scholarly journals Dissection of Xenopus laevis Neural Crest for in vitro Explant Culture or in vivo Transplantation

10.3791/51118 ◽  
2014 ◽  
Author(s):  
Cecile Milet ◽  
Anne Helene Monsoro-Burq
Keyword(s):  
Xenopus Laevis ◽  
Neural Crest ◽  
Explant Culture

In vitro studies on skeletogenic potential of membrane bone periosteal cells

Development ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 185-207
Author(s):  
Peter Thorogood
Keyword(s):  
Neural Crest ◽  
Explant Culture ◽  
General Context ◽  
Avian Embryo ◽  
Spatial And Temporal Patterns ◽  
Chondrogenic Potential ◽  
Membrane Bone ◽  
Periosteal Cells

In the avian embryo ectomesenchyme cells, derived from the mesencephalic level of the cranial neural crest, migrate into the presumptive maxillary region and subsequently differentiateinto the membrane bones and associated secondary cartilage of the upper jaw skeleton. The cartilage arises secondarily within the periosteum at points of articulation between membrane bones and provides an embryonic articulating surface. The stimulus for the differentiation of secondary cartilage is believed to be intermittent pressure and shear created at the developing embryonic movement. The development of one such system - the quadratojugal, has been analysed using organ and explant culture techniques and studied with particular reference to the differentiation of periosteal cells into secondary cartilage. A number of conclusions were reached. (1) Normally only cells at discrete loci express a chondrogenic potential ,in vivo: the periosteal cells at these sites of future articulation become committed to chondrogenesis during stage 35, more than 24 h before cartilage is identifiable ,in vivo. (2) However, cells with a ‘latent’ chondrogenic potential are widespread in membrane bone periosteum and occur over most, if not all, of the surface area of the bone. This potential is expressed in the ‘permissive’ environment created by submersion of the tissue in explant culture or in submerged organ culture. (3) This chondrogenic potential exists long before the time at which commitment of cartilage-forming cells occurs and even presumptive maxillary ectomesenchyme at stage 29 has a limited ability to form cartilage ,in vitro. It is suggested that spatial position is a principal factor controlling the differentiation of secondary cartilage. Ectomesenchyme cells with the potential to form secondary cartilage are widespread but it is only those cells whose migration from the neural crest positions them and their progeny at the site of a presumptive joint which subsequently express this potential. This epigenetic interpretation is discussed in the general context of development mechanisms underlying the spatial and temporal patterns in which neural crest-derived cells differentiate to produce bone and cartilage during the formation of the head skeleton.

scholarly journals Regulation of sarcomagenesis by the empty spiracles homeobox genes EMX1 and EMX2

2021 ◽  
Vol 12 (6) ◽  
Author(s):  
Manuel Pedro Jimenez-García ◽  
Antonio Lucena-Cacace ◽  
Daniel Otero-Albiol ◽  
Amancio Carnero
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Transcription Factors ◽  
Neural Crest ◽  
Tumor Suppressors ◽  
Homeobox Genes ◽  
Neuronal Cell ◽  
Cell Gene Expression

AbstractThe EMX (Empty Spiracles Homeobox) genes EMX1 and EMX2 are two homeodomain gene members of the EMX family of transcription factors involved in the regulation of various biological processes, such as cell proliferation, migration, and differentiation, during brain development and neural crest migration. They play a role in the specification of positional identity, the proliferation of neural stem cells, and the differentiation of certain neuronal cell phenotypes. In general, they act as transcription factors in early embryogenesis and neuroembryogenesis from metazoans to higher vertebrates. The EMX1 and EMX2’s potential as tumor suppressor genes has been suggested in some cancers. Our work showed that EMX1/EMX2 act as tumor suppressors in sarcomas by repressing the activity of stem cell regulatory genes (OCT4, SOX2, KLF4, MYC, NANOG, NES, and PROM1). EMX protein downregulation, therefore, induced the malignance and stemness of cells both in vitro and in vivo. In murine knockout (KO) models lacking Emx genes, 3MC-induced sarcomas were more aggressive and infiltrative, had a greater capacity for tumor self-renewal, and had higher stem cell gene expression and nestin expression than those in wild-type models. These results showing that EMX genes acted as stemness regulators were reproduced in different subtypes of sarcoma. Therefore, it is possible that the EMX genes could have a generalized behavior regulating proliferation of neural crest-derived progenitors. Together, these results indicate that the EMX1 and EMX2 genes negatively regulate these tumor-altering populations or cancer stem cells, acting as tumor suppressors in sarcoma.

scholarly journals Lamellipodin and the Scar/WAVE complex cooperate to promote cell migration in vivo

2013 ◽  
Vol 203 (4) ◽  
pp. 673-689 ◽  
Author(s):  
Ah-Lai Law ◽  
Anne Vehlow ◽  
Maria Kotini ◽  
Lauren Dodgson ◽  
Daniel Soong ◽  
...  
Keyword(s):  
Cell Migration ◽  
Neural Crest ◽  
Neural Crest Cell ◽  
Direct Interaction ◽  
Dependent Manner ◽  
Border Cell ◽  
Promote Cell Migration ◽  
Wave Complex

Cell migration is essential for development, but its deregulation causes metastasis. The Scar/WAVE complex is absolutely required for lamellipodia and is a key effector in cell migration, but its regulation in vivo is enigmatic. Lamellipodin (Lpd) controls lamellipodium formation through an unknown mechanism. Here, we report that Lpd directly binds active Rac, which regulates a direct interaction between Lpd and the Scar/WAVE complex via Abi. Consequently, Lpd controls lamellipodium size, cell migration speed, and persistence via Scar/WAVE in vitro. Moreover, Lpd knockout mice display defective pigmentation because fewer migrating neural crest-derived melanoblasts reach their target during development. Consistently, Lpd regulates mesenchymal neural crest cell migration cell autonomously in Xenopus laevis via the Scar/WAVE complex. Further, Lpd’s Drosophila melanogaster orthologue Pico binds Scar, and both regulate collective epithelial border cell migration. Pico also controls directed cell protrusions of border cell clusters in a Scar-dependent manner. Taken together, Lpd is an essential, evolutionary conserved regulator of the Scar/WAVE complex during cell migration in vivo.

The neural crest population responding to endothelin-3 in vitro includes multipotent cells

1997 ◽  
Vol 110 (14) ◽  
pp. 1673-1682 ◽  
Author(s):  
J.G. Stone ◽  
L.I. Spirling ◽  
M.K. Richardson
Keyword(s):  
Neural Crest ◽  
Schwann Cells ◽  
Cell Types ◽  
Developmental Potential ◽  
Colony Assay ◽  
Cellular Targets ◽  
Multipotent Cells ◽  
Endothelin 3

The peptide endothelin 3 (EDN3) is essential for normal neural crest development in vivo, and is a potent mitogen for quail truncal crest cells in vitro. It is not known which subpopulations of crest cells are targets for this response, although it has been suggested that EDN3 is selective for melanoblasts. In the absence of cell markers for different precursor types in the quail crest, we have characterised EDN3-responsive cell types using in vitro colony assay and clonal analysis. Colonies were analysed for the presence of Schwann cells, melanocytes, adrenergic cells or sensory-like cells. We provide for the first time a description of the temporal pattern of lineage segregation in neural crest cultures. In the absence of exogenous EDN3, crest cells proliferate and then differentiate. Colony assay indicates that in these differentiated cultures few undifferentiated precursors remain and there is a low replating efficiency. By contrast, in the presence of 100 ng/ml EDN3 differentiation is inhibited and most of the cells maintain the ability to give rise to mixed colonies and clones containing neural crest derivatives. A high replating efficiency is maintained. In secondary culture there was a progressive decline in the number of cell types per colony in control medium. This loss of developmental potential was not seen when exogenous EDN3 was present. Cell type analysis suggests two novel cellular targets for EDN3 under these conditions. Contrary to expectations, one is a multipotent precursor whose descendants include melanocytes, adrenergic cells and sensory-like cells; the other can give rise to melanocytes and Schwann cells. Our data do not support previous claims that the action of EDN3 in neural crest culture is selective for cells in the melanocyte lineage.

In vitro and in vivo differentiation of boundary cap neural crest stem cells into mature Schwann cells

2006 ◽  
Vol 198 (2) ◽  
pp. 438-449 ◽  
Author(s):  
Jorge B. Aquino ◽  
Jens Hjerling-Leffler ◽  
Martin Koltzenburg ◽  
Thomas Edlund ◽  
Marcelo J. Villar ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neural Crest ◽  
Schwann Cells ◽  
Neural Crest Stem Cells ◽  
In Vivo Differentiation

scholarly journals An in vitro ovarian explant culture system to examine sex change in a hermaphroditic fish

2020 ◽  
Author(s):  
Alexander Goikoetxea ◽  
Erin L Damsteegt ◽  
Erica V Todd ◽  
Andrew McNaughton ◽  
Neil J Gemmell ◽  
...  
Keyword(s):  
Culture System ◽  
Sex Change ◽  
Explant Culture ◽  
Culture Technique ◽  
Aromatase Activity ◽  
Histological Images ◽  
17Β Estradiol ◽  
Learning Software

AbstractMany teleost fishes undergo natural sex change, and elucidating the physiological and molecular controls of this process offers unique opportunities not only to develop methods of controlling sex in aquaculture settings, but to better understand vertebrate sexual development more broadly. Induction of sex change in some sequentially hermaphroditic or gonochoristic fish can be achieved in vivo through social manipulation, inhibition of aromatase activity, and steroid treatment. However, the induction of sex change in vitro has been largely unexplored. In this study, we established an in vitro culture system for ovarian explants in serum-free medium for a model sequential hermaphrodite, the New Zealand spotty wrasse (Notolabrus celidotus). This culture technique enabled evaluating the effect of various treatments with 17β-estradiol (E2), 11-ketotestosterone (11KT) or cortisol (CORT) on spotty wrasse ovarian architecture for 21 days. A quantitative approach to measuring the degree of ovarian atresia within histological images was also developed, using pixel-based machine learning software. Ovarian atresia likely due to culture was observed across all treatments including no-hormone controls, but was minimised with treatment of at least 10 ng/mL E2. Neither 11KT nor CORT administration induced proliferation of spermatogonia (i.e. sex change) in the cultured ovaries indicating culture beyond 21 days may be needed to induce sex change in vitro. The in vitro gonadal culture and analysis systems established here enable future studies investigating the paracrine role of sex steroids, glucocorticoids and a variety of other factors during gonadal sex change in fish.

Xanthine oxidase/dehydrogenase in mammary gland of mouse: relationship to mammogenesis and lactogenesis in vivo and in vitro

1991 ◽  
Vol 58 (4) ◽  
pp. 401-409 ◽  
Author(s):  
Thomas J. Hayden ◽  
Denise Brennan ◽  
Katherine Quirke ◽  
Paddie Murphy
Keyword(s):  
Mammary Gland ◽  
Xanthine Oxidase ◽  
Post Partum ◽  
Explant Culture ◽  
Placental Lactogen ◽  
Early Event ◽  
Histological Differentiation ◽  
Variant Enzyme

SummaryXanthine oxidase/dehydrogenase (XO/XDH) increases at mid gestation in mammary gland but not in liver of the mouse and remains elevated until the pups are weaned at 20 d post partum. The increase in enzyme activity is due neither to alteration in activators or inhibitors nor to a production of a variant enzyme with altered catalytic properties. The increase is preceded in vivo by a surge of prolactin-like activity (placental lactogen) in plasma, and prolactin is required for induction of XO/XDH in explant culture in vitro. Induction of XO/XDH in vivo and in vitro precedes the full histological differentiation of the gland. In addition, induction of XO/XDH in vitro occurs more rapidly and at lower concentrations of prolactin than does histological differentiation. Thus although XO/XDH is present in milk, increased XO/XDH activity is an early event in mammogenesis in vivo and in vitro rather than a terminal component of differentiation.

scholarly journals Lineage-specific requirements of β-catenin in neural crest development

2002 ◽  
Vol 159 (5) ◽  
pp. 867-880 ◽  
Author(s):  
Lisette Hari ◽  
Véronique Brault ◽  
Maurice Kléber ◽  
Hye-Youn Lee ◽  
Fabian Ille ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Neural Crest ◽  
Neural Crest Cells ◽  
Neural Crest Stem Cells ◽  
Downstream Signaling ◽  
Neural Crest Development ◽  
Sensory Neurogenesis

β-Catenin plays a pivotal role in cadherin-mediated cell adhesion. Moreover, it is a downstream signaling component of Wnt that controls multiple developmental processes such as cell proliferation, apoptosis, and fate decisions. To study the role of β-catenin in neural crest development, we used the Cre/loxP system to ablate β-catenin specifically in neural crest stem cells. Although several neural crest–derived structures develop normally, mutant animals lack melanocytes and dorsal root ganglia (DRG). In vivo and in vitro analyses revealed that mutant neural crest cells emigrate but fail to generate an early wave of sensory neurogenesis that is normally marked by the transcription factor neurogenin (ngn) 2. This indicates a role of β-catenin in premigratory or early migratory neural crest and points to heterogeneity of neural crest cells at the earliest stages of crest development. In addition, migratory neural crest cells lateral to the neural tube do not aggregate to form DRG and are unable to produce a later wave of sensory neurogenesis usually marked by the transcription factor ngn1. We propose that the requirement of β-catenin for the specification of melanocytes and sensory neuronal lineages reflects roles of β-catenin both in Wnt signaling and in mediating cell–cell interactions.

Further studies on the melanophores of periodic albino mutant of Xenopus laevis

Development ◽  
1986 ◽  
Vol 91 (1) ◽  
pp. 65-78
Author(s):  
T. Fukuzawa ◽  
H. Ide
Keyword(s):  
Xenopus Laevis ◽  
Microscopic Level ◽  
Wild Type ◽  
Light Microscopic Level ◽  
In Vitro Proliferation ◽  
Albino Mutant ◽  
Site Of Action ◽  
Periodic Albino

It is still unknown why dermal melanophores disappear during larval development, and why no or very few epidermal melanophores appear during and after metamorphosis, in Xenopus laevis showing periodic albinism (ap). To elucidate these points, we investigated (1) the occurrence of depigmentation in mutant (ap/ap) melanophores during in vitro proliferation and (2) the incidence of melanophore differentiation from mutant melanoblasts in the skin in vitro. During in vitro proliferation of mutant melanophores, ap-type melanosomes decreased in number gradually and instead the number of premelanosomes increased in the cells, which caused depigmentation at the light microscopic level in the culture. Depigmentation was observed only in mutant melanophores, and not in wild-type (+/+) melanophores. These results suggest that autonomous depigmentation of mutant dermal melanophores is the cause of the disappearance of these cells in vivo. Dopa-positive melanoblasts were demonstrated in both wild-type and mutant skins. However, the melanoblasts of metamorphosed mutant froglets did not differentiate in vitro, while those of wild-type froglets did. These results suggest that mutant melanoblasts in the skin of froglets lose the potency to differentiate into melanophores, and that this causes the lack of mutant melanophores in the froglets. The site of action of the ap gene is also discussed.

Taste buds develop autonomously from endoderm without induction by cephalic neural crest or paraxial mesoderm

Development ◽  
1997 ◽  
Vol 124 (5) ◽  
pp. 949-957 ◽  
Author(s):  
L.A. Barlow ◽  
R.G. Northcutt
Keyword(s):  
Epithelial Cells ◽  
Neural Crest ◽  
Explant Culture ◽  
Nerve Fibers ◽  
Taste Buds ◽  
Paraxial Mesoderm ◽  
Culture Techniques ◽  
Well Differentiated ◽  
Neural Crest Migration

Although it had long been believed that embryonic taste buds in vertebrates were induced to differentiate by ingrowing nerve fibers, we and others have recently shown that embryonic taste buds can develop normally in the complete absence of innervation. This leads to the question of which tissues, if any, induce the formation of taste buds in oropharyngeal endoderm. We proposed that taste buds, like many specialized epithelial cells, might arise via an inductive interaction between the endodermal epithelial cells that line the oropharynx and the adjacent mesenchyme that is derived from both cephalic neural crest and paraxial mesoderm. Using complementary grafting and explant culture techniques, however, we have now found that well-differentiated taste buds will develop in tissue completely devoid of neural crest and paraxial mesoderm derivatives. When the presumptive oropharyngeal region was removed from salamander embryos prior to the onset of cephalic neural crest migration, taste buds developed in grafts and explants coincident with their appearance in intact control embryos. Similarly, explants from neurulae in which movement of paraxial mesoderm had not yet begun also developed taste buds after 9–12 days in vitro. We conclude that neither cranial neural crest nor paraxial mesoderm is responsible for the induction of embryonic taste buds. Surprisingly, the ability to develop taste buds late in embryonic development seems to be an intrinsic feature of the oropharyngeal endoderm that is determined by the completion of gastrulation.

Sign in / Sign up

Export Citation Format

Share Document