Abstract

Connection of tubules into larger networks is the key process for the development of circulatory systems. In Drosophila development, tip cells of the tracheal system lead the migration of each branch and connect tubules by adhering to each other and simultaneously changing into a torus-shape. We show that as adhesion sites form between fusion cells, myosin and microtubules form polarized bundles that connect the new adhesion site to the cells' microtubule-organizing centres, and that E-cadherin and retrograde recycling endosomes are preferentially deposited at the new adhesion site. We demonstrate that microtubules help balancing tip cell contraction, which is driven by myosin, and is required for adhesion and tube fusion. We also show that retrograde recycling and directed secretion of a specific matrix protein into the fusion-cell interface promote fusion. We propose that microtubule bundles connecting these cell–cell interfaces coordinate cell contractility and apical secretion to facilitate tube fusion.

Highlights

  • Connection of tubules into larger networks is the key process for the development of circulatory systems

  • We found that the E-cadherin that accumulated at the new cell interface is not recycled from the cell surface, but is instead drawn from a newly synthesized pool and recruited preferentially to the FC contact site, and not to existing adherens junctions between FCs and stalk cells

  • We considered one possible mechanism of preferential deposition, in which microtubules transport endosomes containing E-cadherin towards the contact site and we attempted to test this model by imaging vesicular trafficking of the complex containing E-cadherin-green fluorescent protein (GFP) and other adherens junction components

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Summary

Introduction

Connection of tubules into larger networks is the key process for the development of circulatory systems. We propose that microtubule bundles connecting these cell–cell interfaces coordinate cell contractility and apical secretion to facilitate tube fusion. Some epithelial cells acquire the ability to migrate while retaining cell–cell adhesion, allowing these cells to migrate as a group[6,7,8,9] This is possible due to the polarized nature of the epithelia; the apical side retains the cell–cell interface, while migratory protrusions form on the basal cell surface. To examine how cell adhesion and cell contractility are coupled to establish a new cell interface, we examined the anastomosis formation in the Drosophila tracheal system In this process, a pair of tip cell (hereafter called fusion cell, FC) forms an adherens junction de novo (Fig. 1a,b) and deposit extracellular matrix materials into newly forming lumen. In addition to this cell-intrinsic contractile force, the preferential deposition of molecules via the apical secretion pathway promotes maturation of the matrix in the lumen that forms in the FC contact interface and helps to fuse the plasma membranes

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