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. 2014 Apr 29;12(4):e1001847. doi: 10.1371/journal.pbio.1001847

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© 2014 Wang et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Schematic showing that mutations affecting PLC (norpA) or the μ3 subunit of the AP-3 complex (cm) lead to PR degeneration due to Rh1 accumulation in late endosomes. (B) Immunostaining for F-actin to mark rhabdomeres in the retina of control (w¹¹¹⁸, left panel), norpA^P24 mutants (middle left panel), norpA^P24;actin-GAL>UAS-Vps35,UAS-w RNAi (middle right panel), or norpA^P24;actin-GAL>UAS-Vps26,UAS-w RNAi (right panel) flies upon 6 d of continuous exposure to whole-spectrum light. The number of rhabdomeres and organization of ommatidia of norpA^P24 mutants are severely reduced or impaired upon a 6-d exposure to light (middle left panel) but remain largely intact when Vps35 (middle right panel) or Vps26 (right panel) is overexpressed. OE, overexpression. Scale bar, 5 µm. (C) Quantification of the rhabdomere number per ommatidium for the indicated genotypes in (B). Sixty ommatidia from six animals were examined for each genotype. Student's t test; error bars represent SEM; ** p<0.01. (D) Immunostaining for F-actin to mark rhabdomeres in the retina of control flies overexpressing lacZ (actin-GAL4>UAS-lacZ, top left panel), Vps35 (actin-GAL4>UAS-Vps35, top middle panel), or Vps26 (actin-GAL4>UAS-Vps26, top right panel), or cm¹ mutants overexpressing lacZ (cm¹;actin-GAL4>UAS-lacZ, bottom left panel), Vps35 (cm¹;actin-GAL4>UAS-Vps35, bottom middle panel), or Vps26 (cm¹;actin-GAL4>UAS-Vps26, bottom right panel) upon constant white light exposure for 6 d. The UAS-w RNAi is co-expressed in all animals. The organization and size of cm¹ mutant rhabdomeres are strongly impaired upon light exposure. Vps35 or Vps26 OE suppresses the PR morphological defects in cm¹ mutants. Scale bar, 5 µm. (E) Quantification of the rhabdomere number per ommatidium in the indicated genotypes. Sixty ommatidia from six animals were examined for each genotype. Student's t test; error bars represent SEM; ** p<0.01; ns, no significance. (F) ERG traces of control overexpressing lacZ (actin-GAL4>UAS-lacZ, top left panel), Vps35 (actin-GAL4>UAS-Vps35, top middle panel), or Vps26 (actin-GAL4>UAS-Vps26, top right panel), or cm¹ mutants overexpressing lacZ (cm¹;actin-GAL4>UAS-lacZ, bottom left panel), Vps35 (cm¹;actin-GAL4>UAS-Vps35, bottom middle panel), or Vps26 (cm¹;actin-GAL4>UAS-Vps26, bottom right panel) flies. The w RNAi is co-expressed in each animal. After light exposure for 6 d, flies were transferred to the dark for 1 d to avoid visual adaptation to light. cm¹ mutants show a reduction of ERG amplitudes and loss of the on- and off-transients. Vps35 or Vps26 OE partially restores the on and offs and ERG amplitudes. (G) Quantification of ERG amplitudes of the flies indicated in (F). Ten ERG traces were recorded for each genotype. ERG amplitudes are normalized to control PRs. Student's t test; error bars represent SEM; * p<0.05; ns, no significance.