Isolation, culture, and differentiation of Blastema cells from the regenerating caudal fin of zebrafish

The caudal fin of teleost fish has become an excellent system for investigating the mechanisms of epimorphic regeneration. Upon amputation of the caudal fin, a mass of undi erentiated cells, called blastema, proliferate beneath the wound-epidermis and di erentiate into various cell types to faithfully restore the missing fin structures. Here we describe a protocol that can be used to isolate and culture blastema cells from zebrafish. Primary cultures were initiated from 36 h post-amputation (hpa) blastema and optimal cell growth was achieved using L-15 medium supplemented with 5% fetal bovine serum in plates either coated with fibronectin or uncoated. After seeding, zebrafish blastema cells formed a uniform culture and exhibited polygonal shapes with prominent nucleus, while various cell types were also observed after few days in culture indicating cell di erentiation. Upon treatment with all-trans retinoic acid, zebrafish blastema cells di erentiated into neuron-like and oligodendritic-like cells. Immunocytochemistry data also revealed the presence of mesenchymal and neuronal cells. The availability of blastema cell cultures could contribute to a better understanding of epimorphic regeneration by providing a mean to investigate the mechanisms underlying blastema cell di erentiation. Furthermore, this protocol is simple, rapid, and cost-e cient, and can be virtually applied to the development of any fish blastema cell culture.FP7/227799; UID/Multi/04326/2019info:eu-repo/semantics/publishedVersio

Les habitudes technologiques au cégep : résultats d'une enquête effectuée auprès de 30 724 étudiants

Texte en anglais.Bibliographie : pages 55-56

Dermoskeleton morphogenesis in zebrafish fins

Zebrafish fins have a proximal skeleton of endochondral bones and a distal skeleton of dermal bones. Recent experimental and genetic studies are discovering mechanisms to control fin skeleton morphogenesis. Whereas the endochondral skeleton has been extensively studied, the formation of the dermal skeleton requires further revision. The shape of the dermal skeleton of the fin is generated in its distal growing margin and along a proximal growing domain. In these positions, dermoskeletal fin morphogenesis can be explained by intertissue interactions and the function of several genetic pathways. These pathways regulate patterning, size, and cell differentiation along three axes. Finally, a common genetic control of late development, regeneration, and tissue homeostasis of the fin dermoskeleton is currently being analyzed. These pathways may be responsible for the similar shape obtained after each morphogenetic process. This provides an interesting conceptual framework for future studies on this topic. Developmental Dynamics 239:2779–2794, 2010. © 2010 Wiley-Liss, Inc

Banding pattern and fibrillogenesis of ceratotrichia in shark fins

Study by light microscopy, scanning electron microscopy and transmission electron microscopy of the distribution, structure and growth of ceratotrichia in the anterodorsal fin of a lemon shark, Negaprion brevirostris , and in the tailfin of a nurse shark, Ginglymostoma cirratum , shows that the ceratotrichia are large collagen fibers which develop in bilateral rows within the dermis. Surrounding each ceratotrichium is a layer of peritrichial fibroblasts containing secretory vesicles, which appear to be the source of matrix constituents. The peritrichial matrix contains bundles of fine, unbanded collagen fibrils as well as larger, banded fibrils like those in the matrix of ordinary connective tissue. The structure of the peritrichial fibroblasts and of the subjacent peritrichial matrix is the same as that of the fibroblasts and matrix of the conventional connective tissue throughout the fin dermis. Ceratotrichia grow by apposition of collagen fibrils from the peritrichial matrix. In cross section the ceratotrichia appear layered, evidently because of close packing of constituent fibrils in lamellae. In longitudinal section the ceratotrichia exhibit the conventional a, b, c, d and e bands of collagen. The e bands of show two distinct subbands, and the b bands three subbands. Periodicity of the banding pattern is approximately 640 Å like that of conventional collagen fibrils.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50267/1/1051540202_ftp.pd

The development and growth of tissues derived from cranial neural crest and primitive mesoderm is dependent on the ligation status of retinoic acid receptor γ:evidence that retinoic acid receptor γ functions to maintain stem/progenitor cells in the absence of retinoic acid

Retinoic acid (RA) signaling is important to normal development. However, the function of the different RA receptors (RARs)-RARα, RARβ, and RARγ-is as yet unclear. We have used wild-type and transgenic zebrafish to examine the role of RARγ. Treatment of zebrafish embryos with an RARγ-specific agonist reduced somite formation and axial length, which was associated with a loss of hoxb13a expression and less-clear alterations in hoxc11a or myoD expression. Treatment with the RARγ agonist also disrupted formation of tissues arising from cranial neural crest, including cranial bones and anterior neural ganglia. There was a loss of Sox 9-immunopositive neural crest stem/progenitor cells in the same anterior regions. Pectoral fin outgrowth was blocked by RARγ agonist treatment. However, there was no loss of Tbx-5-immunopositive lateral plate mesodermal stem/progenitor cells and the block was reversed by agonist washout or by cotreatment with an RARγ antagonist. Regeneration of the caudal fin was also blocked by RARγ agonist treatment, which was associated with a loss of canonical Wnt signaling. This regenerative response was restored by agonist washout or cotreatment with the RARγ antagonist. These findings suggest that RARγ plays an essential role in maintaining stem/progenitor cells during embryonic development and tissue regeneration when the receptor is in its nonligated state