Measurements of mechanical properties of the blastula wall reveal which hypothesized mechanisms of primary invagination are physically plausible in the sea urchin Strongylocentrotus purpuratus.
Computer simulations showed that the elastic modulus of the cell layer relative to the elastic modulus of the extracellular layers predicted the effectiveness of different force-generating mechanisms for sea urchin primary invagination [L. A. Davidson, M. A. R. Koehl, R. Keller, and G. F. Oster (1995) Development 121, 2005-2018]. Here, we measured the composite elastic modulus of the cellular and extracellular matrix layers in the blastula wall of Strongylocentrotus purpuratus embryos at the mesenchyme blastula stage. Combined, these two layers exhibit a viscoelastic response with an initial stiffness ranging from 600 to 2300 Pa. To identify the cellular structures responsible for this stiffness we disrupted these structures and correlated the resulting lesions to changes in the elastic modulus. We treated embryos with cytochalasin D to disrupt the actin-based cytoskeleton, nocodazole to disrupt the microtubule-based cytoskeleton, and a gentle glycine extraction to disrupt the apical extracellular matrix (ECM). Embryos treated less than 60 min in cytochalasin D showed no change in their time-dependent elastic modulus even though F-actin was severely disrupted. Similarly, nocodazole had no effect on the elastic modulus even as the microtubules were severely disrupted. However, glycine extraction resulted in a 40 to 50% decrease in the elastic modulus along with a dramatic reduction in the hyalin protein at the apical ECM, thus implicating the apical ECM as a major mechanical component of the blastula wall. This finding bears on the mechanical plausibility of several models for primary invagination.
D000199:Actins / Q000502:physiology; D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D003198:Computer Simulation; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects; D004548:Elasticity; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000187:drug effects / Q000648:ultrastructure; D005998:Glycine / Q000494:pharmacology; D008433:Mathematics; D008870:Microtubules / Q000187:drug effects; D008954:Models, Biological; D009024:Morphogenesis / Q000502:physiology; D015739:Nocodazole / Q000494:pharmacology; D012617:Sea Urchins / Q000196:embryology; D014783:Viscosity