Exo utero Surgery

Ngô-Muller, Valérie; Muneoka, Ken

doi:10.1385/1-59259-685-1:481

Cited by 8 publications

(8 citation statements)

References 18 publications

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“…For in vivo cell marking, injection of DiI (CellTracker TM CM-DiI, Invitrogen; 0.1 mg/ml in 0.3 M sucrose,1% ethanol, and 0.05% Nile blue sulfate,) was performed using pulled glass micropipettes and a microinjector as previously described [26], [27]. The DiI solution was injected into the dorsal connective tissue between the nail and the P3 element at 3 days post-amputation (DPA) to minimize non-specific labeling associated with enhanced cell necrosis following amputation injury.…”

Section: Methodsmentioning

confidence: 99%

Connective Tissue Fibroblast Properties Are Position-Dependent during Mouse Digit Tip Regeneration

Wang

Karapetyan

et al. 2013

PLoS ONE

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A key factor that contributes to the regenerative ability of regeneration-competent animals such as the salamander is their use of innate positional cues that guide the regeneration process. The limbs of mammals has severe regenerative limitations, however the distal most portion of the terminal phalange is regeneration competent. This regenerative ability of the adult mouse digit is level dependent: amputation through the distal half of the terminal phalanx (P3) leads to successful regeneration, whereas amputation through a more proximal location, e.g. the subterminal phalangeal element (P2), fails to regenerate. Do the connective tissue cells of the mammalian digit play a role similar to that of the salamander limb in controlling the regenerative response? To begin to address this question, we isolated and cultured cells of the connective tissue surrounding the phalangeal bones of regeneration competent (P3) and incompetent (P2) levels. Despite their close proximity and localization, these cells show very distinctive profiles when characterized in vitro and in vivo. In vitro studies comparing their proliferation and position-specific interactions reveal that cells isolated from the P3 and P2 are both capable of organizing and differentiating epithelial progenitors, but with different outcomes. The difference in interactions are further characterized with three-dimension cultures, in which P3 regenerative cells are shown to lack a contractile response that is seen in other fibroblast cultures, including the P2 cultures. In in vivo engraftment studies, the difference between these two cell lines is made more apparent. While both P2 and P3 cells participated in the regeneration of the terminal phalanx, their survival and proliferative indices were distinct, thus suggesting a key difference in their ability to interact within a regeneration permissive environment. These studies are the first to demonstrate distinct positional characteristics of connective tissue cells that are associated with their regenerative capabilities.

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Section: Methodsmentioning

confidence: 99%

Connective Tissue Fibroblast Properties Are Position-Dependent during Mouse Digit Tip Regeneration

Wang

Karapetyan

et al. 2013

PLoS ONE

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“…I. Saitou) were also used. Exo utero surgery was followed as described (Muneoka et al, 1986a,b;Ngo-Muller and Muneoka, 2000) with modifications.…”

Section: Methodsmentioning

confidence: 99%

Generation of Reelin-Positive Marginal Zone Cells from the Caudomedial Wall of Telencephalic Vesicles

Takiguchi-Hayashi¹,

Sekiguchi²,

Ashigaki³

et al. 2004

J. Neurosci.

220

190

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An early and fundamental step of the laminar organization of developing neocortex is controlled by the developmental programs that critically depend on the activities of reelin-positive cells in the marginal zone. However, the ontogeny of reelin-positive cells remained elusive. To gain insights into the spatial and temporal regulation of reelin-positive marginal zone cell development, we used a transgenic mouse line in which we defined the green fluorescent protein (GFP) transgene as a novel reliable molecular marker of reelin-positive marginal zone cells from the early stages of their development. We further used exo utero electroporation-mediated gene transfer that allows us to mark progenitor cells and monitor the descendants in the telencephalon in vivo. We show here the generation of reelinpositive marginal zone cells from the caudomedial wall of telencephalic vesicles, including the cortical hem, where the prominent expression of GFP is initially detected. These neurons tangentially migrate at the cortical marginal zone and are distributed throughout the entire neocortex in a caudomedial-high to rostrolateral-low gradient during the dynamic developmental period of corticogenesis. Therefore, our findings on reelin-positive marginal zone cells, in addition to the cortical interneurons, add to the emerging view that the neocortex consists of neuronal subtypes that originate from a focal source extrinsic to the neocortex, migrate tangentially into the neocortex, and thereby underlie neural organization of the neocortex.

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“…For in vivo studies, stage 11 fetal digit tips were amputated at E14.5 using exo utero surgical techniques as described (Reginelli et al, 1995;Ngo-Muller and Muneoka, 2000a). Briefly, timed-pregnant mice were anesthetized with sodium pentobarbital (60 µg/g body weight), fentany (1.6 µg/animal) and droperidol (80 µg/animal).…”

Section: Digit Amputationmentioning

confidence: 99%

Digit regeneration is regulated byMsx1andBMP4in fetal mice

et al. 2003

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The regeneration of digit tips in mammals, including humans and rodents,represents a model for organ regeneration in higher vertebrates. We had previously characterized digit tip regeneration during fetal and neonatal stages of digit formation in the mouse and found that regenerative capability correlated with the expression domain of the Msx1 gene. Using the stage 11 (E14.5) digit, we now show that digit tip regeneration occurs in organ culture and that Msx1, but not Msx2, mutant mice display a regeneration defect. Associated with this phenotype, we find that Bmp4 expression is downregulated in the Msx1 mutant digit and that mutant digit regeneration can be rescued in a dose-dependent manner by treatment with exogenous BMP4. Studies with the BMP-binding protein noggin show that wild-type digit regeneration is inhibited without inhibiting the expression of Msx1, Msx2 or Bmp4. These data identify a signaling pathway essential for digit regeneration, in which Msx1functions to regulate BMP4 production. We also provide evidence that endogenous Bmp4 expression is regulated by the combined activity of Msx1 and Msx2 in the forming digit tip; however, we discovered a compensatory Msx2 response that involves an expansion into the wild-type Msx1 domain. Thus, although both Msx1 and Msx2 function to regulate Bmp4 expression in the digit tip,the data are not consistent with a model in which Msx1 and Msx2 serve completely redundant functions in the regeneration response. These studies provide the first functional analysis of mammalian fetal digit regeneration and identify a new function for Msx1 and BMP4 as regulators of the regenerative response.

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Exo utero Surgery

Cited by 8 publications

References 18 publications

Connective Tissue Fibroblast Properties Are Position-Dependent during Mouse Digit Tip Regeneration

Connective Tissue Fibroblast Properties Are Position-Dependent during Mouse Digit Tip Regeneration

Generation of Reelin-Positive Marginal Zone Cells from the Caudomedial Wall of Telencephalic Vesicles

Digit regeneration is regulated byMsx1andBMP4in fetal mice

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