Figure 4. Reversal of PA effects by expression of a type II PA phosphatase. (A and B) TEM analysis of photoreceptors overexpressing Pld alone (A) and Pld and LPP (B). Rhabdomeres (r) and membranous structures (v) are shown. Bars, 5 µm. (C) Western blots comparing the levels of Rh1 protein in the specified genotypes. Control bands show equivalent levels of protein loading. (D) Rh1 levels in cds1 mutants grown in 4 h light/20 h dark illumination cycles compared with cds1-coexpressing LPP. Samples from flies 0-, 2-, 4-, and 8-d-old are shown. Levels in wild-type flies are also shown. Levels of syntaxin are used as a loading control. (E and F) Immunofluorescence labeling of photoreceptors expressing wild-type (F) or K/R Pld (E). Rhabdomeres where the subcellular localization of Rh1 is normally restricted are shown in green in E; ER is labeled in red with an antibody to the KDEL epitope. In photoreceptors expressing Pld, Rh1 signals are no longer restricted to the rhabdomere but are distributed through the cell body. A critical time window for the effects of PLD overexpression. Bars, 10 μm. (G) Diagrammatic representation of the course of Drosophila pd. Time axis is shown as a percent of pd. In our conditions, this is 100 h. The temporal expression patterns Rh1-GAL4 and GMR-GAL4 are shown. The time of onset of phototransduction is also shown along with the duration of the rapid last phase of rhabdomere biogenesis. (H) Western blots from head extracts of control and experimental animals in which expression of Pld had been induced using heat shock GAL4 at the age specified (% pd). Animals not expressing Pld are marked as C, and those with Pld overexpression are marked as E. (top) Levels of Pld detected using a polyclonal antibody to this protein are shown. (bottom) Levels of Rh1 detected using a monoclonal antibody to this protein are shown. In the case of cds1, in which the elevation of PA is generated by sequential light-activated PLC and DGK, PA accumulation most likely starts at ∼70% pd when Rh1 and associated transduction components are first expressed, making the cell phototransduction competent. (H) Unit of measure, M r.
Figure 4.

Reversal of PA effects by expression of a type II PA phosphatase. (A and B) TEM analysis of photoreceptors overexpressing Pld alone (A) and Pld and LPP (B). Rhabdomeres (r) and membranous structures (v) are shown. Bars, 5 µm. (C) Western blots comparing the levels of Rh1 protein in the specified genotypes. Control bands show equivalent levels of protein loading. (D) Rh1 levels in cds1 mutants grown in 4 h light/20 h dark illumination cycles compared with cds1-coexpressing LPP. Samples from flies 0-, 2-, 4-, and 8-d-old are shown. Levels in wild-type flies are also shown. Levels of syntaxin are used as a loading control. (E and F) Immunofluorescence labeling of photoreceptors expressing wild-type (F) or K/R Pld (E). Rhabdomeres where the subcellular localization of Rh1 is normally restricted are shown in green in E; ER is labeled in red with an antibody to the KDEL epitope. In photoreceptors expressing Pld, Rh1 signals are no longer restricted to the rhabdomere but are distributed through the cell body. A critical time window for the effects of PLD overexpression. Bars, 10 μm. (G) Diagrammatic representation of the course of Drosophila pd. Time axis is shown as a percent of pd. In our conditions, this is 100 h. The temporal expression patterns Rh1-GAL4 and GMR-GAL4 are shown. The time of onset of phototransduction is also shown along with the duration of the rapid last phase of rhabdomere biogenesis. (H) Western blots from head extracts of control and experimental animals in which expression of Pld had been induced using heat shock GAL4 at the age specified (% pd). Animals not expressing Pld are marked as C, and those with Pld overexpression are marked as E. (top) Levels of Pld detected using a polyclonal antibody to this protein are shown. (bottom) Levels of Rh1 detected using a monoclonal antibody to this protein are shown. In the case of cds1, in which the elevation of PA is generated by sequential light-activated PLC and DGK, PA accumulation most likely starts at ∼70% pd when Rh1 and associated transduction components are first expressed, making the cell phototransduction competent. (H) Unit of measure, M r.

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