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Acta Physiologica 2010; Volume 200, Supplement 681
Abstracts of the 61st National Congress of the Italian Physiological Society
9/15/2010-9/17/2010
Varese, Italy
ROLE OF ADDUCIN IN THE EXPRESSION OF CFTR IN NA+-SENSITIVE HYPERTENSION
Abstract number: P32
SASSONE1 F, GARAVAGLIA1 ML, MONDINI1 A, BAZZINI1 C, RODIGHIERO2 S, VEZZOLI1 V, MEYER1 G
1Dept of Biomolecular Sciences and Biotechnology, Univ. of Milan, Italy
2Fondazione Filarete, Milan, Italy
Alterations of NaCl absorption in the kidney lead to the development of Na+-sensitive hypertension. Milan Hypertensive Rats (MHS), an animal model for this pathology, are characterized by a raise of Na+,K+ ATPase activity and expression, due to a mutation of the cytoskeletal protein adducin, whose mutations cause hypertension in humans too. Patch-clamp experiments on MHS and MNS DCT cells revealed the activity of a Cl- channel with the same characteristics of CFTR, whose density and activity were significantly increased in MHS rats. As CFTR is influenced by cytoskeletal proteins, we tested the effect on CFTR of wild-type (WT) and hypertension-related G460W adducin variant. By means of whole-cell patch-clamp experiments, performed on NU12 (HEK cells stably transfected with the human WT adducin) and HU33 (HEK cells stably transfected with the G460W mutation) cells, a significantly higher and faster CFTR Cl- current was observed. Western blot and immunofluorescence experiments revealed that CFTR was more expressed in the plasmamembrane in HU than NU, confirmed by cell-attached experiments. By means of FRET and coimmunoprecipitation experiments a weak interaction between CFTR and adducin was demonstrated in HEK cells. As CFTR undergoes trafficking from and to the plasma membrane, FRAP experiments were performed to study whether the mobility of CFTR is different in presence of WT or G460W adducin. As a conclusion adducin expression influences CFTR expression and activity.
To cite this abstract, please use the following information:
Acta Physiologica 2010; Volume 200, Supplement 681 :P32