Direct measurement of diffusion in olfactory cilia using a modified FRAP approach.
The diffusion coefficient of fluorescein in detached cilia of Xenopus laevis olfactory receptor neurons was measured using spatially-resolved FRAP, where the dye along half of the ciliary length was photobleached and its spatiotemporal fluorescence redistribution recorded. Fitting a one-dimensional numerical simulation of diffusion and photobleaching for 35 cilia resulted in a mean value of the diffusion coefficient (1.20 ± 0.23) · 10(-10)m(2)/s and thus a reduction by a factor of 3.4 compared to free diffusion in aqueous solution.
Department of Neurophysiology and Cellular Biophysics, University of Göttingen, Göttingen, Germany. malevra@gwdg.de
Mihai
M
Alevra
Peter
P
Schwartz
Detlev
D
Schild
D000818:Animals; D002923:Cilia / Q000378:metabolism* / Q000648:ultrastructure; D004058:Diffusion; D019793:Fluorescein; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods*; D005456:Fluorescent Dyes; D007700:Kinetics; D007814:Larva / Q000033:anatomy & histology / Q000378:metabolism*; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D018034:Olfactory Receptor Neurons / Q000378:metabolism* / Q000648:ultrastructure; D038761:Photobleaching; D012996:Solutions; D046509:Tissue Culture Techniques; D014867:Water; D014982:Xenopus laevis / Q000033:anatomy & histology / Q000378:metabolism*
D005456:Fluorescent Dyes; D012996:Solutions; D014867:Water; D019793:Fluorescein