Download PDF
Exp Clin Endocrinol Diabetes 2005; 113(1): 13-20
DOI: 10.1055/s-2004-830516
Article

J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York

Functional Thyrotropin Receptor Expression in the Pituitary Folliculo-Stellate Cell Line TtT/GF

L. J. S. Brokken1 , O. Bakker1 , W. M. Wiersinga1 , M. F. Prummel1
  • 1Department of Endocrinology and Metabolism, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands
Further Information

Publication History

Received: January 9, 2003 First decision: March 3, 2004

Accepted: May 13, 2004

Publication Date:
21 January 2005 (online)

Also available at
    Permissions and Reprints

Abstract

Thyrotropin secretion from the anterior pituitary is regulated mainly through TRH and thyroid hormones. Recent findings of a TSH receptor (TSHR) on folliculo-stellate (FS) cells in the human anterior pituitary indicate that TSH secretion might, in addition, be regulated in a paracrine manner via FS cells. In order to elucidate the physiological relevance of TSHR expression in FS cells we evaluated the effects of TSH on a murine FS cell line, TtT/GF. First, Western blot analysis confirmed the expression of TSHR protein in these cells. Second, three potential second messenger pathways were studied. Last, cDNA array hybridization was used to evaluate the effect of TSH on gene expression levels. TSH failed to induce either the adenylate cyclase/cAMP pathway, the phosphatidylinositol/calcium pathway, or the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) 3 pathway. Most of the genes regulated by TSH were related to cell proliferation, cell differentiation, and apoptosis. Moreover, TSH induced STAT5a and TGFβ2 expression. We report that TtT/GF cells express a functional TSHR that is not coupled to cAMP nor IP3 but probably signals through the JAK/STAT5a pathway. Functional TSHR expression in this cell line offers an in vitro model to study the role of TSHR in FS cells.

Key words

Thyrotropin receptor - folliculo-stellate cells - cell line - TtT/GF

References

  • 1 Allaerts W, Carmeliet P, Denef C. New perspectives in the function of pituitary folliculo-stellate cells.  Mol Cell Endocrinol. 1990;  71 73-81
    CrossrefPubMedGoogle Scholar
  • 2 Brokken L JS, Scheenhart J W, Wiersinga W M, Prummel M F. Suppression of serum TSH by Graves' Ig: evidence for a functional pituitary TSH receptor.  J Clin Endocrinol Metabol. 2001;  86 4814-4817
    CrossrefPubMedGoogle Scholar
  • 3 Brokken L JS, Leendertse M, Bakker O, Wiersinga W M, Prummel M F. Expression of adenohypophyseal-hormone receptors in a murine folliculo-stellate cell line.  Hormone Metabol Res. 2004;  36 538-541
    PubMedGoogle Scholar
  • 4 Chazenbalk G D, Kakinuma A, Jaume J C, McLachlan S M, Rapoport B. Evidence for negative cooperativity among human thyrotropin receptors overexpressed in mammalian cells.  Endocrinol. 1996;  137 4586-4591
    CrossrefPubMedGoogle Scholar
  • 5 Chuang C C, Tan S K, Tai L K, Hsin J P, Wang F F. Evidence for the involvement of protein kinase C in the inhibition of prolactin gene expression by transforming growth factor-beta2.  Mol Pharmacol. 1998;  53 1054-1061
    PubMedGoogle Scholar
  • 6 De Groot R P, Raaijmakers J A, Lammers J W, Koenderman L. STAT5-dependent cyclinD1 and Bcl-xL expression in Bcr-Abl-transformed cells.  Mol Cel Biol Res Comm. 2000;  3 299-305
    PubMedGoogle Scholar
  • 7 Ferrara N, Schweigerer L, Neufeld G, Mitchell R, Gospodarowicz D. Pituitary follicular cells produce basic fibroblast growth factor.  Proc Nat Acad Sci USA. 1987;  84 5773-5777
    PubMedGoogle Scholar
  • 8 Ferrara N, Henzel W J. Pituitary follicular cells secrete a novel heparin-binding growth factor specific for vascular endothelial cells.  Biochem Biophys Res Comm. 1989;  161 851-858
    CrossrefPubMedGoogle Scholar
  • 9 Ferrara N, Winer J, Henzel W J. Pituitary follicular cells secrete an inhibitor of aortic endothelial cell growth: identification as leukemia inhibitory factor.  Proc Nat Acad Sci USA. 1992;  89 698-702
    PubMedGoogle Scholar
  • 10 Field J B, Ealey P A, Marshall N J, Cockcroft S. Thyroid-stimulating hormone stimulates increases in inositol phosphates as well as cyclic AMP in the FRTL-5 rat thyroid cell line.  Biochem J. 1987;  247 519-524
    PubMedGoogle Scholar
  • 11 Gospodarowicz D, Lau K. Pituitary follicular cells secrete both vascular endothelial growth factor and follistatin.  Biochem Biophys Res Comm. 1989;  165 292-298
    PubMedGoogle Scholar
  • 12 Grandison L, Nolan G P, Pfaff D W. Activation of the transcription factor NF-KB in GH3 pituitary cells.  Mol Cell Endocrinol. 1994;  106 9-15
    CrossrefPubMedGoogle Scholar
  • 13 Inoue K, Matsumoto H, Koyama C, Shibata K, Nakazato Y, Ito A. Establishment of a folliculo-stellate-like cell line from a murine thyrotropic pituitary tumor.  Endocrinol. 1992;  131 3110-3116
    CrossrefPubMedGoogle Scholar
  • 14 Inoue K, Couch E F, Takano K, Ogawa S. The structure and function of folliculo-stellate cells in the anterior pituitary gland.  Arch Histol Cytol. 1999;  62 205-218
    PubMedGoogle Scholar
  • 15 Jin L, Qian X, Kulig E, Sanno N, Scheithauer B W, Kovacs K, Young Jr W F, Lloyd R V. Transforming growth factor-beta, transforming growth factor-beta receptor II, and p27Kip1 expression in nontumorous and neoplastic human pituitaries.  Am J Pathol. 1997;  151 509-519
    PubMedGoogle Scholar
  • 16 Jin L, Tsumanuma I, Ruebel K H, Bayliss J M, Lloyd R V. Analysis of homogeneous populations of anterior pituitary folliculostellate cells by laser capture microdissection and reverse transcription-polymerase chain reaction.  Endocrinol. 2001;  142 1703-1709
    CrossrefPubMedGoogle Scholar
  • 17 Laurent E, Mockel J, Van Sande J, Graff I, Dumont J E. Dual activation by thyrotropin of the phospholipase C and cyclic AMP cascades in human thyroid.  Mol Cel Endocrinol. 1987;  52 273-278
    PubMedGoogle Scholar
  • 18 Nicholson L B, Vlase H, Graves P, Nilsson M, Molne J, Huang G C, Morgenthaler N G, Davies T F, McGregor A M, Banga J P. Monoclonal antibodies to the human TSH receptor: epitope mapping and binding to the native receptor on the basolateral plasma membrane of thyroid follicular cells.  J Mol Endocrinol. 1996;  16 159-170
    PubMedGoogle Scholar
  • 19 Nishikawa T, Yamashita S, Namba H, Usa T, Tominaga T, Kimura H, Izumi M, Nagataki S. Interferon-gamma inhibition of human thyrotropin receptor gene expression.  J Clin Endocrinol Metabol. 1993;  77 1084-1089
    CrossrefPubMedGoogle Scholar
  • 20 Park E S, Kim H, Suh J M, Park S J, You S H, Chung H K, Lee K W, Kwon O Y, Cho B Y, Kim Y K, Ro H K, Chung J, Shong M. Involvement of JAK/STAT (Janus kinase/signal transducer and activator of transcription) in the thyrotropin signaling pathway.  Mol Endocrinol. 2000;  14 662-670
    CrossrefPubMedGoogle Scholar
  • 21 Perret J, Ludgate M, Libert F, Gerard C, Dumont J E, Vassart G, Parmentier M. Stable expression of the human TSH receptor in CHO cells and characterization of differentially expressing clones.  Biochem Biophys Res Comm. 1990;  171 1044-1050
    CrossrefPubMedGoogle Scholar
  • 22 Philp N J, Grollman E F. Thyrotropin and norepinephrine stimulate the metabolism of phosphoinositides in FRTL-5 thyroid cells.  Fed Eur Biochem Soc Lett. 1986;  202 193-196
    PubMedGoogle Scholar
  • 23 Piotrowska U, Adler G, Gardas A, Gietka-Czernel M, Kaniewski M, Banga J P. Cross-reactivity of a monoclonal antibody to the aminoterminal region of thyrotropin receptor with the serum protein α1-antitrypsin.  Thyroid. 2002;  12 563-570
    CrossrefPubMedGoogle Scholar
  • 24 Prummel M F, Brokken L JS, Meduri G, Misrahi M, Bakker O, Wiersinga W M. Expression of the thyroid-stimulating hormone receptor in the folliculo-stellate cells of the human anterior pituitary.  J Clin Endocrinol Metabol. 2000;  85 4347-4353
    CrossrefPubMedGoogle Scholar
  • 25 Renner U, Gloddek J, Arzt E, Inoue K, Stalla G K. Interleukin-6 is an autocrine growth factor for folliculostellate-like TtT/GF mouse pituitary tumor cells.  Exp Clin Endocrinol Diabetes. 1997;  105 345-352
    PubMedGoogle Scholar
  • 26 Renner U, Lohrer P, Schaaf L, Feirer M, Schmitt K, Onofri C, Arzt E, Stalla G K. Transforming growth factor-β stimulates vascular endothelial growth factor production by folliculo-stellate pituitary cells.  Endocrinol. 2002;  143 3759-3765
    CrossrefPubMedGoogle Scholar
  • 27 Rinehart J F, Farquhar M G. Electron microscopic study of the anterior pituitary gland.  J Histochem Cytochem. 1953;  1 93-113
    PubMedGoogle Scholar
  • 28 Russo D, Chazenbalk G D, Nagayama Y, Wadsworth H L, Seto P, Rapoport B. A new structural model for the thyrotropin (TSH) receptor, as determined by covalent cross-linking of TSH to the recombinant receptor in intact cells: evidence for a single polypeptide chain.  Mol Endocrinol. 1991;  5 1607-1612
    PubMedGoogle Scholar
  • 29 Saunier B, Pierre M, Jacquemin C, Courtin F. Evidence for cAMP-independent thyrotropin effects on astroglial cells.  Eur J Biochem. 1993;  218 1091-1094
    CrossrefPubMedGoogle Scholar
  • 30 Socolovsky M, Fallon A E, Wang S, Brugnara C, Lodish H F. Fetal anemia and apoptosis of red cell progenitors in Stat5a-/-5 b-/- mice: a direct role for Stat5 in Bcl-X(L) induction.  Cell. 1999;  98 181-191
    CrossrefPubMedGoogle Scholar
  • 31 Theodoropoulou M, Arzberger T, Gruebler Y, Korali Z, Mortini P, Joba W, Heufelder A E, Stalla G K, Schaaf L. Thyrotrophin receptor protein expression in normal and adenomatous human pituitary.  J Endocrinol. 2000;  167 7-13
    CrossrefPubMedGoogle Scholar
  • 32 Tournier C, Gavaret J M, Jacquemin C, Pierre M, Saunier B. Stimulation of mitogen-activated protein kinase by thyrotropin in astrocytes.  European J of Biochemistry. 1995;  228 16-22
    PubMedGoogle Scholar
  • 33 Vankelecom H, Andries M, Billiau A, Denef C. Evidence that folliculo-stellate cells mediate the inhibitory effect of interferon-gamma on hormone secretion in rat anterior pituitary cell cultures.  Endocrinol. 1992;  130 3537-3546
    CrossrefPubMedGoogle Scholar
  • 34 Wolff J, Jones A B. The purification of bovine thyroid plasma membranes and the properties of membrane-bound adenyl cyclase.  J Biol Chem. 1971;  246 3939-3947
    PubMedGoogle Scholar
  • 35 Yamashita K, Field J B. Preparation of thyroid plasma membranes containing a TSH-responsive adenyl cyclase.  Biochem Biophys Res Comm. 1970;  40 171-178
    CrossrefPubMedGoogle Scholar
  • 36 Yu C L, Meyer D J, Campbell G S, Larner A C, Carter-Su C, Schwartz J, Jove R. Enhanced DNA-binding activity of a Stat3-related protein in cells transformed by the Src oncoprotein.  Science. 1995;  269 81-83
    PubMedGoogle Scholar

Leon J. S. Brokken

Department of Physiology
University of Turku

Kiinamyllynkatu 10

20520 Turku

Finland

Phone: + 35823337376

Fax: + 35 8 32 50 26 10

Email: Leon.Brokken@utu.fi

      >