Table 1. Individual GPCR and cilia parameters: low frame rate (3–5 fps) tracking.
Ciliuma | Tracking time | Cilium length | GPCR densityb | GPCRs on ciliumc | Amd | Daxe | Dmf | MIg | fcsh | MSDinti |
---|---|---|---|---|---|---|---|---|---|---|
min | μm | μm−2 | % | μm2/s | μm2/s | μm2 | ||||
R1 | 10.5 | 3.7 | 269.2 | 932 | 0.24 | 0.074 | 0.148 | 0.77 | 1.0 | 0.079 |
R2 | 3.0 | 4.2 | 1,487.5 | 5,833 | 1.34 | 0.026 | 0.051 | 0.35 | 0.7 | 0.020 |
R3 | 4.0 | 1.9 | 4,965.2 | 8,735 | 4.47 | 0.035 | 0.069 | 0.78 | 1.0 | 0.020 |
R4 | 17.1 | 3.7 | 2,790.7 | 9,721 | 2.51 | 0.135 | 0.270 | 0.50 | 1.0 | 0.079 |
R5 (1) | 12.0 | 2.9 | 1,653.2 | 4,592 | 1.49 | 0.015 | 0.030 | 0.61 | 1.0 | 0.009 |
R5 (2) | 12.0 | 0.081 | 0.163 | 0.34 | 1.0 | 0.049 | ||||
R6 | 8.4 | 3.8 | 1,607.6 | 5,795 | 1.45 | 0.030 | 0.059 | 0.42 | 0.9 | 0.018 |
R7 | 4.5 | 4.3 | 30.5 | 123 | 0.03 | 0.145 | 0.290 | 0.26 | 1.0 | 0.070 |
R8 | 9.7 | 2.4 | 873.2 | 1,969 | 0.79 | 0.095 | 0.189 | 0.70 | 1.0 | 0.056 |
Mean R | 3.6 | 1,744.9 | 4,713 | 1.54 | 0.071 | 0.141 | 0.53 | 0.96 | 0.045 | |
SEM R | 0.3 | 512.5 | 1,172 | 0.47 | 0.016 | 0.032 | 0.07 | 0.03 | 0.009 | |
S1 | 11.9 | 5.6 | 557.5 | 2,947 | 0.50 | 0.085 | 0.170 | 0.39 | 1.0 | 0.027 |
S2 (1) | 12.7 | 4.8 | 10,676.0 | 48,368 | 9.61 | 0.167 | 0.334 | 0.62 | 1.0 | 0.099 |
S2 (2) | 12.7 | 0.180 | 0.361 | 0.55 | 1.0 | 0.099 | ||||
S3 (1) | 9.2 | 7.4 | 6,534.6 | 45,267 | 5.88 | 0.063 | 0.126 | 0.63 | 1.0 | 0.037 |
S3 (2) | 9.2 | 0.032 | 0.065 | 0.62 | 1.0 | 0.020 | ||||
S4 | 28.3 | 7.0 | 7,292.1 | 47,937 | 6.56 | 0.079 | 0.157 | 0.21 | 0.7 | 0.047 |
S5 | 14.7 | 8.1 | 1,591.5 | 12,077 | 1.43 | 0.141 | 0.282 | 0.28 | 1.0 | 0.079 |
S6 | 7.5 | 10.0 | 1,761.3 | 16,631 | 1.59 | 0.075 | 0.149 | 0.50 | 0.8 | 0.064 |
S7 | 14.8 | 4.7 | 2,591.2 | 11,404 | 2.33 | 0.109 | 0.217 | 0.44 | 1.0 | 0.075 |
S8 | 4.1 | 8.2 | 2,612.2 | 20,191 | 2.35 | 0.121 | 0.242 | 0.39 | 0.9 | 0.096 |
S9 | 5.2 | 6.4 | 1,461.0 | 8,748 | 1.31 | 0.086 | 0.172 | 0.19 | 0.3 | 0.010 |
S10 | 6.3 | 7.2 | 2,412.5 | 16,257 | 2.17 | 0.138 | 0.276 | 0.49 | 1.0 | 0.099 |
S11 (1) | 16.0 | 7.3 | 1,487.1 | 10,205 | 1.34 | 0.129 | 0.258 | 0.46 | 1.0 | 0.056 |
S11 (2) | 16.0 | 0.111 | 0.222 | 0.56 | 1.0 | 0.056 | ||||
Mean S | 7.2 | 3,543.4 | 21,821 | 3.19 | 0.108 | 0.216 | 0.45 | 0.91 | 0.062 | |
SEM S | 0.5 | 1,061.4 | 5,640 | 0.86 | 0.011 | 0.022 | 0.04 | 0.05 | 0.008 |
Individual cilia: R, Rhoi3S; S, SSTR3. Numbers in parentheses identify individual Qdot-GPCR for cilia in which more than one was labeled.
GPCR density was estimated by calibration of the optical system with recombinant EGFP of known concentrations. Confinement of the EGFP label to the ciliary membrane and the sampling by the optical system were accounted for as described previously (Peet et al., 2004).
Total number of GPCRs within the given cilium.
Membrane area fraction occupied by the expressed GPCRs.
Dax is the 1D axial diffusion coefficient estimated from the initial slope of the MSD(τ) plot for an individual Qdot-GPCR.
Effective membrane diffusion coefficient: Dm ≈ 2Dax. Dax underestimates the true 2D membrane diffusion coefficient approximately twofold. Note the similarity of the mean microscopic Rhoi3S diffusion coefficient in ciliary membranes, Dm, reported here to the mean macroscopic lateral diffusion coefficient estimated for rhodopsin within discrete lobules of photoreceptor disc membranes, Dlat = 0.130 ± 0.012 µm2/s (Najafi et al., 2012a).
Mobility index (see text).
Fraction of cilium length sampled by the indicated GPCR.
MSD value at the y-intercept of the linear regression of the first two MSD(τ) points.