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Myeloid ecotropic viral integration site 1 (MEIS) 1 involvement in embryonic implantation.

Xu B, Geerts D, Qian K, Zhang H, Zhu G - Hum. Reprod. (2008)

Bottom Line: Analysis of an Affymetrix data set in the public domain showed high expression of MEIS1 in human endometrium.MEIS1 expression was confirmed during the human menstrual cycle by RT-PCR and in situ hybridization and was increased during the secretory compared with proliferative phase of the cycle (P = 0.0001), the time of implantation.Increased or decreased Meis1 expression significantly increased or decreased the expression of integrin beta3, a transcriptional target of HOXA10 and an important factor in early embryo-endometrium interactions (P = 0.006).

View Article: PubMed Central - PubMed

Affiliation: Reproductive Medicine Center, Tongji Hospital, Tongji Medicine College, Huazhong, University of Science and Technology, 1095 JieFang Avenue, Wuhan 430030, People's Republic of China.

ABSTRACT

Background: The HOXA10 homeobox gene controls embryonic uterine development and adult endometrial receptivity. The three-amino-acid loop extension (TALE) family homeobox genes like myeloid ecotropic viral integration site 1 (MEIS) provide enhanced target gene activation and specificity in HOX-regulated cellular processes by acting as HOX cofactors.

Methods and results: Analysis of an Affymetrix data set in the public domain showed high expression of MEIS1 in human endometrium. MEIS1 expression was confirmed during the human menstrual cycle by RT-PCR and in situ hybridization and was increased during the secretory compared with proliferative phase of the cycle (P = 0.0001), the time of implantation. To assess the importance of maternal Meis1 expression in a mouse model, the uteri of Day 2 pregnant mice were injected with Meis1 over-expression or small interfering RNA (siRNA) constructs. Blocking Meis1 expression by siRNA before implantation significantly reduced average implantation rates (P = 0.00001). Increased or decreased Meis1 expression significantly increased or decreased the expression of integrin beta3, a transcriptional target of HOXA10 and an important factor in early embryo-endometrium interactions (P = 0.006). Manipulating Meis1 expression before implantation also dramatically affected the number of pinopodes, uterine endometrial epithelial projections that develop at the time of endometrial receptivity.

Conclusions: The results suggest that in mouse, meis1 contributes to regulating endometrial development during the menstrual cycle and establishing the conditions necessary for implantation.

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Effect of Meis1 expression manipulation on murine uterine integrin β3 (Itgb3) expression.(a) RT–PCR analysis of Itgb3 expression in murine endometria transfected with pcDNA4/MEIS1 (U), empty pcDNA4 (Uc), pTER/Meis1 siRNA (D) or control pTER constructs (Dc). A representative agarose EtBr picture is shown. No PCR product was observed in control samples that did not contain RT enzyme during RT synthesis or template DNA during PCR (not shown). RT–PCR was repeated three times, with similar results. Itgb3 expression was analyzed by densitometry and normalized to β-actin as described in Fig. 4. Average Itgb3/β-actin ratio in each group was: U 0.83 ± 0.17, Uc 0.50 ± 0.07, D 0.28 ± 0.06 and Dc 0.44 ± 0.04. There was a significant increase in Itgb3 mRNA in endometrium transfected with pcDNA4/MEIS1 compared with endometrium transfected with empty pcDNA4 DNA (P = 0.02). Levels of Itgb3 mRNA in endometrium transfected with pTER/Meis1 siRNA were significantly decreased when compared with endometrium transfected with control pTER (P = 0.006). (b) Immunohistochemistry of Itgb3 protein expression in the endometrium of mice transfected with either pcDNA4/MEIS1 or pTER/Meis1 siRNA plasmid. Shown are representative images of each group: uterine sections from mice transfected with pcDNA4/MEIS1 (U), with empty pcDNA4 vector (Uc), with pTER/Meis1 siRNA (D) or with non-targeting pTER vector (Dc). No staining was observed in a negative control without Itgb3 antibody as the primary antiserum (not shown). Scale bar = 100 µm. Statistical analysis of Itgb3 immunoreactivity was performed as described in the ‘Materials and Methods’ section. The data were analyzed using a two-tailed unpaired t-test. The immunoreactivity analysis was repeated three times, with similar results. The average Itgb3 immunoreactivity scores in each group were: U 6.4 ± 0.5, Uc 4.0 ± 0.8, D 2.4 ± 0.9 and Dc 4.8 ± 0.7. Uterine sections from pcDNA4/MEIS1-transfected mice showed significantly increased glandular and stromal Itgb3 expression compared with control mice treated with empty pcDNA4 vector (P = 0.0015). Uterine sections from pTER/Meis1 siRNA-transfected mice showed significantly reduced glandular and stromal Itgb3 expression compared with control mice treated with non-targeting pTER vector (P = 0.03)
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DEN082F7: Effect of Meis1 expression manipulation on murine uterine integrin β3 (Itgb3) expression.(a) RT–PCR analysis of Itgb3 expression in murine endometria transfected with pcDNA4/MEIS1 (U), empty pcDNA4 (Uc), pTER/Meis1 siRNA (D) or control pTER constructs (Dc). A representative agarose EtBr picture is shown. No PCR product was observed in control samples that did not contain RT enzyme during RT synthesis or template DNA during PCR (not shown). RT–PCR was repeated three times, with similar results. Itgb3 expression was analyzed by densitometry and normalized to β-actin as described in Fig. 4. Average Itgb3/β-actin ratio in each group was: U 0.83 ± 0.17, Uc 0.50 ± 0.07, D 0.28 ± 0.06 and Dc 0.44 ± 0.04. There was a significant increase in Itgb3 mRNA in endometrium transfected with pcDNA4/MEIS1 compared with endometrium transfected with empty pcDNA4 DNA (P = 0.02). Levels of Itgb3 mRNA in endometrium transfected with pTER/Meis1 siRNA were significantly decreased when compared with endometrium transfected with control pTER (P = 0.006). (b) Immunohistochemistry of Itgb3 protein expression in the endometrium of mice transfected with either pcDNA4/MEIS1 or pTER/Meis1 siRNA plasmid. Shown are representative images of each group: uterine sections from mice transfected with pcDNA4/MEIS1 (U), with empty pcDNA4 vector (Uc), with pTER/Meis1 siRNA (D) or with non-targeting pTER vector (Dc). No staining was observed in a negative control without Itgb3 antibody as the primary antiserum (not shown). Scale bar = 100 µm. Statistical analysis of Itgb3 immunoreactivity was performed as described in the ‘Materials and Methods’ section. The data were analyzed using a two-tailed unpaired t-test. The immunoreactivity analysis was repeated three times, with similar results. The average Itgb3 immunoreactivity scores in each group were: U 6.4 ± 0.5, Uc 4.0 ± 0.8, D 2.4 ± 0.9 and Dc 4.8 ± 0.7. Uterine sections from pcDNA4/MEIS1-transfected mice showed significantly increased glandular and stromal Itgb3 expression compared with control mice treated with empty pcDNA4 vector (P = 0.0015). Uterine sections from pTER/Meis1 siRNA-transfected mice showed significantly reduced glandular and stromal Itgb3 expression compared with control mice treated with non-targeting pTER vector (P = 0.03)

Mentions: Maternal Hoxa10 has earlier been shown to be involved in pinopode formation in the mouse uterus (Bagot et al., 2001), in a manner resembling the effect of Meis1 on pinopode formation observed in this study. To investigate a possible connection between Meis1 and Hoxa10 function in murine uterine development and embryonic implantation, we evaluated the effect of Meis1 expression manipulation on Itgb3 expression. To obtain a first indication of differential Itgb3 mRNA expression, peri-implantation uteri from mice treated with pcDNA4/MEIS1 or empty pcDNA4 (as a negative control), or with pTER/Meis1 or non-targeting pTER (as a negative control) were analyzed by RT–PCR. Fig. 7a shows a representative EtBr gel picture of the RT–PCR. Densitometric analysis was performed on all samples, and the average abundance of Itgb3 in each group normalized to β-actin was calculated. Levels of Itgb3 mRNA appeared significantly decreased in endometrium transfected with pTER/Meis1 siRNA compared with the control. In contrast, levels of Itgb3 mRNA increased in endometrium transfected with the pcDNA4/MEIS1 expression construct compared with the control (see legend to Fig. 7).


Myeloid ecotropic viral integration site 1 (MEIS) 1 involvement in embryonic implantation.

Xu B, Geerts D, Qian K, Zhang H, Zhu G - Hum. Reprod. (2008)

Effect of Meis1 expression manipulation on murine uterine integrin β3 (Itgb3) expression.(a) RT–PCR analysis of Itgb3 expression in murine endometria transfected with pcDNA4/MEIS1 (U), empty pcDNA4 (Uc), pTER/Meis1 siRNA (D) or control pTER constructs (Dc). A representative agarose EtBr picture is shown. No PCR product was observed in control samples that did not contain RT enzyme during RT synthesis or template DNA during PCR (not shown). RT–PCR was repeated three times, with similar results. Itgb3 expression was analyzed by densitometry and normalized to β-actin as described in Fig. 4. Average Itgb3/β-actin ratio in each group was: U 0.83 ± 0.17, Uc 0.50 ± 0.07, D 0.28 ± 0.06 and Dc 0.44 ± 0.04. There was a significant increase in Itgb3 mRNA in endometrium transfected with pcDNA4/MEIS1 compared with endometrium transfected with empty pcDNA4 DNA (P = 0.02). Levels of Itgb3 mRNA in endometrium transfected with pTER/Meis1 siRNA were significantly decreased when compared with endometrium transfected with control pTER (P = 0.006). (b) Immunohistochemistry of Itgb3 protein expression in the endometrium of mice transfected with either pcDNA4/MEIS1 or pTER/Meis1 siRNA plasmid. Shown are representative images of each group: uterine sections from mice transfected with pcDNA4/MEIS1 (U), with empty pcDNA4 vector (Uc), with pTER/Meis1 siRNA (D) or with non-targeting pTER vector (Dc). No staining was observed in a negative control without Itgb3 antibody as the primary antiserum (not shown). Scale bar = 100 µm. Statistical analysis of Itgb3 immunoreactivity was performed as described in the ‘Materials and Methods’ section. The data were analyzed using a two-tailed unpaired t-test. The immunoreactivity analysis was repeated three times, with similar results. The average Itgb3 immunoreactivity scores in each group were: U 6.4 ± 0.5, Uc 4.0 ± 0.8, D 2.4 ± 0.9 and Dc 4.8 ± 0.7. Uterine sections from pcDNA4/MEIS1-transfected mice showed significantly increased glandular and stromal Itgb3 expression compared with control mice treated with empty pcDNA4 vector (P = 0.0015). Uterine sections from pTER/Meis1 siRNA-transfected mice showed significantly reduced glandular and stromal Itgb3 expression compared with control mice treated with non-targeting pTER vector (P = 0.03)
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DEN082F7: Effect of Meis1 expression manipulation on murine uterine integrin β3 (Itgb3) expression.(a) RT–PCR analysis of Itgb3 expression in murine endometria transfected with pcDNA4/MEIS1 (U), empty pcDNA4 (Uc), pTER/Meis1 siRNA (D) or control pTER constructs (Dc). A representative agarose EtBr picture is shown. No PCR product was observed in control samples that did not contain RT enzyme during RT synthesis or template DNA during PCR (not shown). RT–PCR was repeated three times, with similar results. Itgb3 expression was analyzed by densitometry and normalized to β-actin as described in Fig. 4. Average Itgb3/β-actin ratio in each group was: U 0.83 ± 0.17, Uc 0.50 ± 0.07, D 0.28 ± 0.06 and Dc 0.44 ± 0.04. There was a significant increase in Itgb3 mRNA in endometrium transfected with pcDNA4/MEIS1 compared with endometrium transfected with empty pcDNA4 DNA (P = 0.02). Levels of Itgb3 mRNA in endometrium transfected with pTER/Meis1 siRNA were significantly decreased when compared with endometrium transfected with control pTER (P = 0.006). (b) Immunohistochemistry of Itgb3 protein expression in the endometrium of mice transfected with either pcDNA4/MEIS1 or pTER/Meis1 siRNA plasmid. Shown are representative images of each group: uterine sections from mice transfected with pcDNA4/MEIS1 (U), with empty pcDNA4 vector (Uc), with pTER/Meis1 siRNA (D) or with non-targeting pTER vector (Dc). No staining was observed in a negative control without Itgb3 antibody as the primary antiserum (not shown). Scale bar = 100 µm. Statistical analysis of Itgb3 immunoreactivity was performed as described in the ‘Materials and Methods’ section. The data were analyzed using a two-tailed unpaired t-test. The immunoreactivity analysis was repeated three times, with similar results. The average Itgb3 immunoreactivity scores in each group were: U 6.4 ± 0.5, Uc 4.0 ± 0.8, D 2.4 ± 0.9 and Dc 4.8 ± 0.7. Uterine sections from pcDNA4/MEIS1-transfected mice showed significantly increased glandular and stromal Itgb3 expression compared with control mice treated with empty pcDNA4 vector (P = 0.0015). Uterine sections from pTER/Meis1 siRNA-transfected mice showed significantly reduced glandular and stromal Itgb3 expression compared with control mice treated with non-targeting pTER vector (P = 0.03)
Mentions: Maternal Hoxa10 has earlier been shown to be involved in pinopode formation in the mouse uterus (Bagot et al., 2001), in a manner resembling the effect of Meis1 on pinopode formation observed in this study. To investigate a possible connection between Meis1 and Hoxa10 function in murine uterine development and embryonic implantation, we evaluated the effect of Meis1 expression manipulation on Itgb3 expression. To obtain a first indication of differential Itgb3 mRNA expression, peri-implantation uteri from mice treated with pcDNA4/MEIS1 or empty pcDNA4 (as a negative control), or with pTER/Meis1 or non-targeting pTER (as a negative control) were analyzed by RT–PCR. Fig. 7a shows a representative EtBr gel picture of the RT–PCR. Densitometric analysis was performed on all samples, and the average abundance of Itgb3 in each group normalized to β-actin was calculated. Levels of Itgb3 mRNA appeared significantly decreased in endometrium transfected with pTER/Meis1 siRNA compared with the control. In contrast, levels of Itgb3 mRNA increased in endometrium transfected with the pcDNA4/MEIS1 expression construct compared with the control (see legend to Fig. 7).

Bottom Line: Analysis of an Affymetrix data set in the public domain showed high expression of MEIS1 in human endometrium.MEIS1 expression was confirmed during the human menstrual cycle by RT-PCR and in situ hybridization and was increased during the secretory compared with proliferative phase of the cycle (P = 0.0001), the time of implantation.Increased or decreased Meis1 expression significantly increased or decreased the expression of integrin beta3, a transcriptional target of HOXA10 and an important factor in early embryo-endometrium interactions (P = 0.006).

View Article: PubMed Central - PubMed

Affiliation: Reproductive Medicine Center, Tongji Hospital, Tongji Medicine College, Huazhong, University of Science and Technology, 1095 JieFang Avenue, Wuhan 430030, People's Republic of China.

ABSTRACT

Background: The HOXA10 homeobox gene controls embryonic uterine development and adult endometrial receptivity. The three-amino-acid loop extension (TALE) family homeobox genes like myeloid ecotropic viral integration site 1 (MEIS) provide enhanced target gene activation and specificity in HOX-regulated cellular processes by acting as HOX cofactors.

Methods and results: Analysis of an Affymetrix data set in the public domain showed high expression of MEIS1 in human endometrium. MEIS1 expression was confirmed during the human menstrual cycle by RT-PCR and in situ hybridization and was increased during the secretory compared with proliferative phase of the cycle (P = 0.0001), the time of implantation. To assess the importance of maternal Meis1 expression in a mouse model, the uteri of Day 2 pregnant mice were injected with Meis1 over-expression or small interfering RNA (siRNA) constructs. Blocking Meis1 expression by siRNA before implantation significantly reduced average implantation rates (P = 0.00001). Increased or decreased Meis1 expression significantly increased or decreased the expression of integrin beta3, a transcriptional target of HOXA10 and an important factor in early embryo-endometrium interactions (P = 0.006). Manipulating Meis1 expression before implantation also dramatically affected the number of pinopodes, uterine endometrial epithelial projections that develop at the time of endometrial receptivity.

Conclusions: The results suggest that in mouse, meis1 contributes to regulating endometrial development during the menstrual cycle and establishing the conditions necessary for implantation.

Show MeSH
Related in: MedlinePlus