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Long-range Notch-mediated tissue patterning requires actomyosin contractility

Ginger Hunter, Li He, Norbert Perrimon, Guillaume Charras, Edward Giniger, Buzz Baum

Posted on: 25 March 2018

Preprint posted on 2 February 2018

A recent preprint identifies a basal pool of actomyosin that is necessary for long-range Notch/Delta signalling underlying bristle patterning in the fruit fly

Selected by Yara E. Sánchez Corrales

Background

The pattern of bristles on the dorsal thorax (or notum) of the fruit fly Drosophila melanogaster is established by the transmembrane receptor Notch and its ligand Delta. During development, cells with low levels of Notch acquire sensory organ precursor (SOP) fate and become bristles, while cells expressing high levels of Notch retain epithelial fate. Since the activation of Notch is induced by contact with another cell expressing Delta, the process requires inhibition of neighbours’ cell fate. This patterning process is mediated both by neighbour-neighbour interactions and by long-range interactions via basal protrusions that extend the influence of cells beyond their immediate neighbours. Since Notch receptor could be mechanically activated, it was unclear whether the cytoskeleton on basal protrusions could play a role on bristle patterning.

Using quantitative live cell imaging and genetic manipulations, Hunter et al. (2018) identify a pool of non-muscle myosin II in the basal protrusions than mediate long-range Notch signaling, and establish a new role for actomyosin contractility during bristle patterning.

Key findings
The authors perturbed myosin II contractility by expressing a dominant-negative version of the heavy myosin chain, or a phospho-insensitive version of the myosin light chain, in specific cell populations (i.e only in signal-sending cells or in both signal-sending and receiving cells). They found that both adjacent and distant neighbours decrease the rate of Notch response compared to the control situation. Moreover, decreased myosin II activity caused defects in patterning: decreased spacing between SOP cells and poor row alignment (Figure 1D).

Figure 1. Decreased myosin contractility leads to defects in bristle precursor patterning. A) Notch response can be measured by the Notch reporter NsfGFP. B) The patterning involves cell-to-neighbour communication as well as long range interactions mediated by basal protrusions. D) Reduction of myosin contractility (sqhAA) leads to defects in the spacing of bristle precursors. Panels A and B are from Figure 1 and panel D is from Figure 2 of the original preprint.

Live imaging and particle image velocimetry (PIV) showed that basal protrusions were very dynamic and in constant flux, and their activity decreased if myosin II was compromised. These results suggest that the Notch response and subsequent patterning of bristles requires myosin II contractility.

How does actomyosin contractility contribute to the activity of Notch signalling? The authors showed that basal contacts were under tension. Thus protrusions may exert forces on one another as they moved that could contribute to activation of the Notch receptor along the protrusion.

What I like about this preprint and open questions

I found this work very interesting because it is an example of how signalling and patterning can be influenced by the contractile cytoskeleton. Typically, patterning is thought to act upstream of a developmental process that is “read” and “executed” by the cytoskeleton. Here, the authors show an example where the cytoskeleton itself is influencing patterning.

Another interesting point of the paper is the study of the 3D context of the cells. Until now, the majority of studies have focused at the role of myosin II at the apical domain, leaving behind the rest of the cell. In my opinion, the finding of very dynamic basal protrusions, which depend on actomyosin contractility, suggest that there might be interesting behaviours in 3D that need to be characterised. It would be very exciting to explore the basal dynamics in other types of epithelia.

The effect of decreased contractility in a phospho-insensitive version of the myosin light chain seems to affect patterning in different rows to various degree (Figure 1D, compare r1 and r3, for instance). It could be exciting to find out whether the dynamics of basal protrusions is comparable between rows. I am also curious about what effect an enhancement of contractility would have on the final pattern: would it cause closer bristles?

 

Tags: cell biology, development, notch, patterning

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