Abstract
In situ hybridization represents a powerful technique to localize DNA or RNA of interest at the chromosomal or cellular level. In endocrine tissues composed of diverse and varied cell types, in situ hybridization has allowed the identification of specific cells responsible for the expression of genes controlling the function of the tissue. Our laboratory has routinely used this approach to understand the cellular expression of genes associated with the growth of the ovarian follicle, rupture of the follicle, and transformation of the ruptured follicle into the corpus luteum. The current study outlines the procedural details of in situ detection of mRNA in tissues and illustrates the utility of this approach in identifying the ovarian cells expressing the matrix metalloproteinases and their endogenous inhibitors, the TIMPs, in the human ovary.
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References
John HA, Birnstiel ML, Jones KW 1969 RNA−DNA hybrids at the cytological level. Nature 223 582−587
Gall JG, Pardue ML 1969 Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc Natl Acad Sci U S A 63:378−383
Warford A, Lauder I 1991 In situ hybridisation in perspective. J Clin Pathol 44:177−181
Wilcox JN 1993 Fundamental principles of in situ hybridization. J Histochem Cytochem 41: 1725−1733
Emson PC 1993 In-situ hybridization as a methodological tool for the neuroscientist. Trends Neurosci 16:9−16
Curry TE, Jr., Nothnick WB 1996 Changes in ovarian tissue inhibitor of metalloproteinase (TIMP) expression during follicular growth, ovulation, and the luteal period in the rat. Proceedings of Inhibitors of Metalloproteinases in Development and Disease. Banff: 119−126
Hagglund AC, Ny A, Leonardsson G, Ny T 1999 Regulation and localization of matrix metalloproteinases and tissue inhibitors of metalloproteinases in the mouse ovary during gonadotropin-induced ovulation. Endocrinology 140:4351−4358
Nothnick WB, Soloway P, Curry TE, Jr. 1997 Assessment of the role of tissue inhibitor of metalloproteinase-1 (TIMP-1) during the periovulatory period in female mice lacking a functional TIMP-1 gene. Biol Reprod 56:1181−1188
Simpson KS, Byers MJ, Curry TE, Jr. 2000 Spatiotemporal mRNA expression of the tissue inhibitors of metalloproteinases (TIMPs) in the ovary throughout the rat estrous cycle. Endocrinology 142:2058−2069
Liu K, Olofsson JI, Wahlberg P, Ny T 1999 Distinct expression of gelatinase A [matrix metalloproteinase (MMP)-2], collagenase-3 (MMP-13), membrane type MMP 1 (MMP-14), and tissue inhibitor of MMPs Type 1 mediated by physiological signals during formation and regression of the rat corpus luteum. Endocrinology 140:5330−5338
Curry TE, Jr., Song L, Wheeler SE 2001 Cellular localization of gelatinases and tissue inhibitors of metalloproteinases during follicular growth, ovulation and early luteal formation in the rat. Biol Reprod 65:855−865
Acknowledgements
We are indebted to Ms. Sarah Wheeler-Price and Ms. Lauren Thomas for their assistance with the in situ hybridization. This work was supported by grants to TEC (NIH HDO57446 and NCRR P20 RR15592).
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Rosewell, K.L., Curry, T.E. (2009). Detection of Ovarian Matrix Metalloproteinase mRNAs by In Situ Hybridization. In: Park-Sarge, OK., Curry, T. (eds) Molecular Endocrinology. Methods in Molecular Biology, vol 590. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-378-7_7
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DOI: https://doi.org/10.1007/978-1-60327-378-7_7
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Publisher Name: Humana Press, Totowa, NJ
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