The occipital lateral plate mesoderm is a novel source for vertebrate neck musculature

In vertebrates, body musculature originates from somites, whereas head muscles originate from the cranial mesoderm. Neck muscles are located in the transition between these regions. We show that the chick occipital lateral plate mesoderm has myogenic capacity and gives rise to large muscles located in the neck and thorax. We present molecular and genetic evidence to show that these muscles not only have a unique origin, but additionally display a distinct temporal development, forming later than any other muscle group described to date. We further report that these muscles, found in the body of the animal, develop like head musculature rather than deploying the programme used by the trunk muscles. Using mouse genetics we reveal that these muscles are formed in trunk muscle mutants but are absent in head muscle mutants. In concordance with this conclusion, their connective tissue is neural crest in origin. Finally, we provide evidence that the mechanism by which these neck muscles develop is conserved in vertebrates

The hypaxial origin of the epaxially located rhomboid muscles

In vertebrates, skeletal muscles of the body are made up of epaxial and hypaxial muscles based on their innervation and relative position to the vertebral column. The epaxial muscles are innervated by the dorsal branches of the spinal nerves and comprise the intrinsic (deep) back muscles, while the hypaxial muscles are innervated by the ventral branches of the spinal nerves including the plexus and consist of a heterogeneous group of intercostal, abdominal, and limb as well as girdle muscles. The canonical view holds that the epaxial muscles are derived from the medial halves of the somites, whereas the hypaxial muscles are all derived from the lateral somitic halves. The rhomboid muscles are situated dorsal to the vertebral column and therefore in the domain typically occupied by epaxial muscles. However, they are innervated by a ventral branch of the brachial plexus called the N. dorsalis scapulae. Due to the apparent inappropriate position of the muscle in relation to its innervation we investigated its origin to help clarify this issue. To study the embryonic origin of the rhomboid muscles, we followed derivatives of the medial and lateral somite halves using quail-chick chimeras. Our results showed that the rhomboid muscles are made up of cells derived mainly from the lateral portion of the somite. Therefore the rhomboid muscles which lie within the epaxial domain of the body, originate from the hypaxial domain of the somites. However, their connective tissue is derived from both medial and lateral somite

Commitment of chondrogenic precursors of the avian scapula takes place after epithelial-mesenchymal transition of the dermomyotome

<p>Abstract</p> <p>Background</p> <p>Cells of the epithelially organised dermomyotome are traditionally believed to give rise to skeletal muscle and dermis. We have previously shown that the dermomyotome can undergo epithelial-mesenchymal transition (EMT) and give rise to chondrogenic cells, which go on to form the scapula blade in birds. At present we have little understanding regarding the issue of when the chondrogenic fate of dermomyotomal cells is determined. Using quail-chick grafting experiments, we investigated whether scapula precursor cells are committed to a chondrogenic fate while in an epithelial state or whether commitment is established after EMT.</p> <p>Results</p> <p>We show that the hypaxial dermomyotome, which normally forms the scapula, does not generate cartilaginous tissue after it is grafted to the epaxial domain. In contrast engraftment of the epaxial dermomyotome to the hypaxial domain gives rise to scapula-like cartilage. However, the hypaxial sub-ectodermal mesenchyme (SEM), which originates from the hypaxial dermomyotome after EMT, generates cartilaginous elements in the epaxial domain, whereas in reciprocal grafting experiments, the epaxial SEM cannot form cartilage in the hypaxial domain.</p> <p>Conclusions</p> <p>We suggest that the epithelial cells of the dermomyotome are not committed to the chondrogenic lineage. Commitment to this lineage occurs after it has undergone EMT to form the sub-ectodermal mesenchyme.</p

The dermomyotome ventrolateral lip is essential for the hypaxial myotome formation

Background The myotome is the primitive skeletal muscle that forms within the embryonic metameric body wall. It can be subdivided into an epaxial and hypaxial domain. It has been shown that the formation of the epaxial myotome requires the dorsomedial lip of the dermomyotome (DML). Although the ventrolateral lip (VLL) of the dermomyotome is believed to be required for the formation of the hypaxial myotome, experimentally evidence for this statement still needs to be provided. Provision of such data would enable the resolution of a debate regarding the formation of the hypaxial dermomyotome. Two mechanisms have been proposed for this tissue. The first proposes that the intermediate dermomyotome undergoes cellular expansion thereby pushing the ventral lateral lip in a lateral direction (translocation). In contrast, the alternative view holds that the ventral lateral lip grows laterally. Results Using time lapse confocal microscopy, we observed that the GFP-labelled ventrolateral lip (VLL) of the dermomyotome grows rather than translocates in a lateral direction. The necessity of the VLL for lateral extension of the myotome was addressed by ablation studies. We found that the hypaxial myotome did not form after VLL ablation. In contrast, the removal of an intermediate portion of the dermomyotome had very little effect of the hypaxial myotome. These results demonstrate that the VLL is required for the formation of the hypaxial myotome. Conclusion Our study demonstrates that the dermomyotome ventrolateral lip is essential for the hypaxial myotome formation and supports the lip extension model. Therefore, despite being under independent signalling controls, both the dorsomedial and ventrolateral lip fulfil the same function, i.e. they extend into adjacent regions permitting the growth of the myotome

Use of AMBR250 as a small scale model for manufacturing-scale single-use bioreactors

Quality by Design (QbD) has become an integral part of biopharmaceutical process development and manufacturing. To gain the enhanced process understanding required by QbD, a well-designed small scale model that accurately predicts behavior at manufacturing scale is essential. This process understanding should ideally be achieved with rapid, efficient experimentation to decrease both the time and cost required for development. The ambr250 automated microscale bioreactor system has the potential to address all of these challenges. By embedding the ambr250 into the upstream process development workflow, throughput can be dramatically increased allowing for greater exploration of parameter operating ranges and more complete process understanding. However, the value of such microscale technologies hinges on their ability to accurately mimic manufacturing scale. We embarked on a study to demonstrate the applicability of the ambr250 (250 mL) as a small scale model for a 2000-L single-use bioreactor (SUB). We evaluated consistency of cell culture process performance from the ambr250 to 2000-L SUB scale along with intermediate scales such as our legacy small scale model (3-L glass stirred-tank reactors) and 50-L to 1000-L SUBs. Scalability was assessed using two monoclonal antibody molecules expressed from different CHO hosts (CHO K1 and DG44) and cultivated in different media platforms (chemically-defined and yeastolate-containing) to ensure broad applicability of the small scale model. Engineering principles were applied to develop appropriate agitation and gassing strategies at each scale to ensure comparability, with a power input based scaling strategy performing the best. Based on both univariate and multivariate data analysis methods the ambr250 behaved comparably to both our legacy small scale model and the SUBs for the assets evaluated. Areas of focus to further refine the ambr250 as a small scale model have also been identified

True 3D imaging with monocular cues using holographic stereography

A quantitative condition is derived to evaluate the monocular accommodation in holographic stereograms. We find that the reconstruction can be viewed as true-3D image when the whole scene is located in the monocular cues area, with compatible monocular cues and binocular cues. In contrast, it reveals incorrect monocular cues in the visible multi-imaging area and the lacking information area. To demonstrate our theoretical predictions, a pupil-function integral imaging algorithm is developed to simulate the mono-eye observation, and a holographic printing system is set up to fabricate the full-parallax holographic stereogram. Both simulation and experimental results match our theoretical predictions.Comment: 16 pages, 4 figure