This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Liu, Y.-Y. Articles by Brent, G. A. Search for Related Content PubMed PubMed Citation Articles by Liu, Y.-Y. Articles by Brent, G. A.

A Targeted Thyroid Hormone Receptor Gene Dominant-Negative Mutation (P398H) Selectively Impairs Gene Expression in Differentiated Embryonic Stem Cells

Molecular Endocrinology Laboratory and Research Service, Veterans Affairs Greater Los Angeles Healthcare System, Departments of Medicine and Physiology, University of California Los Angeles School of Medicine, Los Angeles, California 90073

Address all correspondence and requests for reprints to: Gregory A. Brent, Molecular Endocrinology Laboratory, Veterans Affairs Greater Los Angeles Healthcare System, Building 114, Room 230, 11301 Wilshire Boulevard, Los Angeles, California 90073. E-mail: . gbrent{at}ucla.edu

Thyroid hormone and retinoic acid (RA) are essential for normal neural development in vivo, yet all in vitro differentiation strategies of embryonic stem (ES) cells use only RA. We developed a novel differentiation strategy of mouse ES cells using T₃. A dominant-negative knock-in point mutation (P398H) was introduced into the thyroid hormone receptor gene to determine the influence of T₃ on ES cell differentiation. Differentiation promoted by T₃ (1 nM), RA (1 µM), or combined T₃/RA was assessed in wild-type (wt) and mutant (m) ES cells on the basis of neuronal-specific gene expression and cell cycle. T₃ alone stimulated neural differentiation in a similar fashion as that seen with RA in both wtES and mES cells. Expression of neurogranin and Ca²⁺/calmodulin-dependent kinase IV mRNA (identified in vivo as T₃-regulated genes), however, was markedly reduced in mES, compared with wtES cells. RA treatment enhanced apoptosis, significantly greater than that seen with T₃ stimulation. T₃ treatment given with RA significantly reduced the apoptotic effects of RA, an effect not seen in mES cells. T₃-induced ES cell neural differentiation of thyroid hormone mutant and wtES cells provides an in vitro model to study T₃-dependent gene regulation in neural development. This system could also be used to identify novel T₃-regulated genes. The modulation of the apoptotic effects of RA by T₃ may have implications for stem cell therapy.

This article has been cited by other articles:

				L. Quignodon, S. Vincent, H. Winter, J. Samarut, and F. Flamant A Point Mutation in the Activation Function 2 Domain of Thyroid Hormone Receptor {alpha}1 Expressed after CRE-Mediated Recombination Partially Recapitulates Hypothyroidism Mol. Endocrinol., October 1, 2007; 21(10): 2350 - 2360. [Abstract] [Full Text] [PDF]

				I. Jones, L. Ng, H. Liu, and D. Forrest An Intron Control Region Differentially Regulates Expression of Thyroid Hormone Receptor {beta}2 in the Cochlea, Pituitary, and Cone Photoreceptors Mol. Endocrinol., May 1, 2007; 21(5): 1108 - 1119. [Abstract] [Full Text] [PDF]

				Y.-Y. Liu, R. S. Heymann, F. Moatamed, J. J. Schultz, D. Sobel, and G. A. Brent A Mutant Thyroid Hormone Receptor {alpha} Antagonizes Peroxisome Proliferator-Activated Receptor {alpha} Signaling in Vivo and Impairs Fatty Acid Oxidation Endocrinology, March 1, 2007; 148(3): 1206 - 1217. [Abstract] [Full Text] [PDF]

				F. Flamant, K. Gauthier, and J. Samarut Thyroid Hormones Signaling Is Getting More Complex: STORMs Are Coming Mol. Endocrinol., February 1, 2007; 21(2): 321 - 333. [Abstract] [Full Text] [PDF]

				M. Nygard, N. Becker, B. Demeneix, K. Pettersson, and M. Bondesson Thyroid hormone-mediated negative transcriptional regulation of Necdin expression. J. Mol. Endocrinol., June 1, 2006; 36(3): 517 - 530. [Abstract] [Full Text] [PDF]

				Y.-Y. Liu and G. A. Brent Thyroid Hormone-Dependent Gene Expression in Differentiated Embryonic Stem Cells and Embryonal Carcinoma Cells: Identification of Novel Thyroid Hormone Target Genes by Deoxyribonucleic Acid Microarray Analysis Endocrinology, February 1, 2005; 146(2): 776 - 783. [Abstract] [Full Text] [PDF]

				M. Fernandez, A. Giuliani, S. Pirondi, G. D'Intino, L. Giardino, L. Aloe, R. Levi-Montalcini, and L. Calza Thyroid hormone administration enhances remyelination in chronic demyelinating inflammatory disease PNAS, November 16, 2004; 101(46): 16363 - 16368. [Abstract] [Full Text] [PDF]

				D. Forrest The Developing Brain and Maternal Thyroid Hormone: Finding the Links Endocrinology, September 1, 2004; 145(9): 4034 - 4036. [Full Text] [PDF]

				Y.-Y. Liu, J. J. Schultz, and G. A. Brent A Thyroid Hormone Receptor {alpha} Gene Mutation (P398H) Is Associated with Visceral Adiposity and Impaired Catecholamine-stimulated Lipolysis in Mice J. Biol. Chem., October 3, 2003; 278(40): 38913 - 38920. [Abstract] [Full Text] [PDF]

				Y.-Y. Liu and G. A. Brent A Complex Deoxyribonucleic Acid Response Element in the Rat Ca2+/Calmodulin-Dependent Protein Kinase IV Gene 5'-Flanking Region Mediates Thyroid Hormone Induction and Chicken Ovalbumin Upstream Promoter Transcription Factor 1 Repression Mol. Endocrinol., November 1, 2002; 16(11): 2439 - 2451. [Abstract] [Full Text] [PDF]

				D. Forrest Editorial: Twists in the Tail--Change-of-Function Mutations in Thyroid Hormone Receptors Endocrinology, July 1, 2002; 143(7): 2466 - 2468. [Full Text] [PDF]