scholarly journals SCA-1/Ly6A Mesodermal Skeletal Progenitor Subpopulations Reveal Differential Commitment of Early Limb Bud Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Jessica Cristina Marín-Llera ◽  
Carlos Ignacio Lorda-Diez ◽  
Juan Mario Hurle ◽  
Jesús Chimal-Monroy
Keyword(s):  
Cell Fate ◽  
Limb Development ◽  
Developmental Stages ◽  
Limb Bud ◽  
Its Gene ◽  
Undifferentiated Cells ◽  
Advanced Stages ◽  
Cell Subpopulations ◽  
Inductive Signals

At early developmental stages, limb bud mesodermal undifferentiated cells are morphologically indistinguishable. Although the identification of several mesodermal skeletal progenitor cell populations has been recognized, in advanced stages of limb development here we identified and characterized the differentiation hierarchy of two new early limb bud subpopulations of skeletal progenitors defined by the differential expression of the SCA-1 marker. Based on tissue localization of the mesenchymal stromal cell-associated markers (MSC-am) CD29, Sca-1, CD44, CD105, CD90, and CD73, we identified, by multiparametric analysis, the presence of cell subpopulations in the limb bud capable of responding to inductive signals differentially, namely, sSca+ and sSca– cells. In concordance with its gene expression profile, cell cultures of the sSca+ subpopulation showed higher osteogenic but lower chondrogenic capacity than those of sSca–. Interestingly, under high-density conditions, fibroblast-like cells in the sSca+ subpopulation were abundant. Gain-of-function employing micromass cultures and the recombinant limb assay showed that SCA-1 expression promoted tenogenic differentiation, whereas chondrogenesis is delayed. This model represents a system to determine cell differentiation and morphogenesis of different cell subpopulations in similar conditions like in vivo. Our results suggest that the limb bud is composed of a heterogeneous population of progenitors that respond differently to local differentiation inductive signals in the early stages of development, where SCA-1 expression may play a permissive role during cell fate.

The application of aqueous two-phase partition to the study of chick limb mesenchymal diversification

Development ◽  
1986 ◽  
Vol 94 (1) ◽  
pp. 267-275
Author(s):  
C. P. Cottrill ◽  
Paul T. Sharpe ◽  
Lewis Wolpert
Keyword(s):  
Pattern Formation ◽  
Limb Development ◽  
Developmental Stages ◽  
Proximal Region ◽  
Limb Bud ◽  
Cell Populations ◽  
Two Phase ◽  
Phase Partition ◽  
Surface Character ◽  
Two Phase Partition

A technique which identifies cells differing in surface character, aqueous two-phase partition using thin-layer countercurrent distribution (TLCCD), has been used to study differentiation and pattern formation in the developing chick limb bud. The TLCCD profiles of cell populations, derived from various regions of morphologically undifferentiated mesenchyme from three different stages of limb development, have been compared. At no stage, or location, has the population been found to be homogeneous. Cells from progress zones and more proximal regions could all be resolved into several populations. The populations from progress zones at three different developmental stages were qualitatively similar but differed in the proportions of cells in each. The most striking differences in cell populations were those obtained from the most proximal region of the limb, closest to the flank, which represents the developmentally most advanced region.

scholarly journals The spatiotemporal expression patterns of MSC-associated markers contribute to the identification of progenitor subpopulations in developing limbs

2020 ◽  
Vol 64 (10-11-12) ◽  
pp. 499-506
Author(s):  
Argelia S. García-Cervera ◽  
Jesús Chimal-Monroy ◽  
Jessica C. Marín-llera
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Limb Development ◽  
Expression Patterns ◽  
Spatiotemporal Expression ◽  
Undifferentiated Cells ◽  
Adult Mscs

During limb development, skeletal tissues differentiate from their progenitor cells in an orchestrated manner. Mesenchymal stromal cells (MSCs), which are considered to be adult undifferentiated/progenitor cells, have traditionally been identified by the expression of MSC-associated markers (MSC-am) and their differentiation capacities. However, although MSCs have been isolated from bone marrow and a variety of adult tissues, their developmental origin is poorly understood. Remarkably, adult MSCs share similar differentiation characteristics with limb progenitors. Here, we determined the expression patterns of common MSC-am throughout mouse hindlimb development. Our results demonstrate that MSC-am expression is not restricted to undifferentiated cells in vivo. Results from the analysis of MSC-am spatiotemporal expression in the embryonic hindlimb allowed us to propose five subpopulations which represent all limb tissues that potentially correspond to progenitor cells for each lineage. This work contributes to the understanding of MSC-am expression dynamics throughout development and underlines the importance of considering their expression patterns in future MSC studies of the limb.

Limb development in mouse embryos: protection against teratogenic effects of 6-diazo-5 oxo-L-norleucine (DON) in vivo and in vitro

Development ◽  
1975 ◽  
Vol 33 (2) ◽  
pp. 355-370
Author(s):  
R. M. Greene ◽  
D. M. Kochhar
Keyword(s):  
Limb Development ◽  
Cleft Lip ◽  
Phenotypic Expression ◽  
Limb Bud ◽  
Concurrent Administration ◽  
Median Cleft ◽  
Mouse Limb ◽  
Limb Malformations

The glutamine analogue, 6-diazo-5-oxo-L-norleucine (DON), has been shown to inhibit biosynthesis of purines and glycosaminoglycans, presumably by blocking the glutaminedependent steps in the biosynthetic pathways. The teratogenic potential of DON on the developing mouse limb-bud in vivo and in vitro was studied in an attempt to discriminate whether DON is exerting its teratogenic effect by interfering with glycosaminoglycan orpurine metabolism. A single intramuscular injection of DON (0·5 mg/kg) to ICR/DUB mice on day 10 of gestation resulted in 76% resorption, while fetuses surviving to day 17 exhibited growth retardation, median cleft lip, and limb malformations. Concurrent administration of Lglutamine (250 mg/kg) provided no protection against resorption or malformations, while 5-aminoimidazolecarboxamide (AIC, 250 mg/kg) decreased the resorption rate to 34% without significantly altering the incidence of malformations. Injection of DON alone on day 11 resulted in 87% of fetuses exhibiting limb malformations, with only 2% resorption. Concurrent injection of AIC decreased the frequency of limb malformations to 32%. L-Glutamine, D-glucosamine, or inosinic acid were without any protective effect in vivo. DON (5 μg/ml medium) added in vitro to organ cultures of day 11 mouse limb-buds caused all limbs to evidence cartilage abnormalities. In this system, either L-glutamine or D-glucosamine (0·5 mg/ml medium) provided protection against DON effects while AIC (0·5 mg/ml medium) offered no protection in vitro. These data suggest that DON exerts its effects in vivo by interfering with purine metabolism while in vitro its teratogenic action may be interruption of glycosaminoglycan biosynthesis. This may reflect upon the relative importance of growth and differentiation to limb development in vivo and in vitro. These data infer that limb development in vitro relies more on the differentiative process (differentiation of cartilage) than on growth, whereas limb development in vivo is dependent, at this stage, to a greater extent on growth for normal phenotypic expression.

scholarly journals The expression of slow myosin during mammalian somitogenesis and limb bud differentiation.

1988 ◽  
Vol 107 (6) ◽  
pp. 2191-2197 ◽  
Author(s):  
E Vivarelli ◽  
W E Brown ◽  
R G Whalen ◽  
G Cossu
Keyword(s):  
Monoclonal Antibodies ◽  
Developmental Stages ◽  
Mouse Embryos ◽  
Fiber Formation ◽  
Limb Bud ◽  
Slow Myosin ◽  
Fast Myosin ◽  
The Relationship ◽  
Cells Cultured

The developmental pattern of slow myosin expression has been studied in mouse embryos from the somitic stage to the period of secondary fiber formation and in myogenic cells, cultured from the same developmental stages. The results obtained, using a combination of different polyclonal and monoclonal antibodies, indicate that slow myosin is coexpressed in virtually all the cells that express embryonic (fast) myosin in somites and limb buds in vivo as well as in culture. On the contrary fetal or late myoblasts (from 15-d-old embryos) express in culture only embryonic (fast) myosin. At this stage, muscle cells in vivo, as already shown (Crow, M.T., and F.A. Stockdale. 1986. Dev. Biol. 113:238-254; Dhoot, G.K. 1986. Muscle & Nerve. 9:155-164; Draeger, A., A.G. Weeds, and R.B. Fitzsimons. 1987. J. Neurol. Sci. 81:19-43; Miller, J.B., and F.A. Stockdale. 1986. J. Cell Biol. 103:2197-2208), consist of primary myotubes, which express both myosins, and secondary myotubes, which express preferentially embryonic (fast) myosin. Under no circumstance neonatal or adult fast myosins were detected. Western blot analysis confirmed the immunocytochemical data. These results suggest that embryonic myoblasts in mammals are all committed to the mixed embryonic-(fast) slow lineage and, accordingly, all primary fibers express both myosins, whereas fetal myoblasts mostly belong to the embryonic (fast) lineage and likely generate fibers containing only embryonic (fast) myosin. The relationship with current models of avian myogenesis are discussed.

Normal limb development in conditional mutants of Fgf4

Development ◽  
2000 ◽  
Vol 127 (5) ◽  
pp. 989-996 ◽  
Author(s):  
A.M. Moon ◽  
A.M. Boulet ◽  
M.R. Capecchi
Keyword(s):  
Limb Development ◽  
Expression Patterns ◽  
Large Body ◽  
Embryonic Lethality ◽  
Limb Bud ◽  
Recombination Reaction ◽  
Site Specific ◽  
Early Embryonic Lethality ◽  
Limb Outgrowth

Fibroblast growth factors (FGFs) mediate multiple developmental signals in vertebrates. Several of these factors are expressed in limb bud structures that direct patterning of the limb. FGF4 is produced in the apical ectodermal ridge (AER) where it is hypothesized to provide mitogenic and morphogenic signals to the underlying mesenchyme that regulate normal limb development. Mutation of this gene in the germline of mice results in early embryonic lethality, preventing subsequent evaluation of Fgf4 function in the AER. A conditional mutant of Fgf4, based on site-specific Cre/loxP-mediated excision of the gene, allowed us to bypass embryonic lethality and directly test the role of FGF4 during limb development in living murine embryos. This conditional mutation was designed so that concomitant with inactivation of the Fgf4 gene by excision of all Fgf4-coding sequences, a reporter gene was activated in Fgf4-expressing cells, allowing assessment of the site-specific recombination reaction. Although a large body of evidence led us to predict that ablation of Fgf4 gene function in the AER of developing mice would result in abnormal limb outgrowth and patterning, we found that Fgf4 conditional mutants had normal limbs. Furthermore, expression patterns of Shh, Bmp2, Fgf8 and Fgf10 were normal in the limb buds of the conditional mutants. These findings indicate that the previously proposed FGF4-SHH feedback loop is not essential for coordination of murine limb outgrowth and patterning. We suggest that some of the roles currently attributed to FGF4 during early vertebrate limb development may be performed by other AER factors in vivo.

scholarly journals OCT4 impedes cell fate redirection by the melanocyte lineage master regulator MITF

2017 ◽  
Author(s):  
Danna Sheinboim ◽  
Itay Maza ◽  
Iris Dror ◽  
Shivang Parikh ◽  
Vladislav Krupalnik ◽  
...  
Keyword(s):  
Cell Fate ◽  
Transcriptional Activity ◽  
Developmental Stages ◽  
Ectopic Expression ◽  
Embryonic Stem ◽  
Master Regulator ◽  
Master Regulators ◽  
Differentiated Cells ◽  
Cell Fate Conversion

ABSTRACTEctopic expression of lineage master regulators induces transdifferentiation. Whether cell fate transitions can be induced during various developmental stages has never been systemically examined. Here we discovered that amongst different developmental stages, embryonic stem cells (ESCs) were resistant to cell fate conversion induced by the melanocyte lineage master regulator MITF. We generated a transgenic system and found that in ESCs, the pluripotency master regulator, OCT4, counteracts pro-differentiation induced by MITF by physical interference with MITF transcriptional activity. We further found that ESCs must be released from OCT4-maintained pluripotency prior to ectopically induced differentiation. Moreover, OCT4 induction in various differentiated cells repressed their lineage identity in vivo. Alongside, chromatin architecture combined with ChIP-seq analysis suggested that OCT4 competes with various lineage master regulators for binding promoters and enhancers. Our analysis reveals pluripotency and transdifferentiation regulatory principles and could open new opportunities in the field of regenerative medicine.

scholarly journals Changes in segmentation and setation along the anterior/posterior axis of the homonomous trunk limbs of a remipede (Crustacea, Arthropoda)

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2305 ◽  
Author(s):  
Viacheslav N. Ivanenko ◽  
Ekaterina A. Antonenko ◽  
Mikhail S. Gelfand ◽  
Jill Yager ◽  
Frank D. Ferrari
Keyword(s):  
Limb Development ◽  
Developmental Stages ◽  
The Body ◽  
Limb Bud ◽  
Vector Analysis ◽  
Contralateral Limb ◽  
Stage Of Development ◽  
Trunk Limbs ◽  
Anchialine Caves ◽  
Trunk Limb

This study describes the segmentation and setation at different developmental stages of the homonomous trunk limbs of the remipedeSpeleonectes tulumensisYager, 1987 collected in anchialine caves of the Yucatan Peninsula. Most homonomous trunk limbs originate ventrolaterally and are composed of two protopodal segments, three exopodal segments and four endopodal segments; contralateral limb pairs are united by a sternal bar. However, the last few posterior limbs originate ventrally, are smaller sized, and have regressively fewer segments, suggesting that limb development passes through several intermediate steps beginning with a limb bud. A terminal stage of development is proposed for specimens on which the posterior somite bears a simple bilobate limb bud, and the adjacent somite bears a limb with a protopod comprised of a coxapod and basipod, and with three exopodal and four endopodal segments. On each trunk limb there are 20 serially homologous groups of setae, and the numbers of setae on different limbs usually varies. These groups of setae are arranged linearly and are identified based on the morphology of the setae and their position on the segments. The number of setae in these groups increases gradually from the anterior homonomous limb to a maximum between limbs 8–12; the number then decreases sharply on the more posterior limbs. Changes in the number of setae, which reach a maximum between trunk limbs 8–12, differ from changes in segmentation which vary only over the last few posterior trunk limbs. Following a vector analysis that identified a spatial pattern for these 20 groups of setae among the different homonomous limbs, the hypothesis was confirmed that the number of setae in any given group and any given limb is correlated with the group, with the position of the somite along the body axis, and with the number of somites present on the specimens. This is the first vector analysis used to analyze a pattern of developmental changes in serially homologs of an arthropod. Development of remipede limbs are compared and contrasted with similar copepod limbs. Architecture, particularly the sternal bar uniting contralateral limb pairs, proposed as homologous, and development of trunk limb segmentation of the remipede is generally similar to that of copepods, but the remipede limb differs in several ways including an additional endopodal segment, the proximal, that appears simultaneously with the protopod during development.

Transcription Factor IIAτIs Associated with Undifferentiated Cells and Its Gene Expression Is Repressed in Primary Neurons at the Chromatin Level In Vivo

2006 ◽  
Vol 15 (2) ◽  
pp. 175-190 ◽  
Author(s):  
Mariko L. Howe ◽  
Zareen F. Mehmud ◽  
Shamol Saha ◽  
Michael Buratovich ◽  
Erica A. Stutius ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Primary Neurons ◽  
Its Gene ◽  
Undifferentiated Cells
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