Overexpression of functional TrkA receptors after internalisation in human airway smooth muscle cells.
Trafficking of the TrkA receptor after stimulation by NGF is of emerging importance in structural cells in the context of airway inflammatory diseases. We have recently reported the expression of functional TrkA receptors in human airway smooth muscle cells (HASMC). We have here studied the TrkA trafficking mechanisms in these cells. TrkA disappearance from the cell membrane was induced within 5 min of NGF (3pM) stimulation. Co-immunoprecipitation of clathrin-TrkA was revealed, and TrkA internalisation inhibited either by clathrin inhibitors or by siRNA inducing downregulation of endogenous clathrin. TrkA internalised receptors were totally degraded in lysosomes, with no recycling phenomenon. Newly synthesized TrkA receptors were thereafter re-expressed at the cell membrane within 10 h. TrkA re-synthesis was inhibited by blockade of clathrin-dependent internalisation, but not of TrkA receptors lysosomal degradation. Finally, we observed that NGF multiple stimulations progressively increased TrkA expression in HASMC, which was associated with an increase in NGF/TrkA-dependent proliferation. In conclusion, we show here the occurrence of clathrin-dependent TrkA internalisation and lysosomal degradation in the airway smooth muscle, followed by upregulated re-synthesis of functional TrkA receptors and increased proliferative effect in the human airway smooth muscle. This may have pathophysiological consequences in airway inflammatory diseases.
Freund-Michel
Véronique
V
EA 3771 "Inflammation and environment in asthma", University Louis Pasteur-Strasbourg-I, Faculty of Pharmacy, Illkirch, France. veronique.michel@u-bordeaux2.fr
Frossard
Nelly
N
D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D020932:Nerve Growth Factor / Q000494:pharmacology; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics; D020917:Receptor, trkA / Q000235:genetics / Q000378:metabolism*
D002966:Clathrin; D012333:RNA, Messenger; D020932:Nerve Growth Factor; D020917:Receptor, trkA