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Related Experiment Videos

Does thyrotropin cleave its cognate receptor?

Gregorio D Chazenbalk1, Chun-Rong Chen, Sandra M McLachlan

  • 1Autoimmune Disease Unit, Cedars-Sinai Research Institute and School of Medicine, University of California, Los Angeles, California 90048, USA.

Endocrinology
|September 10, 2003
PubMed
Summary
This summary is machine-generated.

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Thyroid-stimulating hormone (TSH) does not cleave the TSH receptor (TSHR) into subunits, contrary to recent findings. This study also highlights challenges in using flow cytometry to assess TSHR cleavage.

Area of Science:

  • Endocrinology
  • Molecular Biology
  • Cell Biology

Background:

  • A recent report suggested thyroid-stimulating hormone (TSH) cleaves the TSH receptor (TSHR) into subunits, challenging existing concepts.
  • This proposed cleavage has significant pathophysiological implications.

Purpose of the Study:

  • To re-evaluate the claim that TSH cleaves the TSH receptor (TSHR).
  • To assess the reliability of flow cytometry for quantifying TSHR cleavage.
  • To investigate the effects of TSH and a monoclonal antibody (MS-1) on TSHR subunit structure.

Main Methods:

  • Utilized CHO cell lines with high (TSHR-10,000) and conventional (TSHR-0) TSHR expression.
  • Performed flow cytometry with antibodies specific for single-chain TSHR and all TSHR forms after TSH or MS-1 pretreatment.

Related Experiment Videos

  • Employed covalent cross-linking of [125I]TSH to cell monolayers to directly determine TSHR subunit forms.

    • Main Results:

    • Flow cytometry did not show increased TSHR cleavage upon TSH pretreatment; instead, ligand occupancy (MS-1) and receptor downregulation were observed.
    • Covalent cross-linking confirmed that TSH had no effect on TSHR cleavage.
    • MS-1 slightly reduced spontaneous TSHR cleavage.

    Conclusions:

    • TSH does not alter the subunit structure of the TSH receptor.
    • The findings contradict a recent report on TSH-induced TSHR cleavage.
    • Difficulties and potential artifacts associated with the flow-cytometric assay for TSHR cleavage were identified.