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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 184 (1991), S. 153-157 
    ISSN: 1432-0568
    Keywords: Mouse embryos ; Limb bud ; Regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The ability of day E10 mouse fore-limb bud to regulate following the removal of a portion of limb tissue was investigated. A longitudinal strip of tissue, two to three somites in width and extending from the base of the limb bud to its distal tip, was excised. The embryos were then maintained in a roller culture system for periods of 6 h, 12 h or 24 h post-operatively prior to fixation and subsequent examination. The embryos were examined with scanning electron microscopy (SEM) and light microscopy. SEM revealed that about two thirds of the operated limbs grossly restored their overall morphology. The sequence of morphological changes involved in the restoration process is described. The ability of the restored limb bud to develop an apical ectodermal ridge (AER) is shown in histological sections.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 180 (1989), S. 293-300 
    ISSN: 1432-0568
    Keywords: Myogenic cell ; Limb bud ; Cell movement
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Quail limb mesenchyme containing myogenic cells of somitic origin were transplanted into chick limb buds to determine whether cell movement might play a role in avian limb myogenesis. In general, cell displacement was not detected 1-day after implantation: all quail cells were found at the graft site. Migration was evident 2-days after implantation but not all cell types were capable of movement; myogenic cells were very invasive while chondrocytes were relatively immobile. The spreading of myogenic cells was discernible up to 4-days after implantation and specifically in a proximodistal direction towards the apex of the limb.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 186 (1992), S. 67-74 
    ISSN: 1432-0568
    Keywords: Rat embryos ; Limb bud ; Regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The regulative potential of the fore-limb region following the removal of the limb bud was investigated in 11.5-day rat embryos. Fore-limb buds were amputated from a total of 54 embryos. Five embryos were immediately examined via scanning electron microscopy (SEM) to assess the quality of the operation and the reproducibility of the technique. In all cases, the forelimb bud and adjacent tissues extending down to the celomic cavity at the same level were completely removed. The remaining 49 operated embryos were cultured in vitro and examined at different time intervals. Gross inspection of embryos which had been cultured for 24 h, revealed that 24 out of 37 had developed a pair of limb bud-like protrusions at the operation site. Twelve formed only a single protrusion, while nothing was found in the remaining embryo. These protrusions were examined in greater detail under SEM and light microscopy. SEM observations showed that these protrusions were covered with an intact layer of ectoderm. In embryos with a pair of protrusions, these outgrowths developed opposite somites 7 to 13. The failure of either one of these two outgrowths to form, produced our second type of experimental embryo, those which had just a single protrusion. Histological examination revealed that an apical ectodermal ridge (AER) was discernible on the protrusions of 36% of the embryos. Finally, we have established how these protrusions were constructed from SEM observations of operated embryos cultured for 6 h and 10 h.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 184 (1991), S. 583-590 
    ISSN: 1432-0568
    Keywords: Avian embryos ; Latex beads ; Fibronectin ; Limb bud
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Latex beads were implanted into the chick wing bud to determine whether parameters other than active movement, for example matrix-driven translocation and growth of the limb bud, were responsible for the extensive re-allocation of myogenic cells that occurs during limb development. Latex beads were implanted into nine stage 20–24 Hamburger and Hamilton (H.H.) wing buds, and were allowed to develop for 3 days before examination. In all cases, it was found that most of the latex beads (86.57%±11.4%) were confined to the implantation site. A small percentage of beads was observed in the connective and myogenic regions proximal and distal to the graft side. In general the displacement of these beads was relatively short, although in one specimen a few beads were translocated to regions as far as the autopod. The surface of the latex beads was also coated with fibronectin prior to transplantation, to ascertain whether the extracellular matrix can influence the translocation of beads within the limb bud. Ten specimens were examined, and as for uncoated latex beads, most of the fibronectin-coated beads (87.14%±11.67%) were contained within the transplantation site. Again a mall percentage of beads was found in the connective and myogenic but not in the chondrogenic tissues proximal and distal to the graft side. In one specimen fibronectin-coated beads were translocated to regions in the autopod, but in general, bead displacement was relatively short. In sum, latex beads can not move to any great extent within the limb bud, and the coating of these beads with fibronectin did not influence bead translocation.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Developmental Dynamics 203 (1995), S. 324-336 
    ISSN: 1058-8388
    Keywords: Somites ; Limb bud ; Myogenic cells ; Transgenic embryos ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In this study, we have isolated newly formed somites from the caudal regions of 8.5 day mouse embryos and transplanted them orthotopically into correspondingly staged hosts at the level of the prospective limb-forming region. The experimental embryos were then cultured intact for 32-36 hr. The donor somites used were pre-labelled with DiI, a fluorescent lipophilic dye, or were obtained from transgenic embryos that carried a 1 kb 5′ regulatory sequence of the desmin gene linked to the gene encoding Escherichia coli β-galactosidase. The transgene is specifically expressed in skeletal muscles (Li et al. [1993] Development 117:947-959). The aim of these experiments was to show definitively that the musculature of the mammalian limb is derived from the somites. The results demonstrated that DiI-labelled cells from the implanted somites were able to invade the proximal region of the fore-limb bud during the course of development. The use of transgenic somites as grafts confirmed that some of the somitic cells found in the limbs were myogenic cells. To determine whether the displacement of somitic cells is an active or passive process, somatopleure obtained from the prospective limb-forming regions of day 8.5 day embryos was implanted into 8.5 day hosts. We did not detect the presence of DiI-labelled somatopleural cells in the fore-limb after 32-36 hr of culture. This suggests that somitic cells reached the limb bud via active locaomotion rather than as a result of being passively dragged there, as the limb elongates during development. In addition, we injected latex beads into the somites, as probes, to determine whether extracellular matrix-driven translocation plays a role in driving the somitic cells to the limb bud. In a majority of the specimens examined, we could not detect the presence of these beads in the limb bud. However, in the trunk of these embryos, the beads were found dispersed throughout the ventral neural crest pathway.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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