Effect of thyrotrophin on epidermal growth factor receptors in monolayer cultures of porcine thyroid cells.
Primary cultures of porcine thyroid cells, grown as monolayers, showed saturable binding of epidermal growth factor (EGF). Scatchard analysis resolved the binding to a high-affinity/low-capacity site (dissociation constant = 0.17 nmol/l, maximal binding capacity = 1.67 pmol/10(6) cells) and a low-affinity/high-capacity site. Preincubation of thyroid monolayers with TSH for 3 days caused an increase in binding of 125I-labelled EGF due to an increase in the number of receptors, with the binding affinity unchanged. This effect was dose-dependent within the range of TSH concentrations 0.01-100 mu/l. The same effect was seen with dibutyryl cyclic AMP (10-1000 mumol/l). When the protein synthesis inhibitor cycloheximide was included in the TSH preincubation, the increase in EGF binding was abolished. The TSH effect on EGF binding was not mediated by thyroid hormones, since neither thyroxine (T4) nor tri-iodothyronine (T3) at 0.1 nmol/l-10 mumol/l could mimic the effect of TSH, nor could antisera to T3 or T4 neutralize the effect of TSH. The concentration of extracellular iodide (10 nmol/l-10 mmol/l) had no effect on the binding of 125I-labelled EGF. The results demonstrate that TSH increases the number of receptor sites for binding of EGF to thyroid monolayers in vitro. This effect is dependent upon protein synthesis and is mediated by cyclic AMP but not by thyroid hormones or iodide. This effect on binding of EGF may contribute to the trophic action of TSH.
D000818:Animals; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000187:drug effects*; D013961:Thyroid Gland / Q000187:drug effects* / Q000378:metabolism; D013972:Thyrotropin / Q000494:pharmacology*; D013974:Thyroxine / Q000494:pharmacology; D014284:Triiodothyronine / Q000494:pharmacology
D014284:Triiodothyronine; D004815:Epidermal Growth Factor; D013972:Thyrotropin; D003513:Cycloheximide; D066246:ErbB Receptors; D013974:Thyroxine