title: TSH induces co-localization of TSH receptor and Na/K-ATPase in human erythrocytes.
abstract: Thyroid stimulating hormone (TSH) binds to a specific TSH receptor (TSHR) which activates adenylate cyclase and increases cAMP levels in thyroidal cells. Recent studies have reported the presence of TSH receptor in several extra-thyroidal cell types, including erythrocytes. We have previously suggested that TSH is able to influence the erythrocyte Na/K-ATPase ouabain binding properties through a receptor mediated mechanism. The direct interaction of TSH receptor with the Na/K-pump and a functional role of TSHR in erythrocytes was not demonstrated. The interaction of TSH receptor with Na/K-pump and a TSHR functional role are not yet demonstrated in erythrocytes. In this study, we examined the interaction between the two receptors after TSH treatment using immunofluorescence coupled to confocal microscopy and a co-immunoprecipitation technique. The cAMP dependent signalling after TSH treatment was measured to verify TSHR functionality. We found that TSH receptor and Na/K-ATPase are localized on the membranes of both erythrocytes and erythrocyte ghosts; TSH receptor responds to TSH treatment by increasing intracellular cAMP levels from two to tenfold. In ghost membranes TSH treatment enhances up to three fold co-localization of TSHR with Na/K-ATPase and co-immunoprecipitation confirms their direct physical interaction. In conclusion our results are compatible with the existence, in erythrocytes, of a functional TSHR that interacts with Na/K-ATPase after TSH treatment, thus suggesting a novel cell signalling pathway, potentially active in local circulatory control.
authors.affiliation: Institute of Clinical Physiology, CNR, Pisa, Italy. balzan@ifc.cnr.it
authors.forename: Silvana
authors.initials: S
authors.lastname: Balzan
authors.forename: Renata
authors.initials: R
authors.lastname: Del Carratore
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Nicolini
authors.forename: Francesca
authors.initials: F
authors.lastname: Forini
authors.forename: Valter
authors.initials: V
authors.lastname: Lubrano
authors.forename: Marcella
authors.initials: M
authors.lastname: Simili
authors.forename: Pier Alberto
authors.initials: PA
authors.lastname: Benedetti
authors.forename: Giorgio
authors.initials: G
authors.lastname: Iervasi
mesh_terms: D000242:Cyclic AMP / Q000378:metabolism; D004912:Erythrocytes / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D011989:Receptors, Thyrotropin / Q000032:analysis* / Q000378:metabolism / Q000502:physiology; D015398:Signal Transduction; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis* / Q000378:metabolism / Q000502:physiology; D013972:Thyrotropin / Q000494:pharmacology*
chemical_list: D011989:Receptors, Thyrotropin; D013972:Thyrotropin; D000242:Cyclic AMP; D000254:Sodium-Potassium-Exchanging ATPase