Preparation of probes which lead to the Identification genes that are differentially expressed with retinoic acid (RA) treatment in early Xenopus laevis embryos.
Keywords:
Retinoic acid, Xenopus embryos, M-probeAbstract
Total RNA from both treated and untreated stage 14x.laevis embryos was extracted and analysed by DDRT-PCR. The RNA was coverted into CDNA by reverse transcription-PCR by using oligo (dT) primers and amplified by arbitrary 5'primers. The PCR products were compared side by side and analysed on 6% DNA sequencing gel and autoradiography. Plasmid DNA sequencing of cloned cDNAs were determined for one differential fragment (M-band). M-fragment sequence
(279bp) which was isolated from RA-treated stage 14x.laevis cDNA was tailed with poly (A) characteristic of 3'untranslated region and no significant open reading frames were present. A search of the EMBL/Gen bank databases using FASTA program appeared that the 3'end of M-cloned (86bp) has 81% homology to the xenopus laevis cyclin-dependent kinase 2 promoter (CDK2). The reverse transcription-polymerase chain reaction (RT-PCR) analysis appears to support the idea that the quantity of M-mRNAs is increased in the presence of retinoic acid. The 279bp of M-clone was stored at -20°C to use it as a probe for screen a cDNA library in future.
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References
Birnboim, H. C. and Dolly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acid Res. 7, 1513-1523.
Blumberg, B., Jack Bolado, Jr., Fadila Derguini, A. Grey Craig, Tanya A. Moreno, Debabrata Chakravarti, Richard A. Heyman, Jochen Buck, and Ronald M. Evans (1996). Novel retinoic acid receptor ligands in Xenopus embryos. Proc. Natl. Acad. Sci. USA 93, 4873-4878.
Bouillet, P., Oulad-Abdelghani, M., Vicaire, S., Garnier, J., Schuhbaur, B., Dolle'. P. and Chambon, P. (1995). Efficient cloning of cDNAs of Retinoic Acid-responsive Genes in P19 Embryonal Carcinoma Cells and characterization of a Novel Mouse Gene, Stral (Mouse LERK-2/Eplg2). Dev. Biol. 170, 420-433.
Chen, Y., Huang, L., Rusco, A.F. and Solurch, M. (1992). Retinoic acid is enriched in Hensens node and is developmentally regulated in the early
chicken embryo. Proc. Natl. Acad. Sci. USA 89, 10056-10059.
Costaridis, P. Horton, C., Zeitlinger, J., Holder, N., and Maden, M. (1996). Endogenous retinoids in the zebrafish embryo and adult. Dev. Dyn. 205, 41-51.
Creech Kraft, J., Schuh, T., Juchau, M., and Kimelman, D. (1994). The retinoid X receptor ligand, 9-cis retinoic acid, is a potential regulator of early Xenopus development. Proc. Natl. Acad. Sci. USA. 91, 3067-3071.
Giovanna Alfano, Ivan Conte, Tiziana Caramico, Raffaella Avellino, Benedetta Pascal Dolle, Mathias W. Seeliger, and Sandro Banfi (2011).
Vax2 regulates retinoic acid distribution and cone opsin expression in the vertebrate eye. Development, 138, 261-271.
Hogan, B.L.M., Thaller, C. and Eichele, G. (1992). Evidence that Hensens node is a site of retinoic acid synthesis. Nature 359, 237-241.
Ina Strate, Tan H. Min, Dobromir lliev, and Edgar M. Pera (2009). Retinol dehydrogenase 10 is a feedback regulator of retinoic acid signaling during axis formation and patterning of the central nervous system. Development, 136, 461-472.
Jonk, L. J. C., De Jonge, M. E. J., Ver vaart, J. M. A., Wissink, S. and Kruijer, W. (1994). Isolation and Developmental Expression of Retinoic-Acid-Induced genes. Dev. Biol. 161, 604-614.
King, R. W. Jackson, P. K., Kirschner, M. W. (1994). Mitosis in transition. Cell. 79, 563-571.
Kirsten M. Spoorendonk, Josi Peterson-Maduro, Jorg Renn, Torsten Trowe, Sander Kranenbarg, Christoph Winkler, and Stefan Schulte-Merker (2008). Retinoic acid and Cyp26b1 are critical regulators of osteogenesis in the axial skeleton. Development, 135, 3765-3774.
LaRosa, G. J. and Gudas, L. J. (1988a). An early effect of retinoic acid: cloning of an mRNA (Era-1) exhibiting rapid and protein synthesis-independent induction during teratocarcinoma stem cell differentiation. Proc. Natl. Acad. Sci. USA 85, 329-333.
Lee, S. W., Tomasetto, C. and Sager, R. (1991). Positive Selection of candidate tumor-suppressor genes by subtractive hybridization. Proc. Natl. Acad. Sci. USA. 88, 2825-2829.
Liang, P., and Pardee, A. B. (1992). Differential Display of Eukaryotic Messenger RNA by Means of the Polymerase Chain Reaction. Science 257, 967-971.
Lisa L. Sandell, Brian W. Sanderson, Gennadiy Moiseyev, Teri Johnson, Kendra Young, Jean-Philippe Rey, Jian-xing Ma, Karen Staehling-Hampton, and Paul A. Trainor (2007). RDH10 is essential for synthesis of embryonic retinoic acid and is required for limb, craniofacial, and organ development. Genes Dev., 21, 1113-1124
Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982). Molecular cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N. Y.
Means, A. L. and Gudas, L. J. (1995). The Roles of Retinoids in Vertebrate Development. Annu. Rev. Biochem. 64, 201-233.
Muriel Rhinn and Pascal Dolle (2012). Retinoic acid signaling during development.
Development, 139, 843-858.
Naomi Moriya, shinji Komazaki, Shuji Takahashi, chika Yokota and Makoto Asashima (2000). In vitro pancreas formation from Xenopus ectoderm treated with activin and retinoic acid. Development, Growth and Differentiation. 42, 593-602.
Nieuwkoop, P. and Faber, J. (1967). Normal Table of Xenopus laevis (North-Holland, Amsterdam)
Panagiotis A. Tsonis, Melissa Tsavaris, Mindy K. Call, Roshantha A. S. Chandraratna, Katia Del Rio-Tsonis (2002). Expression and role of
retinoic acid receptor alpha in lens regeneration. Development, Growth and Differentiation. 44, 391-394.
Pearson, W. R. and Lipman, D. J. (1988). "Improved tools for biological sequence comparison". Proc. Natl. Acad. Sci. USA 84, 2444-2448.
Rosa A. Garcia-Fernandez, Claudia Perez-Martinez, Javier Espinosa Alvarez, Alex J. Duran Navarrete, Marria J. Garcia-Iglesias (2006). Mouse epidermal development effects of retinoic acid exposure in utero epidermal thickness. Verterinary Dermatology. 17, 36-44.
Sussie Dalvin, Katsumi Komatsuzaki, Mark A. Anselmo, David E. Kling, Jay J. Schnitzer, T. Bernard Kinane (2004). Retinoic acid decreases fetal lung mesenchymal cell proliferation in vivo and in vitro. Development, Growth and Differentiation. 46, 275-282.
Tsung-Chieh J. Wu, Lai Wang, Yu-Jui Y. Wan (2005). Differential Expression of Retinoic acid Receptor mRNA during Mouse
Embryogenesis Development Growth and Differentiation 34 685-691
You Katsuyama, Hidetoshi Saiga (2003). Retnoic acid affects patterning along the anterior-posterior axis of the ascidian embryo. Development, Growth and Differentiation. 40, 413-422.
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