Hsc70-induced changes in clathrin-auxilin cage structure suggest a role for clathrin light chains in cage disassembly

The molecular chaperone, Hsc70, together with its cofactor, auxilin, facilitates the ATP-dependent removal of clathrin during clathrin-mediated endocytosis in cells. We have used cryo-electron microscopy to determine the 3D structure of a complex of clathrin, auxilin401-910 and Hsc70 at pH 6 in the presence of ATP, frozen within 20 seconds of adding Hsc70 in order to visualize events that follow the binding of Hsc70 to clathrin and auxilin before clathrin disassembly. In this map,we observe density beneath the vertex of the cage that we attribute to bound Hsc70. This density emerges asymmetrically from the clathrin vertex, suggesting preferential binding by Hsc70 for one of the three possible sites at the vertex. Statistical comparison with a map of whole auxilin and clathrin previously published by us reveals the location of statistically significant differences which implicate involvement of clathrin light chains in structural rearrangements which occur after Hsc70 is recruited. Clathrin disassembly assays using light scattering suggest that loss of clathrin light chains reduces the efficiency with which auxilin facilitates this reaction. These data support a regulatory role for clathrin light chains in clathrin disassembly in addition to their established role in regulating clathrin assembly. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Short- and long-term changes in corneal aberrations and axial length induced by orthokeratology in children are not correlated

PURPOSE: To assess the correlation between changes in corneal aberrations and the 2-year change in axial length in children fitted with orthokeratology (OK) contact lenses. METHODS: Thirty-one subjects 6 to 12 years of age and with myopia −0.75 to −4.00DS and astigmatism ≤1.00DC were fitted with OK. Measurements of axial length and corneal topography were taken at regular intervals over a 2-year period. Corneal topography at baseline and after 3 and 24 months of OK lens wear was used to derive higher-order corneal aberrations (HOA) that were correlated with OK-induced axial length changes at 2 years. RESULTS: Significant changes in C3, C4, C4, root mean square (RMS) secondary astigmatism and fourth and total HOA were found with both 3 and 24 months of OK lens wear in comparison with baseline (all P0.05). Coma angle of orientation changed significantly pre-OK in comparison with 3 and 24 months post-OK as well as secondary astigmatism angle of orientation pre-OK in comparison with 24 months post-OK (all P0.05). DISCUSSION: Short-term and long-term OK lens wear induces significant changes in corneal aberrations that are not significantly correlated with changes in axial elongation after 2-years.

Short-term and long-term changes in corneal power are not correlated with axial elongation of the eye induced by orthokeratology in children

PURPOSE: To assess the relationship between short-term and long-term changes in power at different corneal locations relative to the change in central corneal power and the 2-year change in axial elongation relative to baseline in children fitted with orthokeratology contact lenses (OK). METHODS: Thirty-one white European subjects 6 to 12 years of age and with myopia −0.75 to −4.00 DS and astigmatism ≤1.00 DC were fitted with OK. Differences in refractive power 3 and 24 months post-OK in comparison with baseline and relative to the change in central corneal power were determined from corneal topography data in eight different corneal regions (i.e., N[nasal]1, N2, T[temporal]1, T2, I[inferior]1, I2, S[superior]1, S2), and correlated with OK-induced axial length changes at two years relative to baseline. RESULTS: After 2 years of OK lens wear, axial length increased by 0.48±0.18 mm (P0.05). CONCLUSION: The reduction in central corneal power and relative increase in paracentral and pericentral power induced by OK over 2 years were not significantly correlated with concurrent changes in axial length of white European children.