The location and the regulation of the type I-iodothyronine 5′-monodeiodinase (type I-MD) in the rat thyroid: studies using a specific anti-type I-MD antibody

Ferruccio Santini, Luca Chiovato, Paola Lapi, Mario Lupetti, Amelio Dolfi, Francesco Bianchi, Nunzia Bernardini, Giovanna Bendinelli, Claudia Mammoli, Paolo Vitti, Inder J. Chopra, Aldo Pinchera

Research output: Contribution to journalArticle

Abstract

Type I-iodothyronine monodeiodinase (type I-MD) is abundant in thyroid tissue and contributes to the generation of T3 secreted by the gland. The availability of a specific antibody against rat type I-MD (type I-MD Ab) allowed us to directly identify this enzyme in rat thyroid glands, and in a differentiated strain of rat thyroid cells maintained in continuous culture (FRTL-5 cells). FRTL-5 cells maintain many differentiated functions of thyroid cells, including the expression of TSH receptor and thyroid peroxidase. Using an immunohistochemical technique on rat thyroid sections, a clear staining for type I-MD was demonstrated in follicular cells. The degree of immunoreactivity was greater in small follicles containing little amounts of colloid compared to large follicles lined by functionally inactive cells. Using immunofluorescence (IFL), a strong staining for type I-MD was observed in FRTL-5 cells grown in medium containing TSH. Both in vivo and in culture the staining for type I-MD was localised in the cytoplasm of thyroid cells, while nuclei were negative. Interestingly, no surface staining was shown when viable FRTL-5 cells were submitted to the same IFL procedure. TSH deprivation for 7 days was followed by the disappearance of type I-MD. Immunoreactivity for type I-MD was recovered by addition of TSH, forskolin or thyroid stimulating antibody (TSAb) to TSH-deprived FRTL-5 cells. The effect of TSH was prevented by cycloheximide. There was no induction of type I-MD when IGF-I was added to FRTL-5 cells. In conclusion, our results indicate that: (i) type I-MD is abundant in intracytoplasmic membranes of rat thyroid cells, both in vivo and in culture, while it is not expressed on the outer side of the plasma membrane; (ii) the synthesis of type I-MD is TSH-dependent through pathways that involve cAMP production, while it is not influenced by IGF-I which activates the tyrosine protein kinase pathway; (iii) TSAb mimics TSH in inducing the expression of type I-MD in FRTL-5 cells, and may thus influence the intrathyroidal generation of T3 in patients with Graves' disease.

Original languageEnglish
Pages (from-to)195-203
Number of pages9
JournalMolecular and Cellular Endocrinology
Volume110
Issue number1-2
DOIs
Publication statusPublished - Apr 28 1995

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Iodide Peroxidase
Rats
Thyroid Gland
Antibodies
Thyroid-Stimulating Immunoglobulins
Staining and Labeling
Insulin-Like Growth Factor I
Fluorescent Antibody Technique
Thyrotropin Receptors
Graves Disease
Colloids
Colforsin
Cell membranes
Cycloheximide
Cell Nucleus
Protein-Tyrosine Kinases
Cytoplasm

Keywords

  • Graves' disease
  • IGF I
  • Thyroid hormone
  • Thyroid stimulating antibody
  • Type I-iodothyronine monodeiodinase

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

The location and the regulation of the type I-iodothyronine 5′-monodeiodinase (type I-MD) in the rat thyroid : studies using a specific anti-type I-MD antibody. / Santini, Ferruccio; Chiovato, Luca; Lapi, Paola; Lupetti, Mario; Dolfi, Amelio; Bianchi, Francesco; Bernardini, Nunzia; Bendinelli, Giovanna; Mammoli, Claudia; Vitti, Paolo; Chopra, Inder J.; Pinchera, Aldo.

In: Molecular and Cellular Endocrinology, Vol. 110, No. 1-2, 28.04.1995, p. 195-203.

Research output: Contribution to journalArticle

Santini, F, Chiovato, L, Lapi, P, Lupetti, M, Dolfi, A, Bianchi, F, Bernardini, N, Bendinelli, G, Mammoli, C, Vitti, P, Chopra, IJ & Pinchera, A 1995, 'The location and the regulation of the type I-iodothyronine 5′-monodeiodinase (type I-MD) in the rat thyroid: studies using a specific anti-type I-MD antibody', Molecular and Cellular Endocrinology, vol. 110, no. 1-2, pp. 195-203. https://doi.org/10.1016/0303-7207(95)03532-C
Santini, Ferruccio ; Chiovato, Luca ; Lapi, Paola ; Lupetti, Mario ; Dolfi, Amelio ; Bianchi, Francesco ; Bernardini, Nunzia ; Bendinelli, Giovanna ; Mammoli, Claudia ; Vitti, Paolo ; Chopra, Inder J. ; Pinchera, Aldo. / The location and the regulation of the type I-iodothyronine 5′-monodeiodinase (type I-MD) in the rat thyroid : studies using a specific anti-type I-MD antibody. In: Molecular and Cellular Endocrinology. 1995 ; Vol. 110, No. 1-2. pp. 195-203.
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AU - Chiovato, Luca

AU - Lapi, Paola

AU - Lupetti, Mario

AU - Dolfi, Amelio

AU - Bianchi, Francesco

AU - Bernardini, Nunzia

AU - Bendinelli, Giovanna

AU - Mammoli, Claudia

AU - Vitti, Paolo

AU - Chopra, Inder J.

AU - Pinchera, Aldo

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N2 - Type I-iodothyronine monodeiodinase (type I-MD) is abundant in thyroid tissue and contributes to the generation of T3 secreted by the gland. The availability of a specific antibody against rat type I-MD (type I-MD Ab) allowed us to directly identify this enzyme in rat thyroid glands, and in a differentiated strain of rat thyroid cells maintained in continuous culture (FRTL-5 cells). FRTL-5 cells maintain many differentiated functions of thyroid cells, including the expression of TSH receptor and thyroid peroxidase. Using an immunohistochemical technique on rat thyroid sections, a clear staining for type I-MD was demonstrated in follicular cells. The degree of immunoreactivity was greater in small follicles containing little amounts of colloid compared to large follicles lined by functionally inactive cells. Using immunofluorescence (IFL), a strong staining for type I-MD was observed in FRTL-5 cells grown in medium containing TSH. Both in vivo and in culture the staining for type I-MD was localised in the cytoplasm of thyroid cells, while nuclei were negative. Interestingly, no surface staining was shown when viable FRTL-5 cells were submitted to the same IFL procedure. TSH deprivation for 7 days was followed by the disappearance of type I-MD. Immunoreactivity for type I-MD was recovered by addition of TSH, forskolin or thyroid stimulating antibody (TSAb) to TSH-deprived FRTL-5 cells. The effect of TSH was prevented by cycloheximide. There was no induction of type I-MD when IGF-I was added to FRTL-5 cells. In conclusion, our results indicate that: (i) type I-MD is abundant in intracytoplasmic membranes of rat thyroid cells, both in vivo and in culture, while it is not expressed on the outer side of the plasma membrane; (ii) the synthesis of type I-MD is TSH-dependent through pathways that involve cAMP production, while it is not influenced by IGF-I which activates the tyrosine protein kinase pathway; (iii) TSAb mimics TSH in inducing the expression of type I-MD in FRTL-5 cells, and may thus influence the intrathyroidal generation of T3 in patients with Graves' disease.

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