Processing of the precursors of the human thyroid‐stimulating hormone receptor in various eukaryotic cells (human thyrocytes, transfected L cells and baculovirus‐infected insect cells)

Misrahi, Micheline; Ghinea, Nicolae; Sar, Sokhavuth; Saunier, Bertrand; Jolivet, André; Loosfelt, Hugues; Cérutti, Martine; Devauchelle, G.; Milgröm, Edwin

doi:10.1111/j.1432-1033.1994.tb18916.x

Cited by 110 publications

(149 citation statements)

References 40 publications

Supporting

Mentioning

136

Contrasting

Order By: Relevance

“…Establishment of MDCK Cell Lines Permanently Expressing FSH, LH, and TSH Receptors-MDCK cell lines were obtained and maintained as described (28,29) after cotransfection with expression vectors encoding either hFSHR (pSG5-hFSHR) (21), hTSHR (pSG5-hTSHR) (30), or pLHR (pCMV-LHRA) (8) and with the plasmid pSV-Neo which confers resistance to the antibiotic G418 (31). These clones were screened for receptor expression by immunocytochemistry using monoclonal anti-receptor antibodies.…”

Section: Methodsmentioning

confidence: 99%

“…Surface immunoprecipitation was performed as described (36) using monoclonal antibodies FSHR323 or LHR38 (30 g/ml) or polyclonal antibody TSHR 19-389 (dilution 1/300) added in the apical or in the basolateral compartment during the chase period. Extraction and purification of receptor-antibody complex were performed as described (30,36). All experiments were performed at least twice with triplicate samples.…”

Section: Verification Of the Polarized Phenotype Of Transfected Mdck mentioning

confidence: 99%

“…Purification and Western Blot Analysis of the Gonadotropin and TSH Receptors Expressed in MDCK Cells-These procedures were performed as described previously (20,21,26,30).…”

Section: Verification Of the Polarized Phenotype Of Transfected Mdck mentioning

confidence: 99%

“…Electrophoresis and Autoradiography-SDS-PAGE was performed as described (20,30). The gels were fixed and processed for fluorography.…”

Section: Verification Of the Polarized Phenotype Of Transfected Mdck mentioning

confidence: 99%

See 3 more Smart Citations

Basolateral Localization and Transcytosis of Gonadotropin and Thyrotropin Receptors Expressed in Madin-Darby Canine Kidney Cells

Beau

Misrahi

Gross

et al. 1997

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

؊ and cholera toxin. G s activation provoked a similar effect on LH receptor distribution in MDCK cells, whereas it did not modify the compartmentalization of the TSH receptor. Hormone-specific transcytosis was observed in MDCK cells expressing the gonadotropin (FSH and LH) receptors and was increased after cholera toxin administration.The gonadotropin (LH 1 and FSH) and thyrotropin (TSH) receptors belong to the large family of G-protein-coupled receptors (1-4). They possess the distinctive seven transmembrane spanning domains. However, they form a specific subgroup characterized by the presence of a large extracellular domain constituted by the repetition of leucine-rich motif (5). This ectodomain is responsible for the high affinity hormone binding (6 -9). Gonadotropin and TSH receptors are mainly coupled to G s and thus activate adenylate cyclase. The same receptors are also able to activate phospholipase C at high hormone concentrations. Mutations of the receptors have been described leading either to their constitutive activation (10 -12) or in contrast to a loss of receptor function (13)(14)(15).Little is known about the cellular trafficking of G-proteincoupled receptors in general and of this subgroup of receptors in particular. Ultrastructural immunocytochemistry has been used to analyze the internalization mechanisms of some receptors (16 -18). LH receptor-driven hormone transcytosis was observed through endothelial cells of testicular blood vessels (19). Recently immunocytochemistry using specific monoclonal antibodies demonstrated the basolateral distribution of the TSH receptor in thyroid cells (20) and of the FSH receptor in Sertoli cells (21), whereas the LH receptor was present all over the cell surface of thecal, granulosa, and luteal cells in the ovary and Leydig cells in the testes (22,23).The question was thus raised as to whether these differences in cellular distribution were due to differences in the structure of receptors or simply secondary to the fact that thyroid follicular cells and Sertoli cells are polarized, whereas the LH receptor-expressing cells are not polarized. Another question raised was whether the mechanisms of receptor basolateral delivery were cell-specific or whether the receptors contained specific signals that could direct them to this membrane compartment in any polarized cell. To answer these questions we have established MDCK cell lines that express the FSH, LH, and TSH receptors. Using these models we have analyzed the distribution of the receptors in polarized monolayers. MATERIALS AND METHODSCell Culture-MDCK cells (type II) were seeded and grown on coverslips (Nunc) or filters (0.4-m polycarbonate, tissue culture-treated, Transwell costar) as described previously (24,25).Antireceptor Antibodies-Mouse monoclonal antibodies FSHR323 (21), LHR38 (26),27) recognize an epitope in the extracellular domain of the FSH, LH, and TSH receptors, respectively. Antibody T3-365 has been raised against the intracellular domain of the TSH receptor (20). The rabbit polyclonal TSHR...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Verification Of the Polarized Phenotype Of Transfected Mdck mentioning

confidence: 99%

“…Purification and Western Blot Analysis of the Gonadotropin and TSH Receptors Expressed in MDCK Cells-These procedures were performed as described previously (20,21,26,30).…”

Section: Verification Of the Polarized Phenotype Of Transfected Mdck mentioning

confidence: 99%

“…Electrophoresis and Autoradiography-SDS-PAGE was performed as described (20,30). The gels were fixed and processed for fluorography.…”

Section: Verification Of the Polarized Phenotype Of Transfected Mdck mentioning

confidence: 99%

See 2 more Smart Citations

Basolateral Localization and Transcytosis of Gonadotropin and Thyrotropin Receptors Expressed in Madin-Darby Canine Kidney Cells

Beau

Misrahi

Gross

et al. 1997

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…1). The 65-to 75-kDa band occasionally seen may reflect a subunit of the receptor protein (26,27) or possibly nonglycosylated TSH receptor from cell lysates. As shown in Fig.…”

Section: Immunoprecipitation Of Tshr From Intestinal Lymphoid Cells mentioning

confidence: 99%

The Thyrotropin (Thyroid-Stimulating Hormone) Receptor Is Expressed on Murine Dendritic Cells and on a Subset of CD45RBhigh Lymph Node T Cells: Functional Role for Thyroid-Stimulating Hormone During Immune Activation

Bagriacik

Klein

2000

The Journal of Immunology

View full text Add to dashboard Cite

Thyroid-stimulating hormone (TSH), a central neuroendocrine mediator of the hypothalamus-pituitary-thyroid axis, has been shown to affect various aspects of immunological development and function. To gain a better understanding of TSH involvement within the mammalian immune system, the expression and distribution of the TSH receptor (TSHr) has been studied by immunoprecipitation and by flow cytometric analyses. Using highly enriched populations of B cells, T cells, and dendritic cells, trace amounts of TSHr were precipitated from B cells and T cells, whereas high levels of TSHr were precipitated from the dendritic cell fraction. Flow cytometric analyses of TSHr expression on splenic and lymph node T cells revealed a major difference between those tissues in that only 2–3% of splenic T cells were TSHr+, whereas 10–20% of CD4+8− and CD4−8+ lymph node T cells expressed the TSHr, which was exclusively associated with CD45RBhigh cells and was not expressed during or after activation. The TSHr was not present on cells of the immune system during fetal or neonatal life. However, recombinant TSHβ was found to significantly enhance the phagocytic activity of dendritic cells from adult animals and to selectively augment IL-1β and IL-12 cytokine responses of dendritic cells following phagocytic activation. These findings identify a novel immune-endocrine bridge associated with professional APCs and naive T cells.

show abstract

The Expression of Soluble, Full-Length, Recombinant Human TSH Receptor in a Prokaryotic System

Busuttil

Turney²,

Frauman

2001

Protein Expression and Purification

View full text Add to dashboard Cite

Processing of the precursors of the human thyroid‐stimulating hormone receptor in various eukaryotic cells (human thyrocytes, transfected L cells and baculovirus‐infected insect cells)

Cited by 110 publications

References 40 publications

Basolateral Localization and Transcytosis of Gonadotropin and Thyrotropin Receptors Expressed in Madin-Darby Canine Kidney Cells

Basolateral Localization and Transcytosis of Gonadotropin and Thyrotropin Receptors Expressed in Madin-Darby Canine Kidney Cells

The Thyrotropin (Thyroid-Stimulating Hormone) Receptor Is Expressed on Murine Dendritic Cells and on a Subset of CD45RBhigh Lymph Node T Cells: Functional Role for Thyroid-Stimulating Hormone During Immune Activation

The Expression of Soluble, Full-Length, Recombinant Human TSH Receptor in a Prokaryotic System

Contact Info

Product

Resources

About