Localisation of Prickle in different mutant backgrounds

Prickle in a flamingo E59 clone

Loss of flamingo leads to a reduction of Prickle from the cell junctions, and a loss of asymmetry.

Fmi is required for junctional localisation of Prickle.

Confocal image of a 30-hour APF pupal wing containing a fmi^E59 mutant clone. The mutant cells are indicated with green dots. (B) LacZ marker for fmiE59 clone, (C) membrane-associated Pk.

Reference

Prickle mediates feedback amplification to generate asymmetric planar cell polarity signaling. D Tree et al., 2002

• Flybase reference

• Pubmed reference

Prickle in a frizzled^R52 mutant background in a pupal wing at 30h APF

Loss of frizzled disrupts Prickle junctional localisation and asymmetry.

Pk reduced at junctions and not asymmetric in absence of Frizzled.

Prickle is not asymmetrically localised in frizzled^R52 mutant wing cells.

Reference

Prickle mediates feedback amplification to generate asymmetric planar cell polarity signaling. D Tree et al., 2002

• Flybase reference

• Pubmed reference

Prickle in a dishevelled¹ mutant background

Prickle is not asymmetrically localised in dishevelled¹.

Loss of dishevelled disrupts Prickle asymmetry.

Dishevelled is required for asymmetric localisation and accumulation of Prickle as well as the other core polarity proteins.

Confocal images of pupal wings at 30 hours APF.

Reference

Prickle mediates feedback amplification to generate asymmetric planar cell polarity signaling. D Tree et al., 2002

• Flybase reference

• Pubmed reference

Prickle in a Diego mutant background in the eye

Unpublished localisation in the wing.

In the dgo³⁸⁰ mutant in the eye: Prickle localisation and pattern is unchanged.

The eye and the wing results seems to be different. In the eye the localisation of Pk seems unchanged. However, unpublished data in the Drosophila pupal wing suggests Diego is required for the asymmetric localisation of Prickle and the formation of puncta (unpublished).

Mutant tissue is marked by absence of β-gal staining or GFP (blue). White/yellow arrows indicate examples of clusters with PCP protein localisation in R3/R4 cell pair (equivalent to row 3 or 4).

Reference

Diego interacts with Prickle and Strabismus/Van Gogh to localize planar cell polarity complexes. G Das, A Jenny, T J Klein, S Eaton, and M Mlodzik, 2004

Pubmed reference

Prickle in a Strabismus mutant background

Prickle levels are increased at the junctions, Prickle localisation and asymmetry is lost in the absence of Strabismus.

In the absence of Strabismus, Prickle localises apical cell surface and asymmetry is lost.

Confocal images of pupal wings; distal is right and anterior is upwards. Mutant tissue indicated by absence of lacZ staining (red, left panels) strabismus⁶ mutant clones.

Reference

Strabismus is asymmetrically localised and binds to Prickle and Dishevelled during Drosophila planar polarity patterning. R Bastock, 2003

Strabismus promotes recruitment and degradation of Farnesylated Prickle in Drosophila melanogaster planar polarity specification. T-M Strutt, and Strutt, 2013

• Flybase reference

• Pubmed reference

Prickle in a Strabismus over-expression background

Loss of asymmetry of Prickle at the junctions.

Over-expression of Strabismus leads to a loss of Prickle asymmetry.

Clones of over-expression indicated by lacZ staining (red).

Prickle in Act-GAL4/UAS-Strabismus overexpression clone.

Reference

Strabismus is asymmetrically localised and binds to Prickle and Dishevelled during Drosophila planar polarity patterning. R Bastock, 2003

• Flybase reference

• Pubmed reference

Prickle in a Dishevelled over-expression background

Loss of asymmetry and an increase in the amount of Prickle at the junctions.

Over-expression of >>Dishevelled leads to a loss of Prickle asymmetry.

Act-GAL4/UAS-Dishevelled over-expression clone.

Clones of overexpression indicated by lacZ staining (red).

Reference

Strabismus is asymmetrically localised and binds to Prickle and Dishevelled during Drosophila planar polarity patterning. R Bastock, 2003

• Flybase reference

• Pubmed reference

Prickle in a Flamingo over-expression background

Loss of asymmetry and a decrease in the amount of Prickle at the junctions.

Over-expression of Flamingo leads to a loss of Prickle at the junctions and a loss of asymmetry.

Anti-Prickle antibody in Act-GAL4/UAS-Flamingo over-expression clone. UAS-Flamingo expresses the flamingo isoform that lacks the PBM Usui 1999. UAS-Flamingo clones in a wild type background stained for Prickle. Clones of over-expression indicated by lacZ staining (green).

White arrowheads point to clone boundaries, pupal wings are shown with distal to the right.

Reference

Differential stability of Flamingo protein complexes underlies the establishment of planar polarity. Strutt and Strutt, Current Biology, 2008

Pubmed reference

Prickle in a Frizzled over-expression background

Loss of Prickle anymmetry but an increase in Pk at the membrane.

Over-expression of Frizzled leads to a loss of Pk asymmetry (top half of the image).

Pk and Dsh colocalize in ptc-GAL4/UAS-fz wings (Pk, red; Dsh:GFP, green).

Reference

Prickle mediates feedback amplification to generate asymmetric planar cell polarity signaling. D Tree et al., 2002

• Flybase reference

• Pubmed reference

Prickle in a Diego over-expression background

Loss of Prickle asymmetry but an increase in large puncta.

Over-expression of Diego leads to a loss of Prickle asymmetry.

Act-GAL4/UAS-Diego over-expression clone.

Clones of over-expression indicated by lacZ staining (red).

Reference

Strabismus is asymmetrically localised and binds to Prickle and Dishevelled during Drosophila planar polarity patterning. R Bastock, 2003

• Flybase reference

• Pubmed reference

Prickle in a Flamingo over-expression background, mutant for Frizzled

Prickle localisation is not asymmetric in the absence of Fz.

Flamingo leads to a loss of Pk asymmetry but not a reduction in levels of Prickle.

There is no loss of Prickle in this background, this suggests that Frizzled is required for the removal of Prickle from the junctions when Flamingo is over-expressed.

What is interesting is that over-expression of Flamingo in the Frizzled mutant background, does not lead to more Pk at the junctions, which might be the case if Stbm and Pk were recruited preferentially to the boundaries in the absence of Frizzled.

White arrowheads point to clone boundaries, distal to the right.

Reference

Differential stability of Flamingo protein complexes underlies the establishment of planar polarity. Strutt and Strutt, Current Biology, 2008