Mapping mechanotransduction mediated by primary cilia at nanoscale (360G-Wellcome-109724_Z_15_Z)

Mechanical stimulation is one of the key physiological stimuli. Abnormalities in mechanical load or in cellular mechano-sensitivity have been linked to diseases such as osteoporosis, osteoarthritis or polycystic kidney disease. No effective treatment exists for these conditions as our understanding of mechanotransduction is still incomplete. A unique membrane protrusion called primary cilium, and a number of membrane receptors sensitive to mechanical and chemical stimuli have been already implic ated in the mechanotransduction, however the signalling pathways remain uncertain. We hypothesize that the actual role of receptors involved in mechanosensing is affected by their precise location within the fine ultrastructure of the primary cilium. To investigate this hypothesis we propose the use of scanning nanopipette microscopy capable of functional assessment of membrane channel and receptor activity with nanoscale spatial resolution. We will establish recordings of channel activity in re sponse to mechanical stimulation at precisely defined locations on primary cilium and develop technique for mapping local sensitivity to chemical stimuli by means of highly localised dosing via the nanopipette. The established techniques will be subsequently used to gather preliminary data on the distribution of mechano-chemical receptors to support research proposal aimed at understanding the reorganisation of mechanotransduction in ciliated cells in disease.