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. 1965 Jul 1;26(1):177–185. doi: 10.1083/jcb.26.1.177

THE INTRACELLULAR LOCALIZATION OF PITUITARY THYROTROPIC HORMONE

Francis S Greenspan 1, Judy R Hargadine 1
PMCID: PMC2106704  PMID: 5323607

Abstract

The intracellular localization of a bovine anterior pituitary preparation of thyroid-stimulating hormone (TSH) was studied in guinea pigs and dogs. The preparation was administered intravascularly or applied directly to tissue sections. TSH was detected by an indirect technique utilizing bovine TSH antiserum and fluorescein-labeled anti-rabbit globulin; the presence of TSH in the tissue was indicated by fluorescence when the tissue was examined under the microscope with an ultraviolet light source. After either intravascular administration or direct application of the TSH preparation, striking fluorescence was found in the nuclei of the thyroid cells and to a lesser degree in the nuclei of retro-orbital fat tissue and kidney tubules in both species studied. A little fluorescence was also seen in spleen tissue. No fluorescence was noted in comparable tissues removed from control animals injected with bovine albumin or globulin or when the tissues were treated with the fluorescein-labeled globulin alone. Fluorescence was also noted in the nuclei of adrenal cells treated with unabsorbed antiserum, but this was greatly diminished when antiserum absorbed with crystalline ACTH was used. The positive reactions were all markedly decreased when the tissues were treated with antisera absorbed with the original TSH preparation. Fluorescence was noted in the cytoplasm of pituitary tissue from both treated and control animals, suggesting a cross-reaction between the bovine pituitary antisera and guinea pig or dog hypophysis. The indirect technique seems to be highly satisfactory for demonstration of the pitiutary hormone within the cell. In addition, the demonstration of immunologically active anterior pituitary TSH bound to cell nuclei offers a clue to the site of action of this hormone.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ASCH L., ARON C. [Research on the role of the kidney in the catabolism of thyrostimulin]. C R Seances Soc Biol Fil. 1962;156:1693–1694. [PubMed] [Google Scholar]
  2. COONS A. H., KAPLAN M. H. Localization of antigen in tissue cells; improvements in a method for the detection of antigen by means of fluorescent antibody. J Exp Med. 1950 Jan 1;91(1):1–13. doi: 10.1084/jem.91.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COONS A. H., LEDUC E. H., KAPLAN M. H. Localization of antigen in tissue cells. VI. The fate of injected foreign proteins in the mouse. J Exp Med. 1951 Feb;93(2):173–188. doi: 10.1084/jem.93.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. CRUICKSHANK B., CURRIE A. R. Localization of tissue antigens with the fluorescent antibody technique: application to human anterior pituitary hormones. Immunology. 1958 Jan;1(1):13–26. [PMC free article] [PubMed] [Google Scholar]
  5. FERGUSON J. J., Jr HORMONES AND MACROMOLECULES. Ann Intern Med. 1964 May;60:925–928. doi: 10.7326/0003-4819-60-5-925. [DOI] [PubMed] [Google Scholar]
  6. FREINKEL N. Extrathyroidal actions of pituitary thyrotropin: effects on the carbohydrate, lipid and respiratory metabolism of rat adipose tissue. J Clin Invest. 1961 Mar;40:476–489. doi: 10.1172/JCI104275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HALL R. Stimulation of purine synthesis in vitro in the calf thyroid by thyroid-stimulating hormone. J Biol Chem. 1963 Jan;238:306–310. [PubMed] [Google Scholar]
  8. JUNGAS R. L., BALL E. G. Studies on the metabolism of adipose tissue; the stimulation of oxygen consumption by TSH preparations in relation to growth hormone and other pituitary fractions. Endocrinology. 1962 Jul;71:68–76. doi: 10.1210/endo-71-1-68. [DOI] [PubMed] [Google Scholar]
  9. KAPLAN M. E., COONS A. H., DEANE H. W. Localization of antigen in tissue cells; cellular distribution of pneumococcal polysaccharides types II and III in the mouse. J Exp Med. 1950 Jan 1;91(1):15-30, 4 pl. doi: 10.1084/jem.91.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LEVEY H. A., SOLOMON D. H. Studies on the metabolism of thyrotropin. Endocrinology. 1957 Jan;60(1):118–129. doi: 10.1210/endo-60-1-118. [DOI] [PubMed] [Google Scholar]
  11. LEZNOFF A., FISHMAN J., GOODFRIEND L., MCGARRY E., BECK J., ROSE B. Localization of flurescent antibodies to human growth hormone in human anterior pituitary glands. Proc Soc Exp Biol Med. 1960 Jun;104:232–235. doi: 10.3181/00379727-104-25790. [DOI] [PubMed] [Google Scholar]
  12. LEZNOFF A., FISHMAN J., TALBOT M., McGARRY E. E., BECK J. C., ROSE B. The cytological localization of ACTH in the human pituitary. J Clin Invest. 1962 Sep;41:1720–1724. doi: 10.1172/JCI104629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. MANCINI R. E., VILAR O., DELLACHA J. M., DAVIDSON O. W., CASTRO A. Histological localization in rat tissues of intravenously injected thyrotrophin labeled with a fluorescent dye. J Histochem Cytochem. 1961 May;9:271–277. doi: 10.1177/9.3.271. [DOI] [PubMed] [Google Scholar]
  14. MARSHALL J. M., Jr Localization of adrenocorticotropic hormone by histochemical and immunochemical methods. J Exp Med. 1951 Jul 1;94(1):21–30. doi: 10.1084/jem.94.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. OUCHTERLONY O. Antigen-antibody reactions in gels. Acta Pathol Microbiol Scand. 1949;26(4):507–515. doi: 10.1111/j.1699-0463.1949.tb00751.x. [DOI] [PubMed] [Google Scholar]
  16. RINDERKNECHT H. Ultra-rapid fluorescent labelling of proteins. Nature. 1962 Jan 13;193:167–168. doi: 10.1038/193167b0. [DOI] [PubMed] [Google Scholar]
  17. SALMON S., UTIGER R., PARKER M., REICHLIN S. The fate of I-131-labeled human growth hormone in the rabbit. Endocrinology. 1962 Apr;70:459–464. doi: 10.1210/endo-70-4-459. [DOI] [PubMed] [Google Scholar]
  18. SONENBERG M., KESTON A. S., MONEY W. L., RAWSON R. W. Radioactive thyrotropic hormone preparations. J Clin Endocrinol Metab. 1952 Oct;12(10):1269–1286. doi: 10.1210/jcem-12-10-1269. [DOI] [PubMed] [Google Scholar]
  19. SONENBERG M., KESTON A. S., MONEY W. L. Studies with labelled anterior pituitary preparations; adrenocorticotropin. Endocrinology. 1951 Feb;48(2):148–161. doi: 10.1210/endo-48-2-148. [DOI] [PubMed] [Google Scholar]
  20. WELLER T. H., COONS A. H. Fluorescent antibody studies with agents of varicella and herpes zoster propagated in vitro. Proc Soc Exp Biol Med. 1954 Aug-Sep;86(4):789–794. doi: 10.3181/00379727-86-21235. [DOI] [PubMed] [Google Scholar]

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