RESUMO
The chick limb bud has plasticity to reconstruct a normal skeletal pattern after a part of mesenchymal mass is excised to make a hole in its early stage of development. To understand the details of hole closure and re-establishment of normal limb axes to reconstruct a normal limb skeleton, we focused on cellular and molecular changes during hole repair and limb restoration. We excised a cube-shaped mass of mesenchymal cells from the medial region of chick hindlimb bud (stage 23) and observed the following morphogenesis. The hole had closed by 15 âh after excision, followed by restoration of the limb bud morphology, and the cartilage pattern was largely restored by 48 âh. Lineage analysis of the mesenchymal cells showed that cells at the anterior and posterior margins of the hole were adjoined at the hole closure site, whereas cells at the proximal and distal margins were not. To investigate cell polarity during hole repair, we analyzed intracellular positioning of the Golgi apparatus relative to the nuclei. We found that the Golgi apparatus tended to be directed toward the hole among cells at the anterior and posterior margins but not among cells at identical positions in normal limb buds or cells at the proximal and distal hole margins. In the manipulated limb buds, the frequency of cell proliferation was maintained compared with the control side. Tbx3 expression, which was usually restricted to anterior and posterior margins of the limb bud, was temporarily expanded medially and then reverted to a normal pattern as limb reconstruction proceeded, with Tbx3 negative cells reappearing in the medial regions of the limb buds. Thus, mesenchymal hole closure and limb reconstruction are mainly mediated by cells at the anterior and posterior hole margins. These results suggest that adjustment of cellular properties along the anteroposterior axis is crucial to restore limb damage and reconstruct normal skeletal patterns.
Assuntos
Padronização Corporal/fisiologia , Botões de Extremidades/citologia , Botões de Extremidades/embriologia , Células-Tronco Mesenquimais/metabolismo , Mesoderma/citologia , Mesoderma/embriologia , Esqueleto/embriologia , Animais , Proteínas Aviárias/metabolismo , Núcleo Celular/metabolismo , Polaridade Celular/fisiologia , Proliferação de Células/fisiologia , Embrião de Galinha , Extremidades/embriologia , Complexo de Golgi/metabolismo , Membro Posterior/embriologia , Transdução de Sinais/fisiologia , Esqueleto/citologia , Esqueleto/metabolismo , Proteínas com Domínio T/metabolismoRESUMO
It is now 50 years since Lewis Wolpert published the paper in which he set out the concept of Positional Information to explain how spatial patterns of cellular differentiation are generated. This concept has provided a universal model for pattern formation in embryonic development and regeneration and become part of the fabric of the field of developmental biology. Here I outline how Wolpert devised the concept of Positional Information and describe landmark studies from his lab investigating how Positional Information is specified in the developing chick limb.
Assuntos
Biologia do Desenvolvimento/métodos , Animais , Padronização Corporal/fisiologia , Embrião de Galinha , Extremidades/embriologia , Humanos , Morfogênese/fisiologiaRESUMO
To understand the morphogenetic mechanisms of organ development and regeneration, it is essential to clarify the inter-hierarchical relationship between microscopic, molecular/cellular activities and organ-level tissue deformation dynamics. While the former have been studied for several decades, the latter - macroscopic geometrical information about physical tissue deformation - is often missing, especially for many vertebrates. This is mainly because live recording of detailed cell behaviors in whole tissues during vertebrate organogenesis is technically difficult. In this study, we have developed a novel method that combines snapshot lineage tracing with Bayesian statistical estimation to construct whole-organ deformation maps from landmark data on limited numbers of space-time points. Following the validation of the method using artificially generated data sets, we applied it to the analysis of tissue deformation dynamics in chick limb development. A quantitative tissue deformation map for St.23-St.24 has been constructed, and its precision has been proven by evaluating its predictive performance. Geometrical analyses of the map have revealed a spatially heterogeneous volume growth pattern that is consistent with the expression pattern of a major morphogen and anisotropic tissue deformation along an axis. Thus, our method enables deformation dynamics analysis in organogenesis using practical lineage marking techniques.
Assuntos
Desenvolvimento Embrionário/fisiologia , Modelos Biológicos , Organogênese/fisiologia , Animais , Conjuntos de Dados como Assunto , HumanosRESUMO
Lewis Wolpert was a brilliant and inspiring scientist who made hugely significant contributions which underpin and influence our understanding of developmental biology today. He spent his career interested in how the fertilised egg can give rise to the whole embryo (and ultimately the adult) with one head, two arms, two legs, all its organs and importantly how cells become different from each other and how they 'know' what to become. His ideas revolutionised the way developmental biology was perceived and also reinvigorated, in particular, the key question of how pattern formation in embryonic development is achieved. He published over 200 scientific articles and received many accolades over his career for his work and services to science in the UK. These included a CBE (Commander of the Order of the British Empire) from the Queen, being elected a Fellow of the Royal Society and a Fellow of the Royal Society of Literature. He was also a recipient of the Waddington Medal from the British Society for Developmental Biology and was awarded The Royal Society's top honour, the Royal Medal in 2018. Lewis was also a gifted teacher and communicator, including being the author of a textbook on developmental biology used around the world to train the next generation of developmental biologists. This contribution was recognised in 2003, by the award of the Viktor Hamburger Outstanding Educator Award from the Society of Developmental Biology in the USA. Lewis always enjoyed giving talks and lectures, having an infectious and persuasive enthusiasm coupled with a sharp sense of humour. He also published articles in popular science journals (aimed at the public) such as New Scientist, Scientific American and The Scientist. Lewis also wrote several popular science books. He was a passionate advocate for the public understanding of science and was the Chair of The Royal Society/Royal Institution/British Association for the Advancement of Science Committee for Public Understanding of Science (1994-1998). For this contribution he was awarded The Royal Society Michael Faraday Medal for "excellence in communicating science to UK audiences". He presented the prestigious Royal Institution Christmas Lectures in 1986 entitled 'Frankenstein's Quest: development of life'. These lectures, six in total, are presented by leading scientists and aimed at the general public and broadcast on national television. On a personal level, Lewis influenced all who came into contact with him, shaped his students and postdocs careers and instilled in them, and the community as whole, a life-long love of developmental biology.
Assuntos
Distinções e Prêmios , Pessoal de Educação , Humanos , Estudantes , Estados Unidos , RedaçãoRESUMO
Homeoproteins have been shown to be expressed in a position-specific manner along the anterior-posterior axis in the developing chick feather bud, as seen also in the developing limb bud. These facts raise the possibility that there may be common mechanistic features in the establishment of the anterior-posterior polarity between both organs. In order to investigate this possibility, feather bud tissues were transplanted into the anterior region of limb buds to determine whether feather bud tissues possess properties such as the zone of polarizing activity of the limb bud. The manipulated limb bud formed a mirror image duplication of the skeletal elements, mainly (2)2234 digit pattern or sometimes 3(2)234. Both the anterior and posterior halves of feather bud tissue exhibited almost equal activity in inducing ectopic skeletal elements. Hoxd-12 and Hoxa-13 were expressed coordinately around the transplanted site of the operated limb bud. This secondary axis-inducing activity of the feather bud was enhanced when grafts were pretreated with trypsin. In contrast, the presumptive feather bud tissue and inter-feather bud tissue did not induce a secondary axis of the limb bud. These results suggest that the feather bud contains a region that exerts polarizing activity and that this region may play key roles in the formation of the anterior-posterior and, if it exists, proximal-distal axis of the feather bud, possibly via the regulation of region specific expression of Hox genes.
RESUMO
The ectodermal-mesenchymal interspace of the chick leg bud was studied at stages leading to the formation of the apical ectodermal ridge (A.E.R.) (stages 14 to 19 HH), using scanning and transmission electron microscopy. The main findings were: 1. a continuous basal lamina under the ectoderm; 2. extracellular fibrils interconnecting the basal lamina and mesenchymal cell processes; 3. an increase in the number of the fibrils during these stages, with the highest number under the A.E.R.; 4. branching mesenchymal cell processes that spread over the basal lamina, making contact with it in all stages. The morphology of the interspace and the changes in it suggest that extracellular material may be significant in the ectodermal-mesenchymal interactions in the limb bud.