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DOI: 10.1055/s-0034-1372102
Phenotyping the mouse thyroid gland by semi-quantitative cell biology
Introduction: The thyroid gland is the thyroid hormone producing organ of mammals and it consists of thyroid follicles which, together with blood vessels, represent the functional units. Phenotypic analysis of thyroid follicle cells can shed light on the functional state of the thyroid gland and will therefore contribute to the basic understanding of this important endocrine tissue.
Methods: In this study we investigated thyroid glands of mice deficient in thyroid hormone transporters, such as Lat2, Mct8 and/or Mct10, and compared those to mice with impaired thyroglobulin processing due to cathepsin-deficiencies, while normalizing by the use of wild type littermates. Thus, we established a platform to analyse (immuno-) labelled cryo-sections prepared from thyroid glands of mice with different genotypes using automated, quantitative imaging software. The read-out parameters relevant for phenotyping comprised thyroid follicle diameters, cell numbers per follicle, and extensions of the epithelial monolayers, while the supporting blood circulation was approached by analysing endothelial cells. As molecular markers of the thyroid functional state, the TSH receptor, the sodium iodide symporter NIS, thyroglobulin-processing proteases, TH transporters, collagen IV of the basal lamina, and endothelial CD-31 were investigated.
Results: Our data indicates that thyroids lacking thyroglobulin-processing enzymes or TH transporters can be described in a qualitative and semi-quantitative manner by this phenotyping method which detects changes between different genotypes significantly.
Conclusion: Hence, the imaging and analysis pipeline established herein can be used as a powerful tool for providing data on the functional state of the thyroid, thus generating insights into thyroid cell biology with a rapid and versatile method.