Abstract
Mammalian placentas express a large number of so-called imprinted genes. Imprinting refers to mono-allelic or biased expression based on which parent contributed the allele. Many of these imprinted loci encode factors involved in growth and cell-cycle regulation, as well as maternal behavior. In general, paternally expressed genes tend to enhance growth, whereas maternally expressed genes inhibit growth. Methods are described for developing assays to test the allelic usage of a gene. The approaches described are best utilized within a system where multiple strains are available, and it is possible to perform reciprocal crosses. Only polymerase chain reaction-based methods are examined in any detail.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Tilghman, S. (1999) The sins of the fathers and mothers: genomic imprinting in mammalian development. Cell 96, 185–193.
Haig, D. (1996) Placental hormones, genomic imprinting and maternal-fetal comunication. J. Evol. Biol. 9, 357–380.
Moore, T. and Reik, W. (1996) Genetic conflict in early development: parental imprinting in normal and abnormal growth. Rev. Reprod. 1, 73–77.
Zeng, S. M. and Yankowitz, J. (2003) X-inactivation patterns in human embryonic and extra-embryonic tissues. Placenta 24, 270–275.
Sangha, K. K., Stephenson M. D., Brown, C. J., and Robinson, W. P. (1999) Extremely skewed X-chromosome inactivation is increased in women with recurrent spontaneous abortion. Am. J. Human Genet. 65, 913–917.
Constancia, M., Hemberger, M., Hughes, J., et al. (2002) Placental-specific IGFII is a major modulator of placental and fetal growth. Nature 417, 945–948.
Wake, N., Takagi, N., and Sasaki, M. (1978) Androgenesis as a cause of hydatidiform mole. J. Natl. Cancer Inst. 60, 51–57.
Judson, H., Hayward, B. E., Sheridan, E., and Bonthron, D. T. (2002) A global disorder of imprinting in the human female germ line. Nature 416, 539–542.
Rogers, J. F. and Dawson, W. D. (1970) Foetal and placental size in a Peromyscus species cross. J. Reprod. Fertil. 21, 255–262.
Vrana, P. B., Matteson, P. G., Schmidt, J. V., et al. (2001) Genomic imprinting of a placental lactogen gene in Peromyscus. Dev. Genes Evol. 211, 523–532.
Vrana, P., Guan, X.-J., Ingram, R. S., and Tilghman, S. M. (1998) Genomic imprinting is disrupted in interspecific Peromyscus hybrids. Nature Genet. 20, 362–365.
Moglabey, Y. B., Kircheisen, R., Seoud, M., El Mogharbel, N., Van den Veyver, I., and Slim, R. (1999) Genetic mapping of a maternal locus responsible for familial hydatidiform moles. Human Mol. Genet. 8, 667–671.
Vrana, P., Fossella, J. A., Matteson, P., del Rio, T., O’Neill, M. J., and Tilghman, S. M. (2000) Genetic and epigenetic incompatibilities underlie hybrid dysgenesis in Peromyscus. Nat. Genet. 25, 120–124.
Li, Y. and Behringer, R. R. (1998) Esx1 is an X-chromosome-imprinted regulator of placental development and fetal growth. Nat. Genet. 20, 309–311.
Fohn, L. E. and Behringer, R. R. (2001) ESX1L, a novel X chromosome-linked human homeobox gene expressed in the placenta and testis. Genomics 74, 105–108.
Moore, T. and Haig, D. (1991) Genomic imprinting in mammalian development: a parental tug-of-war. Trends Genet. 7, 45–49.
Auffray, C. and Rougeon, F. (1980) Purification of mouse immunoglobulin heavychain messenger RNAs from total myeloma tumor RNA. Eur. J. Biochem. 107, 303–314.
Innis, M. A., Gelfand, D. H., Sninsky, J. J., and White, T. J. (eds.) (1990) PCR Protocols: A Guide to Methods and Applications. Academic, San Diego, CA.
Innis, M. A., Gelfand, D. H., and Sninsky, J. J. (eds.) (1999) PCR Applications: Protocols for Functional Genomics. Academic, San Diego, CA.
Singer-Sam, J., LeBon, J. M., Dai, A., and Riggs, A. D. (1992) A sensitive, quantitative assay for measurement of allele-specific transcripts differing by a single nucleotide. PCR Methods Appl. 1, 160–163.
Weber, M., Hagege, H., Lutfalla, G., et al. (2003) A real-time polymerase chain reaction assay for quantification of allele ratios and correction of amplification bias. Anal. Biochem. 320, 252–258.
Bartolomei, M. S., Zemel, S., and Tilghman, S. M. (1991) Parental imprinting of the mouse H19 gene. Nature 351, 153–155.
DeChiara, T. M., Robertson, E. J., and Efstratiadis, A. (1991) Parental imprinting of the mouse insulin-like growth factor II gene. Cell 64, 849–859.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Vrana, P.B. (2006). Assays to Determine Allelic Usage of Gene Expression in the Placenta. In: Soares, M.J., Hunt, J.S. (eds) Placenta and Trophoblast. Methods in Molecular Medicine™, vol 121. Humana Press. https://doi.org/10.1385/1-59259-983-4:437
Download citation
DOI: https://doi.org/10.1385/1-59259-983-4:437
Publisher Name: Humana Press
Print ISBN: 978-1-58829-404-3
Online ISBN: 978-1-59259-983-7
eBook Packages: Springer Protocols