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BAC-Dkk3-EGFP transgenic mouse: an in vivo analytical tool for Dkk3 expression.

Muranishi Y, Furukawa T - J. Biomed. Biotechnol. (2012)

Bottom Line: Dickkopf (DKK) family proteins are secreted modulators of the Wnt signaling pathway and are capable of regulating the development of many organs and tissues.Expression analysis using the BAC-Dkk3-EGFP transgenic mice revealed that Dkk3 is expressed in retinal progenitor cells (RPCs) at embryonic stages and in Müller glial cells in the adult retina.Since Müller glial cells may play a potential role in retinal regeneration, BAC-Dkk3-EGFP mice could be useful for retinal regeneration studies.

View Article: PubMed Central - PubMed

Affiliation: CREST, Japan Science and Technology Agency, 3-2 Yamadaoka, Osaka, Suita, Japan.

ABSTRACT
Dickkopf (DKK) family proteins are secreted modulators of the Wnt signaling pathway and are capable of regulating the development of many organs and tissues. We previously identified Dkk3 to be a molecule predominantly expressed in the mouse embryonic retina. However, which cell expresses Dkk3 in the developing and mature mouse retina remains to be elucidated. To examine the precise expression of the Dkk3 protein, we generated BAC-Dkk3-EGFP transgenic mice that express EGFP integrated into the Dkk3 gene in a BAC plasmid. Expression analysis using the BAC-Dkk3-EGFP transgenic mice revealed that Dkk3 is expressed in retinal progenitor cells (RPCs) at embryonic stages and in Müller glial cells in the adult retina. Since Müller glial cells may play a potential role in retinal regeneration, BAC-Dkk3-EGFP mice could be useful for retinal regeneration studies.

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(a, b) Photographs of the BAC-DKK3-EGFP mouse embryo at E9.5. The embryo was visualized under bright-field (a) and fluorescence (b). h, heart; sc, spinal cord. (c) A coronal section through the palatal region of the BAC-Dkk3-EGFP mouse at E16.5. t, tooth; gm, genioglossus muscle. (d) A coronal section through the heart of the BAC-DKK3-EGFP mouse at E16.5. PT, pulmonary trunk; Ao, Aorta; LA, left atrium; RA, right atrium; RV, right ventricle. (e–j) EGFP expression in the BAC-Dkk3-EGFP mouse retina. EGFP expression was detected at E15.5 (e), E18.5 (f), P0 (g), P3 (h), P6 (i), and P14 (j). Nuclei were counterstained with DAPI. NBL, neuroblastic layer; ONL, outer nuclear layer; INL, inner nuclear layer. Scale bar represents 50 μm.
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fig2: (a, b) Photographs of the BAC-DKK3-EGFP mouse embryo at E9.5. The embryo was visualized under bright-field (a) and fluorescence (b). h, heart; sc, spinal cord. (c) A coronal section through the palatal region of the BAC-Dkk3-EGFP mouse at E16.5. t, tooth; gm, genioglossus muscle. (d) A coronal section through the heart of the BAC-DKK3-EGFP mouse at E16.5. PT, pulmonary trunk; Ao, Aorta; LA, left atrium; RA, right atrium; RV, right ventricle. (e–j) EGFP expression in the BAC-Dkk3-EGFP mouse retina. EGFP expression was detected at E15.5 (e), E18.5 (f), P0 (g), P3 (h), P6 (i), and P14 (j). Nuclei were counterstained with DAPI. NBL, neuroblastic layer; ONL, outer nuclear layer; INL, inner nuclear layer. Scale bar represents 50 μm.

Mentions: In the BAC-Dkk3-EGFP mouse embryo, we observed an EGFP expression in the eye, heart, and spinal cord at embryonic day 9.5 (E9.5) (Figures 2(a) and 2(b)). An EGFP signal was weakly detected in the palate and heart at E16.5 (Figures 2(c) and 2(d)). In our previous study, Dkk3 mRNA expression was detected in the entire NBL of the embryonic retina and in the INL in the postnatal retina using in situ hybridization [7]. To confirm this result, we investigated EGFP expression in the developing retina of the BAC-Dkk3-EGFP mouse. From E15.5 to postnatal day 0 (P0), EGFP was detected in most mitotic and undifferentiated RPCs throughout the NBL (Figures 2(e)–2(g)). At P3, EGFP expression was limited to RPCs, and differentiated retinal cells were not labeled by EGFP (Figure 2(h)). At P6, EGFP was expressed in Müller glia-like cells, which exhibit long glial fibers and localize their cell bodies in the INL (Figure 2(i)). At P14, only Müller glial cells in the INL-expressed EGFP, and we observed a large number of glial fibers extending into the surrounding retinal layers (Figure 2(j)).


BAC-Dkk3-EGFP transgenic mouse: an in vivo analytical tool for Dkk3 expression.

Muranishi Y, Furukawa T - J. Biomed. Biotechnol. (2012)

(a, b) Photographs of the BAC-DKK3-EGFP mouse embryo at E9.5. The embryo was visualized under bright-field (a) and fluorescence (b). h, heart; sc, spinal cord. (c) A coronal section through the palatal region of the BAC-Dkk3-EGFP mouse at E16.5. t, tooth; gm, genioglossus muscle. (d) A coronal section through the heart of the BAC-DKK3-EGFP mouse at E16.5. PT, pulmonary trunk; Ao, Aorta; LA, left atrium; RA, right atrium; RV, right ventricle. (e–j) EGFP expression in the BAC-Dkk3-EGFP mouse retina. EGFP expression was detected at E15.5 (e), E18.5 (f), P0 (g), P3 (h), P6 (i), and P14 (j). Nuclei were counterstained with DAPI. NBL, neuroblastic layer; ONL, outer nuclear layer; INL, inner nuclear layer. Scale bar represents 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3403811&req=5

fig2: (a, b) Photographs of the BAC-DKK3-EGFP mouse embryo at E9.5. The embryo was visualized under bright-field (a) and fluorescence (b). h, heart; sc, spinal cord. (c) A coronal section through the palatal region of the BAC-Dkk3-EGFP mouse at E16.5. t, tooth; gm, genioglossus muscle. (d) A coronal section through the heart of the BAC-DKK3-EGFP mouse at E16.5. PT, pulmonary trunk; Ao, Aorta; LA, left atrium; RA, right atrium; RV, right ventricle. (e–j) EGFP expression in the BAC-Dkk3-EGFP mouse retina. EGFP expression was detected at E15.5 (e), E18.5 (f), P0 (g), P3 (h), P6 (i), and P14 (j). Nuclei were counterstained with DAPI. NBL, neuroblastic layer; ONL, outer nuclear layer; INL, inner nuclear layer. Scale bar represents 50 μm.
Mentions: In the BAC-Dkk3-EGFP mouse embryo, we observed an EGFP expression in the eye, heart, and spinal cord at embryonic day 9.5 (E9.5) (Figures 2(a) and 2(b)). An EGFP signal was weakly detected in the palate and heart at E16.5 (Figures 2(c) and 2(d)). In our previous study, Dkk3 mRNA expression was detected in the entire NBL of the embryonic retina and in the INL in the postnatal retina using in situ hybridization [7]. To confirm this result, we investigated EGFP expression in the developing retina of the BAC-Dkk3-EGFP mouse. From E15.5 to postnatal day 0 (P0), EGFP was detected in most mitotic and undifferentiated RPCs throughout the NBL (Figures 2(e)–2(g)). At P3, EGFP expression was limited to RPCs, and differentiated retinal cells were not labeled by EGFP (Figure 2(h)). At P6, EGFP was expressed in Müller glia-like cells, which exhibit long glial fibers and localize their cell bodies in the INL (Figure 2(i)). At P14, only Müller glial cells in the INL-expressed EGFP, and we observed a large number of glial fibers extending into the surrounding retinal layers (Figure 2(j)).

Bottom Line: Dickkopf (DKK) family proteins are secreted modulators of the Wnt signaling pathway and are capable of regulating the development of many organs and tissues.Expression analysis using the BAC-Dkk3-EGFP transgenic mice revealed that Dkk3 is expressed in retinal progenitor cells (RPCs) at embryonic stages and in Müller glial cells in the adult retina.Since Müller glial cells may play a potential role in retinal regeneration, BAC-Dkk3-EGFP mice could be useful for retinal regeneration studies.

View Article: PubMed Central - PubMed

Affiliation: CREST, Japan Science and Technology Agency, 3-2 Yamadaoka, Osaka, Suita, Japan.

ABSTRACT
Dickkopf (DKK) family proteins are secreted modulators of the Wnt signaling pathway and are capable of regulating the development of many organs and tissues. We previously identified Dkk3 to be a molecule predominantly expressed in the mouse embryonic retina. However, which cell expresses Dkk3 in the developing and mature mouse retina remains to be elucidated. To examine the precise expression of the Dkk3 protein, we generated BAC-Dkk3-EGFP transgenic mice that express EGFP integrated into the Dkk3 gene in a BAC plasmid. Expression analysis using the BAC-Dkk3-EGFP transgenic mice revealed that Dkk3 is expressed in retinal progenitor cells (RPCs) at embryonic stages and in Müller glial cells in the adult retina. Since Müller glial cells may play a potential role in retinal regeneration, BAC-Dkk3-EGFP mice could be useful for retinal regeneration studies.

Show MeSH
Related in: MedlinePlus