Odd-paired controls frequency doubling in Drosophila segmentation by altering the pair-rule gene regulatory network
- University of Cambridge, United Kingdom
Figures
Figure 1
The evolution of pair-rule patterning predates the evolution of long germ segmentation.
(A) Single segment periodicity is ancestral in arthropod segmentation, being found in spiders, millipedes, crustaceans and some insects (Davis et al., 2005; Pueyo et al., 2008). 'Pair-rule' …
Figure 2
Representative double fluorescent in situ hybridisation data for three combinations of pair-rule genes.
This figure shows a small subset of our wild-type dataset. Each column represents a different pairwise combination of in situ probes, while each row shows similarly-staged embryos of increasing …
Figure 3
Three phases of pair-rule gene expression, usually mediated by different classes of regulatory element.
(A) Representative expression patterns of each of the seven pair-rule genes at phase 1 (early cellularisation), phase 2 (mid cellularisation) and phase 3 (gastrulation). Pair-rule genes are …
Figure 4 with 2 supplements
Frequency-doubling of pair-rule gene expression patterns is almost simultaneous and coincides with the first expression of the segment-polarity genes.
Each row shows the expression of a particular pair-rule gene or segment-polarity gene, while each column represents a particular developmental timepoint. Late phase 2 and early phase 3 both …
Figure 4—figure supplement 1
Relative expression of pair-rule genes during frequency-doubling.
Each row shows the relative expression of two pair-rule genes, while each column represents a particular developmental timepoint. Late phase 2 and early phase 3 both correspond to late Bownes stage …
Figure 4—figure supplement 2
Relative expression of segment-polarity genes and pair-rule genes during frequency-doubling.
Each row shows the relative expression of a particular pair-rule gene and segment-polarity gene combination, while each column represents a particular developmental timepoint. Late phase 2 and early …
Figure 5
Odd does not repress prd transcription until phase 3.
Relative expression of prd and odd is shown in a late phase 2 embryo (just prior to frequency doubling) and an early phase 3 embryo (showing the first signs of frequency doubling). (A, B) Whole …
Figure 6
Schematic diagram of the transition to single-segment periodicity.
Schematic diagram showing segmentation gene expression at late phase 2 (A), early phase 3 (B) and late phase 3 (C). The horizontal axis represents an idealised portion of the AP axis (~12 nuclei …
Figure 7 with 1 supplement
Spatiotemporal expression of opa relative to odd.
Expression of opa relative to odd from early cellularisation until mid germband extension. (A) phase 1, lateral view; (B) early phase 2; (C–E) late phase 2; (F) early phase 3; (G, H) gastrulation; (I…
Figure 7—figure supplement 1
The cellular localisation of opa transcripts changes over the course of segmentation.
Relative expression of opa and ftz is shown in embryos at phase 1, phase 2 and phase 3. (A–C) Whole embryos, lateral view, anterior left, dorsal top. Single-channel images are shown in greyscale …
Figure 8 with 6 supplements
Pair-rule gene expression is perturbed from gastrulation onwards in opa mutant embryos.
Pair-rule gene expression in wild-type and opa mutant embryos at late cellularisation, late gastrulation, and early germband extension. During cellularisation, pair-rule gene expression in opa …
Figure 8—figure supplement 1
Pair-rule gene expression in opa mutant embryos at cellularisation.
Relative expression patterns of pair-rule genes in wild-type and opa mutant embryos at late cellularisation. All images are double in situs for odd and one other pair-rule gene. Individual channels …
Figure 8—figure supplement 2
Pair-rule gene expression in opa mutant embryos at gastrulation.
Relative expression patterns of pair-rule genes in wild-type and opa mutant embryos at gastrulation. All images are double in situs for odd and one other pair-rule gene. Individual channels are …
Figure 8—figure supplement 3
Pair-rule gene expression in opa mutant embryos at early germband extension.
Relative expression patterns of pair-rule genes in wild-type and opa mutant embryos at early germband extension. All images are double in situs for odd and one other pair-rule gene. Individual …
Figure 8—figure supplement 4
The transition to single-segment periodicity does not occur in opa mutant embryos.
Comparison of early phase 3 segmentation gene expression in wild-type and opa mutant embryos. The horizontal axis represents an idealised portion of the AP axis (~12 nuclei across). The grey …
Figure 8—figure supplement 5
Opa activates the eve “'late' element.
eve and odd expression in wild-type and opa mutant embryos at various timepoints spanning mid-phase 2 (mid-cellularisation, top row) to late phase 3 (onset of germband extension, bottom row). In opa …
Figure 8—figure supplement 6
'Late' eve expression is observed in cells that do not express prd.
eve and prd expression in wild-type embryos during phase 3. During early phase 3 (left), eve is strongly expressed in stripes ~2 cells wide. These stripes only partially overlap with the 'P' stripes …
Figure 9
Opa-dependent expression pattern changes are delayed in opa hypomorphic embryos.
Expression of selected pair-rule genes compared between embryos wild-type, hypomorphic (opa5), or null mutant (opa8) for opa. Arrowheads mark evidence of Opa-dependent regulatory interactions in opa5…
Figure 10 with 1 supplement
Model for the Opa-dependent patterning of the even-numbered parasegment boundaries.
(A) Schematic showing the phasing of odd, slp and en relative to Runt and Ftz protein at phase 3. The horizontal axis represents part of a typical double-segment pattern repeat along the AP axis of …
Figure 10—figure supplement 1
Segment-polarity gene expression in opa mutant embryos.
Development of segment-polarity gene expression in wild-type and opa mutant embryos. Arrowheads marks segment-polarity stripes that normally contribute to odd-numbered parasegment boundaries …
Appendix 1—figure 1 with 1 supplement
Expression of odd at phase 2 versus phase 3.
Relative expression of odd and other pair-rule genes (A, B – eve; C, D – hairy; E, F – runt; G, H - slp) is shown in late phase 2 embryos (A, C, E, G) and in early phase 3 embryos (B, D, F, H). …
Appendix 1—figure 1—figure supplement 1
Model for the regulation of odd transcription at phase 2 versus phase 3.
Model for the differential regulation of odd expression by pair-rule proteins at late phase 2 (A, B) versus mid phase 3 (C, D). (A, C) Schematic diagrams showing the expression of odd relative to …
Appendix 1—figure 2 with 2 supplements
Expression of slp at phase 2 versus phase 3.
Relative expression of slp and other pair-rule genes (A, B – eve; C, D – runt; E, F – ftz; G, H - odd) is shown in late phase 2 embryos (A, C, E, G) and in early phase 3 embryos (B, D, F, H). …
Appendix 1—figure 2—figure supplement 1
Model for the regulation of slp transcription at phase 2 versus phase 3.
Model for the differential regulation of slp expression by pair-rule proteins at late phase 2 (A, B) versus mid phase 3 (C, D). (A, C) Schematic diagrams showing the expression of slp relative to …
Appendix 1—figure 2—figure supplement 2
Runt represses slp during phase 2 in both wild-type and ftz mutant embryos.
Relative expression of runt and slp in wild-type and ftz mutant embryos. In both cases, co-expression of runt and slp is not seen until phase 3. Individual channels are shown to the right of each …
Appendix 1—figure 3 with 1 supplement
Expression of runt at phase 2 versus phase 3.
Relative expression of runt and other pair-rule genes (A, B – hairy; C, D – odd; E, F – eve; G, H - ftz) is shown in late phase 2 embryos (A, C, E, G) and in early phase 3 embryos (B, D, F, H). …
Appendix 1—figure 3—figure supplement 1
Model for the regulation of runt transcription at phase 2 versus phase 3.
Model for the differential regulation of runt expression by pair-rule proteins at late phase 2 (A, B) versus early phase 3 (C, D). (A, C) Schematic diagrams showing the expression of runt relative …
Appendix 2—figure 1 with 2 supplements
The ftz/odd anterior boundary offsets are lost in opa mutant embryos at gastrulation.
Relative expression of ftz and odd in wild-type and opa mutant embryos. (A–D) Whole embryos, anterior left; (A–C) show lateral views, (D) shows a ventral view. Single channels are shown to the right …
Appendix 2—figure 1—figure supplement 1
The odd primary stripes expand anteriorly in opa mutant embryos.
Relative expression of eve and odd at early phase 3 in wild-type and opa mutant embryos. (A, B) Whole embryos, lateral view, anterior left, dorsal top. Individual channels are shown to the right of …
Appendix 2—figure 1—figure supplement 2
Model for the patterning of the anterior boundaries of ftz and odd.
Model for the regulation of ftz and odd expression by Eve and Runt, in both the absence (A, B) and the presence (C, D) of Opa protein. (A, C) Schematic diagrams showing the expression of ftz and odd …
Appendix 2—figure 2 with 3 supplements
odd stripe 7 expands anteriorly and ventrally in opa mutants.
Expression of odd relative to that of eve, runt and prd, in wild-type and opa mutant embryos. (A–J) Whole embryos, individual channels shown below each double channel image (odd right). (A, F) Late …
Appendix 2—figure 2—figure supplement 1
odd stripe 7 appears after the primary stripes of prd, but before the primary stripes of slp.
Expression of odd relative to that of prd and slp over the course of cellularisation. At early phase 2, prd expression in the trunk has appeared, and there are only 6 odd stripes. At mid phase 2, odd…
Appendix 2—figure 2—figure supplement 2
The posterior border of eve stripe 7 shifts anteriorly relative to the anterior border of odd stripe 7.
Expression of odd relative to that of runt and eve, in wild-type embryos at cellularisation (mid phase 2) and gastrulation (phase 3). (A–D) Whole embryos, lateral view, anterior left, dorsal top. …
Appendix 2—figure 2—figure supplement 3
Model for the patterning of the anterior boundaries of the odd primary stripes.
Schematic showing the phasing of odd expression relative to Eve, Runt and Opa protein. The horizontal axis represents part of a double-segment pattern repeat along the AP axis of the embryo. Black …
