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The snail family gene snai3 is not essential for embryogenesis in mice.

Bradley CK, Norton CR, Chen Y, Han X, Booth CJ, Yoon JK, Krebs LT, Gridley T - PLoS ONE (2013)

Bottom Line: Three members of the Snail gene family have been described in mammals, encoded by the Snai1, Snai2, and Snai3 genes.However, little is known about the function of the Snai3 gene and protein.We also generated an EYFP-tagged Snai3 allele that accurately reflects endogenous Snai3 gene expression, with the highest levels of expression detected in thymus and skeletal muscle.

View Article: PubMed Central - PubMed

Affiliation: Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, Maine, United States of America.

ABSTRACT
The Snail gene family encodes zinc finger-containing transcriptional repressor proteins. Three members of the Snail gene family have been described in mammals, encoded by the Snai1, Snai2, and Snai3 genes. The function of the Snai1 and Snai2 genes have been studied extensively during both vertebrate embryogenesis and tumor progression and metastasis, and play critically important roles during these processes. However, little is known about the function of the Snai3 gene and protein. We describe here generation and analysis of Snai3 conditional and mutant mice. We also generated an EYFP-tagged Snai3 allele that accurately reflects endogenous Snai3 gene expression, with the highest levels of expression detected in thymus and skeletal muscle. Snai3 mutant homozygous mice are viable and fertile, and exhibit no obvious phenotypic defects. These results demonstrate that Snai3 gene function is not essential for embryogenesis in mice.

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Expression of the Snai3-EYFP allele.(A) EYFP expression is observed in thymus and in leg skeletal muscle at E15.5, and in heart, first branchial arch and somites at E10.5. EYFP expression was visualized by fluorescent stereomicroscopy. Dotted lines in the left panel outline the wild type thymus lobes. (B) Flow cytometry analysis demonstrated EYFP expression in Pre-B and B cells, but not in Pro-B cells of adult mouse bone marrow. (C) Flow cytometry analysis of thymus demonstrated EYFP expression in most T cell subsets. EYFP expression was highest in CD8 single positive T cells. (D) Quantitative RT-PCR of endogenous Snai3 RNA expression in the indicated organs of three (red bars) and seven (blue bars) week old wild type mice. Expression was normalized to βeta actin RNA levels in each organ.
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pone-0065344-g002: Expression of the Snai3-EYFP allele.(A) EYFP expression is observed in thymus and in leg skeletal muscle at E15.5, and in heart, first branchial arch and somites at E10.5. EYFP expression was visualized by fluorescent stereomicroscopy. Dotted lines in the left panel outline the wild type thymus lobes. (B) Flow cytometry analysis demonstrated EYFP expression in Pre-B and B cells, but not in Pro-B cells of adult mouse bone marrow. (C) Flow cytometry analysis of thymus demonstrated EYFP expression in most T cell subsets. EYFP expression was highest in CD8 single positive T cells. (D) Quantitative RT-PCR of endogenous Snai3 RNA expression in the indicated organs of three (red bars) and seven (blue bars) week old wild type mice. Expression was normalized to βeta actin RNA levels in each organ.

Mentions: We assessed expression of the Snai3-EYFP allele to determine how closely it matched Snai3 RNA expression. The sites of highest Snai3 RNA expression are skeletal muscle and thymus [6], [9]. By fluorescent stereomicroscopy, Snai3-EYFP expression was readily detected in both embryonic thymus and skeletal muscle at E15.5 (Fig. 2A). At E10.5, expression also was detected in both heart and the maxillary and mandibular portions of the first branchial arch (Fig. 2A). Both these sites of expression have been reported previously [6], [10]. We used flow cytometry to examine in bone marrow cells and thymocytes expression of both the Snai3-EYFP allele and lineage markers of lymphoid cells. As expected [8], [9], Snai3-EYFP expression was evident in both T and B cell populations (Fig. 2B, C). In the bone marrow, greater than 90% of EYFP-positive cells also expressed the pan B cell specific marker B220 (Fig. 2B). Notably, the percentage of EYFP-positive cells was greater in more mature B cell populations. Within the thymus, EYFP-positive cells were evident in all subpopulations during T cell development (Fig. 2C). However, the percentage of EYFP-positive cells was greater in very early T cells (DN1), as well as mature CD8 single positive T cells (80%). All sites of Snai3-EYFP expression were also sites of Snai3 RNA expression, as detected by quantitative RT-PCR of organs from adult mice (Fig. 2D), demonstrating that the Snai3-EYFP allele is a useful and accurate reporter of Snai3 gene expression.


The snail family gene snai3 is not essential for embryogenesis in mice.

Bradley CK, Norton CR, Chen Y, Han X, Booth CJ, Yoon JK, Krebs LT, Gridley T - PLoS ONE (2013)

Expression of the Snai3-EYFP allele.(A) EYFP expression is observed in thymus and in leg skeletal muscle at E15.5, and in heart, first branchial arch and somites at E10.5. EYFP expression was visualized by fluorescent stereomicroscopy. Dotted lines in the left panel outline the wild type thymus lobes. (B) Flow cytometry analysis demonstrated EYFP expression in Pre-B and B cells, but not in Pro-B cells of adult mouse bone marrow. (C) Flow cytometry analysis of thymus demonstrated EYFP expression in most T cell subsets. EYFP expression was highest in CD8 single positive T cells. (D) Quantitative RT-PCR of endogenous Snai3 RNA expression in the indicated organs of three (red bars) and seven (blue bars) week old wild type mice. Expression was normalized to βeta actin RNA levels in each organ.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3675094&req=5

pone-0065344-g002: Expression of the Snai3-EYFP allele.(A) EYFP expression is observed in thymus and in leg skeletal muscle at E15.5, and in heart, first branchial arch and somites at E10.5. EYFP expression was visualized by fluorescent stereomicroscopy. Dotted lines in the left panel outline the wild type thymus lobes. (B) Flow cytometry analysis demonstrated EYFP expression in Pre-B and B cells, but not in Pro-B cells of adult mouse bone marrow. (C) Flow cytometry analysis of thymus demonstrated EYFP expression in most T cell subsets. EYFP expression was highest in CD8 single positive T cells. (D) Quantitative RT-PCR of endogenous Snai3 RNA expression in the indicated organs of three (red bars) and seven (blue bars) week old wild type mice. Expression was normalized to βeta actin RNA levels in each organ.
Mentions: We assessed expression of the Snai3-EYFP allele to determine how closely it matched Snai3 RNA expression. The sites of highest Snai3 RNA expression are skeletal muscle and thymus [6], [9]. By fluorescent stereomicroscopy, Snai3-EYFP expression was readily detected in both embryonic thymus and skeletal muscle at E15.5 (Fig. 2A). At E10.5, expression also was detected in both heart and the maxillary and mandibular portions of the first branchial arch (Fig. 2A). Both these sites of expression have been reported previously [6], [10]. We used flow cytometry to examine in bone marrow cells and thymocytes expression of both the Snai3-EYFP allele and lineage markers of lymphoid cells. As expected [8], [9], Snai3-EYFP expression was evident in both T and B cell populations (Fig. 2B, C). In the bone marrow, greater than 90% of EYFP-positive cells also expressed the pan B cell specific marker B220 (Fig. 2B). Notably, the percentage of EYFP-positive cells was greater in more mature B cell populations. Within the thymus, EYFP-positive cells were evident in all subpopulations during T cell development (Fig. 2C). However, the percentage of EYFP-positive cells was greater in very early T cells (DN1), as well as mature CD8 single positive T cells (80%). All sites of Snai3-EYFP expression were also sites of Snai3 RNA expression, as detected by quantitative RT-PCR of organs from adult mice (Fig. 2D), demonstrating that the Snai3-EYFP allele is a useful and accurate reporter of Snai3 gene expression.

Bottom Line: Three members of the Snail gene family have been described in mammals, encoded by the Snai1, Snai2, and Snai3 genes.However, little is known about the function of the Snai3 gene and protein.We also generated an EYFP-tagged Snai3 allele that accurately reflects endogenous Snai3 gene expression, with the highest levels of expression detected in thymus and skeletal muscle.

View Article: PubMed Central - PubMed

Affiliation: Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, Maine, United States of America.

ABSTRACT
The Snail gene family encodes zinc finger-containing transcriptional repressor proteins. Three members of the Snail gene family have been described in mammals, encoded by the Snai1, Snai2, and Snai3 genes. The function of the Snai1 and Snai2 genes have been studied extensively during both vertebrate embryogenesis and tumor progression and metastasis, and play critically important roles during these processes. However, little is known about the function of the Snai3 gene and protein. We describe here generation and analysis of Snai3 conditional and mutant mice. We also generated an EYFP-tagged Snai3 allele that accurately reflects endogenous Snai3 gene expression, with the highest levels of expression detected in thymus and skeletal muscle. Snai3 mutant homozygous mice are viable and fertile, and exhibit no obvious phenotypic defects. These results demonstrate that Snai3 gene function is not essential for embryogenesis in mice.

Show MeSH
Related in: MedlinePlus