XB-ART-48892PLoS One January 1, 2014; 9 (4): e95670.
Biomechanics and the thermotolerance of development.
Successful completion of development requires coordination of patterning events with morphogenetic movements. Environmental variability challenges this coordination. For example, developing organisms encounter varying environmental temperatures that can strongly influence developmental rates. We hypothesized that the mechanics of morphogenesis would have to be finely adjusted to allow for normal morphogenesis across a wide range of developmental rates. We formulated our hypothesis as a simple model incorporating time-dependent application of force to a viscoelastic tissue. This model suggested that the capacity to maintain normal morphogenesis across a range of temperatures would depend on how both tissue viscoelasticity and the forces that drive deformation vary with temperature. To test this model we investigated how the mechanical behavior of embryonic tissue (Xenopus laevis) changed with temperature; we used a combination of micropipette aspiration to measure viscoelasticity, electrically induced contractions to measure cellular force generation, and confocal microscopy to measure endogenous contractility. Contrary to expectations, the viscoelasticity of the tissues and peak contractile tension proved invariant with temperature even as rates of force generation and gastrulation movements varied three-fold. Furthermore, the relative rates of different gastrulation movements varied with temperature: the speed of blastopore closure increased more slowly with temperature than the speed of the dorsal-to-ventral progression of involution. The changes in the relative rates of different tissue movements can be explained by the viscoelastic deformation model given observed viscoelastic properties, but only if morphogenetic forces increase slowly rather than all at once.
PubMed ID: 24776615
PMC ID: PMC4002435
Article link: PLoS One
Genes referenced: actl6a dtl fn1 tbxt.2
Article Images: [+] show captions
|Figure 2. Blastopore closure at high and low temperatures.(A) Upper: vegetal view of an embryo showing the blastopore soon after the start of dorsal superficial involution. Lower left: kymograph of blastopore closure at 26°C, taken along the yellow line from the dorsal side to the ventral side, showing the points when dorsal (DI) and ventral (VI) superficial involution begin, and when the blastopore closes (BC). Right: kymograph taken along a line from the dorsal to the ventral side at 16°C. (B) The ratio (RCP) of the time for blastopore closure to the time for dorsal-to-ventral progression of involution versus the time (tP) for dorsal-to-ventral progression of involution. Dots indicate individual embryos. X's indicate medians for clutches (4 to 8 embryos each).|
|Figure 3. Comparison of viscoelastic models of morphogenesis for ramped versus stepped forces.(A) Diagram of model. Summed contractions (wavy lines) average out to stepped or ramped stresses (σ) depending on when cells begin contracting. When applied to the viscoelastic material with compliance J[t], the deformations (strains, ε) follow the time course of ramped forces more closely than stepped force. This can be visualized as adding up strains due to a series of stepped forces applied over time (dotted lines on right). (B) Predictions for RCP, the ratio of the time for morphogenesis (blastopore closure) to the time for patterning (D-V progression of involution), as a function of the time for patterning at temperature T, normalized to the time for patterning at 16°C, for ramped vs. stepped models for different values of β. Yellow dots: grand mean of experimentally observed values. The curves automatically converge to the right hand dot (at 16°C) where T2 = T1 since tC at T1 is used to calculate RCP at T2. (C) Comparison of the observed RCP at 26°C to the predictions for models with ramped or stepped forces, and with temperature invariant or varying mechanical properties (inset: prediction for stepped force model with temperature dependent mechanical properties on a log scale.) Error bars indicate confidence intervals. (D) Histogram of bootstrap resampling estimates of RCP at 26°C for each model (10,000 resamples total).|
|Figure 4. Duration of actomyosin contractions depends on temperature.(A) Sequential frames from a representative time-lapse sequence collected from the basal cortex of an animal cap ectoderm explant cultured on fibronectin-coated glass substrate. F-actin dynamics are revealed in cells expressing the actin-binding domain from moesin coupled to EGFP (moe-GFP) (left column). This sample collected at 16°C. (A′) Schematic of frames matching those in (A) highlighting the cell outline (dotted line) and hexagonal regions of the cell cortex identified as “F-actin contractions.” Regions are categorized as contractions when their integrated intensities are 50% greater than the mean intensity of the basal cell cortex. (B) Duration of individual F-actin contractions across the three temperature regimes. (C) Frequency distribution of the duration F-actin contractions at three temperatures. Note abundant short duration contractions at the low temperature regime.|