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

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 T(3). A dominant-negative knock-in point mutation (P398H) was introduced into the thyroid hormone receptor alpha gene to determine the influence of T(3) on ES cell differentiation. Differentiation promoted by T(3) (1 nM), RA (1 microM), or combined T(3)/RA was assessed in wild-type (wt) and mutant (m) ES cells on the basis of neuronal-specific gene expression and cell cycle. T(3) alone stimulated neural differentiation in a similar fashion as that seen with RA in both wtES and mES cells. Expression of neurogranin and Ca(2+)/calmodulin-dependent kinase IV mRNA (identified in vivo as T(3)-regulated genes), however, was markedly reduced in mES, compared with wtES cells. RA treatment enhanced apoptosis, significantly greater than that seen with T(3) stimulation. T(3) treatment given with RA significantly reduced the apoptotic effects of RA, an effect not seen in mES cells. T(3)-induced ES cell neural differentiation of thyroid hormone alpha mutant and wtES cells provides an in vitro model to study T(3)-dependent gene regulation in neural development. This system could also be used to identify novel T(3)-regulated genes. The modulation of the apoptotic effects of RA by T(3) may have implications for stem cell therapy.

[1]  L.,et al.  Role of retinoic acid and oxidative stress in embryonic stem cell death and neuronal differentiation , 2003 .

[2]  D. Yamaguchi,et al.  Extracellular matrix alters the relationship between tritiated thymidine incorporation and proliferation of MC3T3‐E1 cells during osteogenesis in vitro , 2002, Cell proliferation.

[3]  橋本 貢士.  An unliganded thyroid hormone receptor causes severe neurological dysfunction , 2002 .

[4]  M. Willingham,et al.  A targeted dominant negative mutation of the thyroid hormone α1 receptor causes increased mortality, infertility, and dwarfism in mice , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[5]  L Gepstein,et al.  Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes. , 2001, The Journal of clinical investigation.

[6]  B. Fleischmann,et al.  Indispensable tools: embryonic stem cells yield insights into the human heart. , 2001, The Journal of clinical investigation.

[7]  L. Sokoloff,et al.  Brain glucose utilization in mice with a targeted mutation in the thyroid hormone α or β receptor gene , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Willingham,et al.  Mice with a targeted mutation in the thyroid hormone beta receptor gene exhibit impaired growth and resistance to thyroid hormone. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[9]  D. Holtzman,et al.  Impaired Synaptic Plasticity and cAMP Response Element-Binding Protein Activation in Ca2+/Calmodulin-Dependent Protein Kinase Type IV/Gr-Deficient Mice , 2000, The Journal of Neuroscience.

[10]  M. McCarthy,et al.  Neurotoxic effects of endocrine disruptors. , 1999, Current opinion in neurology.

[11]  D. Forrest,et al.  Mice devoid of all known thyroid hormone receptors are viable but exhibit disorders of the pituitary-thyroid axis, growth, and bone maturation. , 1999, Genes & development.

[12]  J. Samarut,et al.  Different functions for the thyroid hormone receptors TRα and TRβ in the control of thyroid hormone production and post‐natal development , 1999 .

[13]  J G Hollowell,et al.  Maternal thyroid deficiency during pregnancy and subsequent neuropsychological development of the child. , 1999, The New England journal of medicine.

[14]  C. Babinet,et al.  Cerebellar defect and impaired motor coordination in mice lacking vimentin , 1999, Glia.

[15]  Peter J. Donovan,et al.  Derivation of pluripotent stem cells from cultured human primordial germ cells , 1998 .

[16]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[17]  H. Beug,et al.  The thyroid hormone receptor functions as a ligand‐operated developmental switch between proliferation and differentiation of erythroid progenitors , 1998, The EMBO journal.

[18]  J. Samarut,et al.  Involvement of thyroid hormone and its alpha receptor in avian neurulation. , 1998, Developmental biology.

[19]  G. Brent,et al.  Thyroid Hormone Receptor Gene Knockouts , 1998, Trends in Endocrinology & Metabolism.

[20]  Y. Su,et al.  Auto-regulation of thyroid hormone receptor genes during metamorphosis: roles in apoptosis and cell proliferation. , 1998, The International journal of developmental biology.

[21]  D. Forrest,et al.  Beta receptor isoforms are not essential for thyroid hormone-dependent acceleration of PCP-2 and myelin basic protein gene expression in the developing brains of neonatal mice , 1998, Molecular and Cellular Endocrinology.

[22]  J. Oppenheimer,et al.  Molecular basis of thyroid hormone-dependent brain development. , 1997, Endocrine reviews.

[23]  C. Dehay,et al.  The T3Rα gene encoding a thyroid hormone receptor is essential for post‐natal development and thyroid hormone production , 1997, The EMBO journal.

[24]  P. Chambon,et al.  RXRalpha‐null F9 embryonal carcinoma cells are resistant to the differentiation, anti‐proliferative and apoptotic effects of retinoids. , 1996, The EMBO journal.

[25]  D. Forrest,et al.  Thyroid hormone receptor β is essential for development of auditory function , 1996, Nature Genetics.

[26]  S. Orkin,et al.  In vitro differentiation of murine embryonic stem cells. New approaches to old problems. , 1996, The Journal of clinical investigation.

[27]  B. Wiedenmann,et al.  Differentiation of pluripotent embryonic stem cells into the neuronal lineage in vitro gives rise to mature inhibitory and excitatory neurons , 1995, Mechanisms of Development.

[28]  V. M. Neto,et al.  Thyroid hormone and conditioned medium effects on astroglial cells from hypothyroid and normal rat brain: Factor secretion, cell differentiation, and proliferation , 1995, Journal of neuroscience research.

[29]  D. Gottlieb,et al.  Embryonic stem cells express neuronal properties in vitro. , 1995, Developmental biology.

[30]  G. Delong,et al.  Timing of vulnerability of the brain to iodine deficiency in endemic cretinism. , 1994, The New England journal of medicine.

[31]  P. Yen,et al.  Differential expression of thyroid hormone receptor isoforms in neurons and astroglial cells. , 1994, Endocrinology.

[32]  L. Gudas.  Retinoids and vertebrate development. , 1994, The Journal of biological chemistry.

[33]  G. Brent,et al.  Thyroid hormone receptor-alpha inhibits retinoic acid-responsive gene expression and modulates retinoic acid-stimulated neural differentiation in mouse embryonic stem cells. , 1994, Molecular endocrinology.

[34]  J. Puymirat,et al.  Overexpression of the beta 1 thyroid receptor induces differentiation in neuro-2a cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Harney,et al.  Dominant negative inhibition by mutant thyroid hormone receptors is thyroid hormone response element and receptor isoform specific. , 1993, Molecular endocrinology.

[36]  S. Boyages.  Clinical review 49: Iodine deficiency disorders. , 1993, The Journal of clinical endocrinology and metabolism.

[37]  B. Seliger,et al.  Thyroid hormone receptor/c-erbA: control of commitment and differentiation in the neuronal/chromaffin progenitor line PC12 , 1993, The Journal of cell biology.

[38]  R. Mains,et al.  COOH-terminal signals mediate the trafficking of a peptide processing enzyme in endocrine cells , 1993, The Journal of cell biology.

[39]  S. P. Porterfield,et al.  The role of thyroid hormones in prenatal and neonatal neurological development--current perspectives. , 1993, Endocrine reviews.

[40]  L. Goya,et al.  Alterations in the growth and protein content of human neuroblastoma cells in vitro induced by thyroid hormones, stress and ageing. , 1993, Journal of reproduction and fertility. Supplement.

[41]  M. Koga,et al.  Effects of retinoic acid on estrogen- and thyroid hormone-induced growth in a newly established rat pituitary tumor cell line , 1992, The Journal of Steroid Biochemistry and Molecular Biology.

[42]  J. Bernal,et al.  Adult rat brain is sensitive to thyroid hormone. Regulation of RC3/neurogranin mRNA. , 1992, The Journal of clinical investigation.

[43]  M Maden,et al.  Retinoic acid and development of the central nervous system , 1992, BioEssays : news and reviews in molecular, cellular and developmental biology.

[44]  W. Young,et al.  Spatial and temporal expression of alpha- and beta-thyroid hormone receptor mRNAs, including the beta 2-subtype, in the developing mammalian nervous system , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[45]  David A. Jones,et al.  cDNA sequence and differential expression of the mouse Ca2+/calmodulin‐dependent protein kinase IV gene , 1991, FEBS letters.

[46]  J. Bernal,et al.  Independent Expression of the α and β c-erbA Genes in Developing Rat Brain , 1991 .

[47]  Allan Bradley,et al.  Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice , 1991, Cell.

[48]  D. Forrest,et al.  Distinct functions for thyroid hormone receptors alpha and beta in brain development indicated by differential expression of receptor genes. , 1991, The EMBO journal.

[49]  T. Doetschman,et al.  Gene targeting in embryonic stem cells. , 1991, Biotechnology.

[50]  R. Krumlauf,et al.  Introduction of a subtle mutation into the Hox-2.6 locus in embryonic stem cells , 1991, Nature.

[51]  Y. Ben-Ari,et al.  Neurogranin: immunocytochemical localization of a brain-specific protein kinase C substrate , 1990, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[52]  F. Bloom,et al.  Subtractive cDNA cloning of RC3, a rodent cortex‐enriched mRNA encoding a novel 78 residue protein , 1990, Journal of neuroscience research.

[53]  H. Weintraub,et al.  Identification of a retinoic acid-sensitive period during primary axis formation in Xenopus laevis. , 1990, Genes & development.

[54]  D. Forrest,et al.  Contrasting developmental and tissue‐specific expression of alpha and beta thyroid hormone receptor genes. , 1990, The EMBO journal.

[55]  J. de Vijlder,et al.  Maternal-fetal transfer of thyroxine in congenital hypothyroidism due to a total organification defect or thyroid agenesis. , 1990, The New England journal of medicine.

[56]  J. Harney,et al.  Thyroid hormone aporeceptor represses T3-inducible promoters and blocks activity of the retinoic acid receptor. , 1989, The New biologist.

[57]  P. Timiras,et al.  Thyroid hormones and nervous system development. , 1989, Biology of the neonate.

[58]  J. de Vellis,et al.  Regulation of neuroblast proliferation by hormones and growth factors in chemically defined medium , 1988, Journal of cellular physiology.

[59]  Y. Yamada,et al.  Structure of the amino-terminal portion of the murine alpha 1(IV) collagen chain and the corresponding region of the gene. , 1988, The Journal of biological chemistry.

[60]  S. Hockfield,et al.  Identification of major cell classes in the developing mammalian nervous system , 1985, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[61]  D. Fisher.  Thyroid hormone effects on growth and development , 1985 .

[62]  J. Yamakawa,et al.  Filter-binding assay procedure for thyroid hormone receptors. , 1983, Analytical biochemistry.