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Garland Publishing, Inc; p. One significant limitation in generating transgenic reporter genes is that the regulatory elements that control the gene transcription can be scattered over large regions, therefore making their identification quite difficult and time consuming, especially when they are not conserved among species. Gradually, the staining expanded rostro-caudally along almost the entire length of the spinal cord Fig. For genetic engineering of such animals, mice have long been the most frequently used experimental vertebrate model [ 1 ]. Merged images obtained GFP staining and retrograde labeling were superimposed. Prestige MC The control of cell number in the lumbar ventral horns during the development of Xenopus laevis tadpoles. No fluorescence was expressed in the hindbrain, suggesting that some of the regulatory elements of the X. Merged images of uts2b staining and retrograde labeling.

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    Lateral views of tadpoles with dorsal side up and rostral to the right. However, organisms such as zebrafish and Xenopus have recently emerged as alternative systems and are now being widely exploited to study human neurological diseases as well as to screen for potential therapeutics [ 2 — 5 ]. Merged image obtained when uts2b and ChAT stainings were superimposed. Up to now, functional studies on the uts2b promoter have never been carried out in Xenopus.

    However, the first fluorescence signal could be detected as early as 32, but only as small dots in the rostral spinal cord. Combined fluorescent in situ hybridization and immunofluorescence In situ hybridization was performed before immunohistochemistry as described above and revealed using FITC- or TAMRA-conjugated tyramide. Dorsal view, rostral to the right.

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    All electrophysiological signals were computer-stored using a digitizer interface Digidata ; Pclamp10 software; Molecular Devices and analyzed off-line using the Clampfit software Molecular Devices.

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    Hereafter, motoneurons labeled from tail at stage 50 or 55 will be referred to as axial motoneurons, motoneurons labeled from dorsal trunk at stage 60 as thoracic dorsal motoneurons and motoneurons labeled from hindlimb buds at stage 50 and leg muscles at stage 55 or 60 as appendicular motoneurons [ 48 — 50 ]. Typically, olig2 is expressed within the progenitor domain that gives rise both to motoneurons and oligodendrocytes [ 17 ].

    Biochem Biophys Res Commun Despite their great interest, all these genes have the disadvantage of also being expressed in other neurons than motoneurons, not only in the brain but also in the spinal cord.

    Gradually, the staining expanded rostro-caudally along almost the entire length of the spinal cord Fig.

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    Author information Article notes Copyright and License information Disclaimer. Methods Cell Sci Adv Drug Deliv Rev 69— Becker TS, Rinkwitz S Zebrafish as a genomics model for human neurological and polygenic disorders.

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    Garland Publishing, Inc; p. In Xenopustrials of BAC transgenesis have been reported in only two studies so far [ 3561 ].

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    Since we found that uts2b was constitutively active in cells that correspond to spinal motoneurons, uts2b promoter was a good candidate to use to drive the expression of GFP in transgenic Xenopus motoneurons.

    Skyrah Bai 4, views. Cedric Raoul, Academic Editor. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

    Pharmacol Rev in press. Xenopus is an excellent tetrapod model for studying normal and pathological motoneuron ontogeny due to its developmental morpho-physiological advantages.

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    Red arrowheads indicate uts2b— retrograde labeled cells. Adv Drug Deliv Rev 69— In late tadpole stage, uts2b mRNA was detected both in the hindbrain and in the spinal cord.

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    In the case of motoneurons, the regulatory sequences of several genes encoding transcription factors involved in motoneuron specification, such as HB9 also named Mnx1ISLET-1 ISL1 and OLIG2were shown to be particularly well suited, especially in zebrafish [ 13 — 15 ].

    In mice, it has been shown that a majority of spinal motoneurons simultaneously expresses uts2b and its paralog uts2 [ 27 ].

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    Crystals of fluorescent dextran amine dyes Invitrogen were applied intramuscularly with an insect pin.

    Support Center Support Center. Yet, such large DNA constructs can be replicated and then maintained as episomes over cell divisions.

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    Dorsal view of whole-mount preparation of hindbrain-spinal cord CNS. Dorsal view with rostral up. The first uts2b signal could be detected in the rostral spinal cord from stage 24, pre-hatching early tailbud embryos Fig.

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    1. Thus, the decrease in GFP expression observed during metamorphosis may not be due to any BAC loss but, rather, reflects the endogenous expression pattern of uts2b.

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    4. In Xenopustrials of BAC transgenesis have been reported in only two studies so far [ 3561 ].