situ hybridization using a digoxigeninlabeled synthetic oligodeoxyribonucleotide probe. J. Histochem. Cytochem. 41, 389-95.
Liu, J.-Y., Morris, G.F., Lei, W.-H., Corti, M. & Brody, A.R. (1996) Up-regulated expression of transforming growth factor-β in the bronchiolar-alveolar duct regions of asbestos-exposed rats. Am. J. Pathol. 149, 205-17.
Martinez-Montero, J.C., Herrington, C. S., Stickland, J., Sawyer, H., Evans, M., Flannery, D.M.J. & Mcgee, J.O.D. (1991) Model system for optimising mRNA non-isotopic in situ hybridisation: riboprobe detection of lysozyme mRNA in archival gut biopsy specimens. J. Clin. Pathol. 44, 835-9.
Panoskaltsis-Mortari, A. & Bucy, R.P. (1995) In situ hybridization with digoxigenin-labeled RNA probes: facts and artifacts. BioTechniques 18, 300-7.
Singer, R.H., Lawrence, J.B. & Villnave, C. (1986) Optimization of in situ hybridization using isotopic and non-isotopic detection methods. BioTechniques 4, 230-50.
Walker, C., Everitt, J., Freed, J. J., Knudson, Jr, A.G. & Whiteley, L.O. (1991) Altered expression of transforming growth factor-β in hereditary rat renal cell carcinoma. Cancer Res. 51, 2973-8.
Wilcox, J.N. (1993) Fundamental principles of in situ hybridization. J. Histochem. Cytochem. 41, 1725-33.
Rights and permissions
About this article
Cite this article
Hoyle, G.W. Investigation of Tissue Preparation Conditions for Non-radioactive In Situ Hybridization: Localization of Transforming Growth Factor-α Message in Rat Kidney. Histochem J 30, 793–798 (1998). https://doi.org/10.1023/A:1003425122408
Issue Date:
DOI: https://doi.org/10.1023/A:1003425122408