SUMMARY Background Epithelial invagination is a fundamental morphogenetic behavior that transforms a flat cell sheet into a pit or groove. Previous studies of invagination have focused on the role of actomyosin-dependent apical contraction; other mechanisms remain largely unexplored. Results We combined experimental and computational approaches to identify a two-step mechanism for endoderm invagination during ascidian gastrulation. During Step 1, which immediately precedes invagination, endoderm cells constrict their apices due to Rho/Rhokinase-dependent apical enrichment of 1P–myosin. Our data suggest that endoderm invagination itself occurs during Step 2, without further apical shrinkage, via a novel mechanism we call collared rounding: Rho/Rho-kinase-independent lateral enrichment of 1P–myosin drives apico-basal shortening, while Rho/Rho-kinase-dependent enrichment of 1P and 2P myosin in circumapical collars is required to prevent apical expansion and for deep invagination. Simulations show that boundary-specific tension values consistent with these distributions of active myosin can explain the cell shape changes observed during invagination both in normal embryos and in embryos treated with pharmacological inhibitors of either Rho-kinase or Myosin II ATPase. Indeed, we find that the balance of strong circumapical and basolateral tension is the only mechanism based on differential cortical tension that can explain ascidian endoderm invagination. Finally, simulations suggest that mesectoderm cells resist endoderm shape changes during both steps and we confirm this prediction experimentally. Conclusions Our findings suggest that early ascidian gastrulation is driven by the coordinated apposition of circumapical and lateral endoderm contraction, working against a resisting mesectoderm. We propose that similar mechanisms may operate during other invaginations.

Unidirectional zippering is a key step in neural tube closure that remains poorly understood. Here, we combine experimental and computational approaches to identify the mechanism for zippering in a basal chordate, Ciona intestinalis. We show that myosin II is activated sequentially from posterior to anterior along the neural/epidermal (Ne/Epi) boundary just ahead of the advancing zipper. This promotes rapid shortening of Ne/Epi junctions, driving the zipper forward and drawing the neural folds together. Cell contact rearrangements (Ne/Epi + Ne/Epi → Ne/Ne + Epi/Epi) just behind the zipper lower tissue resistance to zipper progression by allowing transiently stretched cells to detach and relax toward isodiametric shapes. Computer simulations show that measured differences in junction tension, timing of primary contractions, and delay before cell detachment are sufficient to explain the speed and direction of zipper progression and highlight key advantages of a sequential contraction mechanism for robust efficient zippering.

The cuttlebone is a rigid buoyancy tank that imposes a depth limit on Sepia, the only living speciose cephalopod genus with a chambered shell. Sections of 59 cuttlebones from a geographically diverse sample of 11 species were examined using confocal microscopy. Sepia species that live at greater depths had thicker septa and less space between pillars than did shallow species. A plate theory analysis of cuttlebone strength based on these two measures predicted maximum capture depths accurately in most species. Thus cuttlebone morphology confers differing degrees of strength against implosion from hydrostatic pressure, which increases with increasing habitat depth. Greater strength may come at the cost of increased cuttlebone density, which impinges on the cuttlebone's buoyancy function.

There was an error published in Development 142, 1869-1878.On p. 1869, the sentence 'Recently, the JM domain was implicated in specifying the position of a supercellular actomyosin cable, by concentrating atypical protein kinase C (aPKC) away from the site of the cable, thereby allowing an increase in localized Rhomboid (Rho) activity (Roper, 2012).' should instead have read:'Recently, the JM domain was implicated in specifying the position of a supercellular actomyosin cable, by concentrating atypical protein kinase C (aPKC) away from the site of the cable, thereby allowing an increase in localized Rho-kinase (Rok) activity (Roper, 2012).'The publishers and authors apologise to readers for this mistake. 2226 ABSTRACTThe transmembrane protein Crumbs (Crb) functions in apical polarity and epithelial integrity. To better understand its role in epithelial morphogenesis, we examined Crb localization and dynamics in the late follicular epithelium of Drosophila. Crb was unexpectedly dynamic during middle-to-late stages of egg chamber development, being lost from the marginal zone (MZ) in stage 9 before abruptly returning at the end of stage 10b, then undergoing a pulse of endocytosis in stage 12. The reappearance of MZ Crb is necessary to maintain an intact adherens junction and MZ. Although Crb has been proposed to interact through its juxtamembrane domain with Moesin (Moe), a FERM domain protein that regulates the cortical actin cytoskeleton, the functional significance of this interaction is poorly understood. We found that whereas the Crb juxtamembrane domain was not required for adherens junction integrity, it was necessary for MZ localization of Moe, aPKC and F-actin. Furthermore, Moe and aPKC functioned antagonistically, suggesting that Moe limits Crb levels by reducing its interactions with the apical Par network. Additionally, Moe mutant cells lost Crb from the apical membrane and accumulated excess Crb at the MZ, suggesting that Moe regulates Crb distribution at the membrane. Together, these studies reveal reciprocal interactions between Crb, Moe and aPKC during cellular morphogenesis.

Newly settled marine invertebrates are typically less than 1 mm tall and face physical and biotic factors distinct from those facing adults, including a fluid environment dominated by high viscosity and deep boundary layers, and a size-specific suite of predators and competitors. If juveniles have distinct morphologies from adults, or morphologies that function differently at small scales, these differences could have important implications for juvenile performance. We compared the ontogenetic scaling of morphological parameters related to suspension feeding in 6 species of ascidians covering a wide taxonomic range. Many aspects of early juvenile morphology differed from those of conspecific adults, though not invariably in ways likely to improve feeding performance at small size. Some juvenile morphologies that are detrimental to efficient suspension feeding may serve other functions, whereas others are the result of commencing suspension feeding before the completion of metamorphosis. Thus, not all ascidian species appear specialized for higher rates of suspension feeding and early growth.