Sulfation affects apical extracellular matrix organization during development of the Drosophila embryonic salivary gland tube
- Department of Biological Sciences, Louisiana State University, United States
- Shared Instrumentation Facility, Louisiana State University, United States
Figures
Figure 1 with 4 supplements
PAPS synthetase (Papss) mutants show defects in salivary gland (SG) lumen expansion.
(A) Stage 16 SGs immunostained for Stranded at Second (SAS). (B) In situ hybridization using an antisense probe for Papss mRNA in wild-type (WT) embryos. (C–H) Stage 14 and stage 16 SGs immunostained for Ecad and CrebA. Yellow asterisks indicate regions of irregular, non-uniform expansion (D, E, H) or over-expansion (G) of the SG lumen. (I, I’) Quantification of different SG lumen phenotypes. (J) Quantification of SG lumen length. n=7–11 SGs for each genotype. See the Materials and methods for the sample size of each genotype. One-way ANOVA test with multiple comparisons (*p<0.05; ns, non-significant). (K, K’) Quantification of the SG lumen diameter (K) and the coefficient of variation for each SG (K’). The diameter is measured in three different regions of the lumen. The average diameter, as well as the individual data points, are shown. The same samples are used for the quantifications in J-K’. One-way ANOVA test with multiple comparison (***p<0.001; ns, non-significant) (L) Quantification of SG lumen area. n=7–16 SGs for each genotype. One-way ANOVA test with multiple comparison (***p<0.001; ****p<0.0001; ns, non-significant). Horizontal lines indicate mean values.
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Figure 1—source data 1
Raw data for the quantification of salivary gland (SG) lumen length, lumen diameter, lumen area, and coefficient variations for lumen diameter.
See Figure 1J-L for the graphs.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig1-data1-v1.xlsx
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Figure 1—source data 2
Raw data for the quantification of salivary gland (SG) lumen length, lumen diameter, lumen area, and coefficient variations for lumen diameter.
See Figure 1J-L for the graphs.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig1-data2-v1.xlsx
Figure 1—figure supplement 1
PAPS synthetase (Papss) mutants encode a stop codon in the ATP sulfurylase domain.
(A) Cartoon of the Papss-PA isoform showing the adenosine 5’-phosphosulfate (APS) kinase and ATP sulfurylase domains. A red asterisk indicates the site of the W271->stop mutation. (B) Amino acid sequences for WT and Papss2 sequences for the Papss-PA isoform. Green text, residues of the APS kinase domain. Blue text, residues of the ATP sulfurase domain. Red asterisks, stop codon. Note that Papss2 also has a secondary mutation (G->A) at G79 of the transcript, resulting in a silent mutation.
Figure 1—figure supplement 2
Sulfotyrosine signals are reduced in PAPS synthetase (Papss) mutants.
(A, B) Confocal images of salivary glands (SGs) immunostained for sulfotyrosine and Ecad in stage 16 wild-type (WT) (A) and Papss mutant embryos. Magnified images are shown for boxed regions. Yellow arrowheads indicate sulfotyrosine-positive cytoplasmic puncta in WT SG cells. Cyan arrowheads, sulfotyrosine at the apical membrane. Asterisks, almost no sulfotyrosine puncta are present in the cytoplasm in Papss mutants. (C) Confocal images of stage 16 SGs immunostained for GFP and sulfotyrosine. Arrows, ManII-GFP, and sulfotyrosine puncta co-localize.
Figure 1—figure supplement 3
Some PAPS synthetase (Papss) mutant embryos show defective salivary gland (SG) formation and whole embryonic defects.
(A–D) Confocal images of stages 11–16 embryos immunostained using CrebA. (A) wild-type (WT). (B) Papss, normal embryonic morphology. (C) Papss/Df, normal embryonic morphology. (D) Papss, severely defective whole embryo morphology. (E, F) Quantification of the percentage of embryos with whole embryonic defects. (E) Quantification of combined stages 11–16. (F) Quantification of individual stages for stages 14–16. (G–H) Quantification of the percentage of embryos with SG invagination defects/mispositioned SG. (G) Quantification of combined stages 12–16. (H) Quantification of individual stages for stages 14–16.
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Figure 1—figure supplement 3—source data 1
Raw data for quantifying defects in the whole embryonic morphology and salivary gland (SG) positioning in wild-type (WT), Papss homozygous, transheterozygous over a deficiency, and heterozygous mutant embryos.
See Figure 1—figure supplement 3 for graphs.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig1-figsupp3-data1-v1.xlsx
Figure 1—figure supplement 4
Several splice forms of PAPS synthetase (Papss) are expressed in the salivary gland (SG).
(A) Eight annotated splice forms of Papss are reported by FlyBase. The blue bars indicate the regions used to make anti-sense probes that recognize different subsets of splice forms. Green and gray boxes indicate coding and non-coding exons, respectively. Thin lines indicate introns. (B) Fluorescence in situ hybridization with probes 1–4 (cyan). SGs are marked with CrebA (green) and Crb (magenta). Higher magnification of the mRNA signals within the SG is shown in the insets. Yellow dotted lines indicate the SG boundary.
Figure 2 with 2 supplements
Mutations in PAPS synthetase (Papss) result in mislocalized Crumbs (Crb) and disruption of the apical extracellular matrix (aECM) architecture.
(A) Cartoon showing the subcellular localization of Crb in salivary gland (SG) cells. (B, C) Stage 16 SGs immunostained for Crb and E-cadherin (Ecad). Magnified images are shown for the yellow dotted boxed regions. (D) Quantification of the Crb signal intensity ratio between the subapical region (SAR) and apical-medial regions of SG cells. (n=7, WT; n=6, Papss2 (4–8 cells per SG)). Student’s t-test with Welch’s correction (***p<0.001). (E–G) Stage 16 SGs stained for Ecad and wheat germ agglutinin (WGA). Magnified images are shown for the boxed regions. Arrows, WGA localization at the apical membrane in WT (yellow) and Papss mutant (cyan) SGs. Yellow arrowhead in E, WGA localizes as thin filaments in the lumen of WT SG. Cyan arrowheads, WGA localizes as a highly condensed structure in the thin luminal region of the Papss SG. White arrowheads, WGA localizes as a condensed, filamentous structure in the expanded luminal region of the Papss SG. Yellow arrowheads in G, WGA localizes as a mildly condensed filament in the lumen of sage >Papss PE; Papss2 SG. (H, I) Transmission electron microscopy (TEM) images of stage 15 SGs. SGs are pseudo-colored in magenta. Magnified images of the luminal areas (boxed regions) are shown at the bottom. Yellow asterisk in H, electron-dense luminal material in WT SG. Cyan asterisks in I, condensed electron-dense luminal material in the Papss mutant SG. Green asterisk, a large luminal area lacking electron-dense material in the Papss mutant SG.
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Figure 2—source data 1
Raw data for the quantification of the mean gray values of Crumbs (Crb) levels and the ratio of the values between the subapical and apical-medial regions.
See Figure 2D for the graph.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Adherens junctions (AJs) and septate junctions (SJs) are intact in PAPS synthetase (Papss) mutants.
(A) Cartoon showing the subapical and junctional regions in the PAPS synthetase (SG) epithelial cell from the lateral view and localization of the key protein markers Crb, E-cadherin (Ecad), and discs large (Dlg). (B, C) Confocal images of stage 16 SGs immunostained for Ecad. Magnified images are shown for boxed regions. Yellow asterisks indicate the localization of Ecad to AJs in both wild-type (WT) (B) and Papss mutant (C) SGs. (D, E) Confocal images of stage 16 SGs immunostained for Dlg. Images show the luminal surface view. Cyan asterisks indicate the localization of Dlg at SJs in both WT (D) and Papss mutant (E) SGs. SJs in Papss mutants occasionally appear slightly longer in the expanded luminal region compared to WT or the thin luminal region of Papss mutants, possibly due to stretching of cells with lumen expansion.
Figure 2—figure supplement 2
Original, raw transmission electron microscopy (TEM) images of wild-type (WT) and Papss mutant salivary glands (SGs).
Figure 3 with 3 supplements
Loss of PAPS synthetase (Papss) results in disorganized Golgi structures and defects in intracellular trafficking components.
(A–D) Stage 16 salivary glands (SGs) stained with wheat germ agglutini (WGA). Magnified images are shown for the boxed regions. Yellow arrows, WGA signals as cytoplasmic puncta. Cyan arrows, WGA signals in the nuclear envelope. (E–H) Three-dimensional (3D) reconstruction of cytoplasmic WGA puncta in stage 16 SGs. (I, I’) Quantification of the number of WGA puncta (I) and signal intensity (I’) within SG cells. n=5 SGs for all genotypes, except WT in G (n=6). One-way ANOVA test with multiple comparisons (**p<0.01; ***p<0.001; ns, non-significant). Horizontal lines indicate mean values. (J, K) Stage 16 SGs stained for GFP (for ManII-GFP) and WGA. Magnified images are shown for the boxed regions. Yellow arrows in J, ManII-GFP, and WGA puncta colocalize in WT SG. Asterisks in K indicate dispersed ManII-GFP signals in Papss mutants. Cyan arrows in K, little colocalization of ManII-GFP and WGA in the Papss SG. (L, L’) Line intensity profile showing the signal intensity of ManII-GFP (green) and WGA (magenta) along the yellow and cyan dotted lines in J and K, respectively. (M, N) Transmission electron microscopy (TEM) images of the Golgi. Yellow arrows in M, Golgi structures in wild-type (WT) SG cells. Magenta arrows in N, Golgi structures in the Papss SG. Cyan arrows in N, electron-dense structures in the Golgi of Papss mutant SG cells. (O, P) Stage 16 SGs immunostained for Rab11 and E-cadherin (Ecad). Magnified images are shown for boxed regions. Rab11 signals at the apical membrane are increased in Papss mutants (cyan arrowheads in P) compared to WT (yellow arrowheads in O), with occasional large punctate signals mislocalized to the basolateral domain (cyan arrow in P). (Q, R) Stage 16 SGs immunostained for Sec15 and Ecad. Compared to strong apical Sec15 signals in the WT SG (yellow arrowheads in Q), Sec15 signals are reduced in Papss mutants (cyan arrowheads in R). (S, T) Quantification of the intensity of Rab11 (S) and Sec15 (T) at the apical membrane regions in WT and Papss mutant SGs. Welch’s t-test (**p<0.05). Horizontal lines indicate mean values. (U, V) TEM images for the apical region of SG cells. Yellow arrowheads, electron-dense vesicles in both genotypes. Red arrowheads, empty, larger vesicle-like structures in Papss mutants. (W) Quantification of the electron-dense vesicle numbers per cell slice. Welch’s t-test (**p<0.05). Horizontal lines indicate mean values.
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Figure 3—source data 1
Raw data used to quantify the number and intensity of wheat germ agglutini (WGA).
See Figure 3I and I’ for the graphs.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig3-data1-v1.xlsx
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Figure 3—source data 2
Raw data used to test the colocalization of wheat germ agglutini (WGA) and ManII-GFP signals.
See Figure 3L and L’ for the graphs.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig3-data2-v1.xlsx
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Figure 3—source data 3
Raw data for the quantification of Rab11, Sec15, and Rab7 intensities, as well as the number of Rab7-positive puncta.
See Figure 3S and T, as well as Figure 3—figure supplement 3C and D, for graphs.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig3-data3-v1.xlsx
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Figure 3—source data 4
Raw data for the number of electron-dense secretory vesicles per transmission electron microscopy (TEM) slice are shown in Figure 3W.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig3-data4-v1.xlsx
Figure 3—figure supplement 1
The endoplasmic reticulum (ER) is intact in PAPS synthetase (Papss) mutants.
(A, B) Confocal images of stage 16 salivary glands (SGs) immunostained for GFP (for GFP-KDEL) and E-cadherin (Ecad). Magnified images are shown for the boxed regions. Yellow arrowheads, KDEL signals in the control (A) and Papss mutant (B) SGs. (C, D) Confocal images of stage 16 SGs immunostained for PH4ɑSG1 and ɑ-Spectrin. Magnified images are shown for boxed regions. Yellow arrows indicate PH4ɑSG1 signals in the ER in wild-type (WT) (C) and Papss mutant (D) SGs. (E, F) Transmission electron microscopy (TEM) images of SG epithelial cells. N indicates the nucleus. G indicates the Golgi. Yellow arrows indicate the ER.
Figure 3—figure supplement 2
Mutants of the Golgi component Grasp65 show an irregular salivary gland (SG) lumen phenotype.
(A–C) Confocal images of stage 16 SGs immunostained for E-cadherin (Ecad) and wheat germ agglutini (WGA). Yellow arrowheads in C, constrictions in the SG lumen of the Grasp65 mutant embryo.
Figure 3—figure supplement 3
Rab7 punctate number is decreased in PAPS synthetase (Papss) mutants.
(A) Confocal images of stage 16 salivary glands (SGs) immunostained for Rab7 and E-cadherin (Ecad). Magnified images are shown for boxed regions. Yellow arrowheads in A, Rab7 signals in wild-type (WT). Cyan arrowheads in B, Rab7 puncta are reduced in Papss mutants. (B) Quantification for the average mean gray value intensity of Rab7 puncta per sample. (C) Quantification of the average number of Rab7 puncta found in each cell for one sample. (B–C) Welch’s t-test (** p<0.01). Horizontal lines indicate mean values.
Figure 4
PAPS synthetase (Pappss) mutants show premature salivary gland (SG) cell death.
(A, B) Stage 16 SGs immunostained for CrebA, DCP-1, and E-cadherin (Ecad). Cross-sectional images are shown for the yellow dashed line in B. Yellow arrowheads indicate that DCP-1 signal is detected in the SG in Papss mutants. White dashed lines, the basal boundary of the SG. (C) Transmission electron microscopy (TEM) image of stage 15 SG cells, including a dying cell (yellow arrow). Cyan arrows, nuclei of normal living cells. (D) Quantification of the number of acellular regions in stage 16 SGs. (n=8 SGs, WT; n=10, Papss2; n=15, sage >Papss PD; Papss2; n=8, sage >Papss-PDK193A,F593P; Papss2) (E) Quantification of the number of SG cells at stage 16 (n=10 SGs, WT; n=10, Papss2; n=8, Papss2/Df; n=8, sage >p35; Papss2; n=10, sage >Papss PE; Papss2; n=8, sage >Papss-PDK193A,F593P; Papss2). One-way ANOVA test (****p<0.0001; ns, non-significant). (F, G) Stage 16 SGs immunostained for CrebA, Ecad, and mCherry (for Hemo3x-mCh). Cross-section images are shown for the yellow dashed lines. White dashed lines, the basal boundary of the SG. Yellow arrowhead in F, hemocyte signals outside of the control SG. White arrowhead in G, hemocyte signals are present inside the SG in Papss mutants. (H, I) Stage 16 SGs immunostained for DAPI, mCh (for Hemo3x-mCh), and Ndg. Magnified views are shown for boxed regions. Yellow dashed lines, the basal boundary of the SG. (H) In WT, hemocytes are found outside of the SG (yellow arrowheads), with Ndg forming a barrier at the basal boundary of the SG (yellow arrows). (I) In Papss mutants, hemocytes invade the SG epithelia (white arrowheads), where Nidogen (Ndg) signals are irregular or absent (white arrows).
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Figure 4—source data 1
Raw data for the number of acellular regions and of salivary gland (SG) cells shown in Figure 4D and E.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig4-data1-v1.xlsx
Figure 5 with 3 supplements
Loss of PAPS synthetase (Papss) disrupts the localization of Dumpy (Dpy) and Piopio (Pio) in the apical extracellular matrix (aECM).
(A) Stage 16 salivary glands (SGs) stained for a chitin-binding protein (CBP) and E-cadherin (Ecad). (B) Cartoon of the mCh-Pio fusion protein showing the region for the antibody and protease cleavage sites. (C, D) Stage 14 and stage 16 SGs immunostained for mCh (for mCh-Pio) and GFP (for Dpy-YFP). The mCh-Pio (white arrowheads) and Dpy-YFP (cyan arrowheads) signals are largely absent from the expanded region of the lumen. Along one side of the lumen, weak mCh-Pio signals colocalize with condensed Dpy-YFP signals (yellow arrows). (E-H’’’) Stage 14 and stage 16 SGs immunostained for Pio, GFP, and Ecad. Magnified images of boxed regions in E-H are shown in E’-H’’. F’’ and H’’’ are cross-sectional images of the SG lumen for regions indicated by white lines F’ and H’’. Yellow arrowheads, Pio and Dpy localized to the apical membrane. Cyan arrowheads, Dpy and Pio localized to the SG lumen. Note a higher proportion of Dpy and Pio localized to the lumen in the Papss mutant SG at stage 14 (cyan arrowheads in G’). White arrowheads in H’-H’’’, Dpy accumulates towards one side of the lumen in the stage 16 Papss SG. Yellow asterisks, Pio is absent from the lumen at stage 16 in the Papss SG.
Figure 5—figure supplement 1
Localization of Piopio (Pio) throughout salivary gland (SG) development.
(A) Confocal images of stage 11–16 SGs immunostained for Pio and E-cadherin (Ecad). Yellow arrowheads, Pio localizes to the SG lumen. Cyan arrowheads, Pio signals as cytoplasmic puncta. (B, C) Confocal images of stage 14 and 16 SGs immunostained for Pio and Ecad. Whereas filamentous luminal Pio signals are shown in control (B), no Pio signals are detected in pio null mutants (C).
Figure 5—figure supplement 2
Dumpy (Dpy) localization through salivary gland (SG) morphogenesis.
(A–B) Confocal images of stage 11–16 SGs immunostained using GFP and E-cadherin (Ecad). (A) Dpy-YFP heterozygous samples. Yellow arrowhead, Dpy is localized in the SG lumen in an organized, filamentous meshwork in Dpy-YFP/+. (B) Dpy-YFP homozygous samples. Cyan arrowheads, the SG lumen shows sites of constriction in Dpy-YFP/Dpy-YFP. White arrowheads, Dpy organization is irregular in the SG lumen of Dpy-YFP homozygous embryos. (C) Stage 16 Dpy-YFP/+SG stained for WGA. Luminal filamentous Dpy-YFP signals colocalize with wheat germ agglutini (WG) signals.
Figure 5—figure supplement 3
Quasimod (Qsm) localization patterns are not significantly affected in PAPS synthetase (Papss) mutants.
(A) Confocal images of stage 11–16 salivary glands (SGs) immunostained for GFP (for Dpy-YFP) and mCh (for mCh-Qsm). At stages 11–14, mCh-Qsm localized throughout the cytoplasm of SG cells, with increased localization at the apical membrane. At a late stage 14/early stage 15, the uniform cytoplasmic signals of mCh-Qsm began to decrease, and mCh-Qsm-positive vesicles formed near the apical domain and coalesced around SJs and the apical membrane. A few mCh-Qsm-positive vesicles were also observed in the basal region of SG cells. At stage 16, mCh-Qsm signals decreased in vesicles and instead increased at the apical membrane and were clearly detected at SJs, colocalizing with Dlg. Yellow asterisks, mCh-Qsm signals in the cytoplasm of SG cells. Yellow arrowheads, mCh-Qsm, show increased localization at the apical membrane and septate junctions in the late-stage SG. Cyan arrowheads, mCh-Qsm-positive puncta in the cytoplasm increase at early stage 15 in the SG. (B, C) Confocal images of stage 16 SGs immunostained for mCh and Dlg. Enlarged images are shown for boxed regions. mCh-Qsm partially colocalizes with Dlg in control (yellow arrowheads) and Papss mutant (cyan arrowheads) SGs. Yellow asterisks indicate dispersed mCh-Qsm signals near the apical membrane.
Figure 6 with 1 supplement
Piopio (Pio) is essential for a uniform salivary gland (SG) lumen diameter by maintaining Dumpy (Dpy) in the lumen.
(A, B) SGs at stages 13–16 stained for wheat germ agglutini (WGA) and E-cadherin (Ecad). Yellow arrowheads in A indicate filamentous luminal WGA signals. Cyan arrowheads in B, WGA signals at the apical membrane. Yellow asterisks indicate loss of lumina WGA signals. White arrowheads, constricted lumen. (C) Stage 16 SGs immunostained for Ecad and CrebA. Cross-sectional images for the yellow and green dotted lines are shown on the right. Yellow arrowhead, a flat protrusion of the lumen. Green arrowhead, constricted lumen. (D) Stage 16 SGs stained for Ecad and GFP (for Dpy-YFP). Projection images contain merged z-sections, from the apical surface of the SG cells to midway through the SG. Single section, one z-section at the approximate midpoint of the lumen width. Yellow arrowhead, area of constricted lumen. Yellow asterisk, luminal area with no Dpy-YFP signals. (E) Single z-sections of confocal images of SGs at stages 15 and 16 stained for Ecad, Pio, and WGA. White arrowheads, area of constricted lumen. Yellow asterisks, no luminal signals of WGA and Pio. (F) Quantification of the number of constrictions in the SG lumen at stage 16 (n=7 for both genotypes). (G, G’) SGs at stages 14–16 stained for Ecad, WGA, and Pio. Higher magnification of the yellow boxed regions is shown in G’. (H, I) Stage 16 SGs overexpressing NpWT (H) or Npmut (I) that are immunostained for GFP (for Dpy-YFP and Np-GFP), Pio, and mCh (for mCh-Pio). NpWT-overexpressing SGs show strong, uniform mCh-Pio signals in the lumen (cyan asterisks in H), whereas Npmut-overexpressing SGs show no detectable signals in the lumen (cyan asterisk in I). In NpWT-overexpressing SGs, the luminal mCh-Pio signals are so bright that the cytoplasmic, punctate mCh-Pio signals are rarely detectable alongside the luminal signals. Increasing the brightness to visualize the cytoplasmic punctate signals shows very strong luminal mCh-Pio signals (inset in H). NpWT overexpression shows high levels of Pio (magenta arrowheads in H) and dpy-YFP (green arrowheads in H) signals in both the apical membrane and lumen. In contrast, Npmut overexpression shows strong Pio signals in the apical membrane and various levels of Pio in the lumen. (I’, I’’) Higher magnification of the yellow boxed regions. The lumen diameter of the region with undetectable luminal Pio levels is slightly larger (magenta asterisk) compared to the region with clear luminal Pio signals (magenta arrowheads).
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Figure 6—source data 1
Raw data for the number of constrictions in the salivary gland (SG) of Piopio (pio) and Notopleura (Np) mutants.
see Figure 6F for the graph.
- https://cdn.elifesciences.org/articles/108292/elife-108292-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Loss of Piopio (pio) affects Dumpy (Dpy) localization patterns, and vice versa, in the salivary gland (SG).
(A, B) Confocal images of SGs immunolabeled for E-cadherin (Ecad) and wheat germ agglutini (WGA) at stages 13 and 14 in wild-type (WT) (A) and pio mutant (B) embryos. (C) Confocal images of pio mutant SGs from stages 12–15 immunostained for Ecad and GFP (for Dpy-YFP). Projection images contain merged z-sections, from the apical surface of the SG cells to midway through the SG. A single section from stage 16 at the approximate midpoint of the lumen width. (D) Confocal images of dpy mutant SG at stage 14 immunostained for Ecad, Pio, and WGA. Yellow boxes, area of higher magnification.
Figure 7
A proposed model.
(A) Cartoon diagram showing normal lumen expansion in wild-type (WT) salivary gland (SG) and defective luminal morphology in PAPS synthetase (Papss), Piopio (pio), and Notopleura (Np) mutants. (B) Organization of the zona pellucida (ZP) domain protein-containing apical extracellular matrix (aECM) in the SG. In WT, the ZP-C fragment of Pio (which is cleaved by Np and furin proteases) forms a complex with Dumpy (Dpy) to build a filamentous scaffold within the lumen. In Papss mutants, this fragment is absent from the lumen at stage 16, and the Dpy-containing aECM structure becomes aggregated and highly condensed. The absence of luminal Pio in pio and Np mutants results in the loss of luminal Dpy and lumen constrictions. Created with BioRender.com.
Additional files
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Supplementary file 1
A targeted Df screen revealed Papss as a key enzyme for SG lumen expansion.
- https://cdn.elifesciences.org/articles/108292/elife-108292-supp1-v1.docx
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Supplementary file 2
Fly strains used.
- https://cdn.elifesciences.org/articles/108292/elife-108292-supp2-v1.docx
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Supplementary file 3
Antibodies used.
- https://cdn.elifesciences.org/articles/108292/elife-108292-supp3-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/108292/elife-108292-mdarchecklist1-v1.docx
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Sulfation affects apical extracellular matrix organization during development of the Drosophila embryonic salivary gland tube
eLife 14:RP108292.
https://doi.org/10.7554/eLife.108292.3
