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Increased expression of heat shock protein 105 in rat uterus of early pregnancy and its significance in embryo implantation.

Yuan JX, Xiao LJ, Lu CL, Zhang XS, Liu T, Chen M, Hu ZY, Gao F, Liu YX - Reprod. Biol. Endocrinol. (2009)

Bottom Line: Injection of antisense oligodeoxynucleotides to Hsp105 into pregnant rat uteri was carried out to look at effect of Hsp105 on embryo implantation.Furthermore, injection of antisense oligodeoxynucleotides to Hsp105 into the rat uterine horn on day 3 of pregnancy obviously suppressed the protein expression as expected and reduced number of the implanted embryos as compared with the control.Temporal and spatial changes in Hsp105 expression in pregnant rat uterus may play a physiological role in regulating embryo implantation.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, PR China. yuanjx@ioz.ac.cn

ABSTRACT

Background: Heat shock proteins (Hsps) are a set of highly conserved proteins, Hsp105, has been suggested to play a role in reproduction.

Methods: Spatio-temporal expression of Hsp105 in rat uterus during peri-implantation period was examined by immunohistochemistry and Western blot, pseudopregnant uterus was used as control. Injection of antisense oligodeoxynucleotides to Hsp105 into pregnant rat uteri was carried out to look at effect of Hsp105 on embryo implantation.

Results: Expression of Hsp105 was mainly in the luminal epithelium on day 1 of pregnancy, and reached a peak level on day 5, whereas in stroma cells, adjacent to the implanting embryo, the strongest expression of Hsp105 was observed on day 6. The immunostaining profile in the uterus was consistent with that obtained by Western blot in the early pregnancy. In contrast, no obvious peak level of Hsp105 was observed in the uterus of pseudopregnant rat on day 5 or day 6. Furthermore, injection of antisense oligodeoxynucleotides to Hsp105 into the rat uterine horn on day 3 of pregnancy obviously suppressed the protein expression as expected and reduced number of the implanted embryos as compared with the control.

Conclusion: Temporal and spatial changes in Hsp105 expression in pregnant rat uterus may play a physiological role in regulating embryo implantation.

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Fluorescence micrographs of rat uteri after intrauterine administration of FITC-ODNs. (A) The uterine horn treated with FITC- ODNs (either sense or antisense) displaying high level of fluorescence only in the luminal epithelium 2.5 hours after administration. (B) 48 hours later, a moderate fluorescence in the stromal compartment was observed after treatment with FITC- ODNs (either sense or antisense). (C) Uterine horn treated with unlabeled control A-ODNs displaying no fluorescence. le, luminal epithelium; s, stroma. Bar = 100 μm.
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Figure 5: Fluorescence micrographs of rat uteri after intrauterine administration of FITC-ODNs. (A) The uterine horn treated with FITC- ODNs (either sense or antisense) displaying high level of fluorescence only in the luminal epithelium 2.5 hours after administration. (B) 48 hours later, a moderate fluorescence in the stromal compartment was observed after treatment with FITC- ODNs (either sense or antisense). (C) Uterine horn treated with unlabeled control A-ODNs displaying no fluorescence. le, luminal epithelium; s, stroma. Bar = 100 μm.

Mentions: Using an A-ODNs as a blocker we examined effect of blockage of Hsp105 gene expression on rat implantation. To assess A-ODNs penetrating capacity, the Hsp105 FITC-ODNs was first injected into the uterine lumen, and then the uteri were taken for preparing sections at the indicated time points for FITC-ODNs examination by fluorescence microscopy. Strong green fluorescence representing cellular uptake of FITC-ODNs was observed in the luminal epithelium at 2.5 hours after injection (Fig. 5A). A detectable fluorescence in the underlying stroma was detected 48 hours later (Fig. 5B), indicating the penetration of Hsp105 ODNs into these cells in vivo. No fluorescence was observed in the contralateral horn treated with the unlabeled ODNs as the control (Fig. 5C).


Increased expression of heat shock protein 105 in rat uterus of early pregnancy and its significance in embryo implantation.

Yuan JX, Xiao LJ, Lu CL, Zhang XS, Liu T, Chen M, Hu ZY, Gao F, Liu YX - Reprod. Biol. Endocrinol. (2009)

Fluorescence micrographs of rat uteri after intrauterine administration of FITC-ODNs. (A) The uterine horn treated with FITC- ODNs (either sense or antisense) displaying high level of fluorescence only in the luminal epithelium 2.5 hours after administration. (B) 48 hours later, a moderate fluorescence in the stromal compartment was observed after treatment with FITC- ODNs (either sense or antisense). (C) Uterine horn treated with unlabeled control A-ODNs displaying no fluorescence. le, luminal epithelium; s, stroma. Bar = 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2667524&req=5

Figure 5: Fluorescence micrographs of rat uteri after intrauterine administration of FITC-ODNs. (A) The uterine horn treated with FITC- ODNs (either sense or antisense) displaying high level of fluorescence only in the luminal epithelium 2.5 hours after administration. (B) 48 hours later, a moderate fluorescence in the stromal compartment was observed after treatment with FITC- ODNs (either sense or antisense). (C) Uterine horn treated with unlabeled control A-ODNs displaying no fluorescence. le, luminal epithelium; s, stroma. Bar = 100 μm.
Mentions: Using an A-ODNs as a blocker we examined effect of blockage of Hsp105 gene expression on rat implantation. To assess A-ODNs penetrating capacity, the Hsp105 FITC-ODNs was first injected into the uterine lumen, and then the uteri were taken for preparing sections at the indicated time points for FITC-ODNs examination by fluorescence microscopy. Strong green fluorescence representing cellular uptake of FITC-ODNs was observed in the luminal epithelium at 2.5 hours after injection (Fig. 5A). A detectable fluorescence in the underlying stroma was detected 48 hours later (Fig. 5B), indicating the penetration of Hsp105 ODNs into these cells in vivo. No fluorescence was observed in the contralateral horn treated with the unlabeled ODNs as the control (Fig. 5C).

Bottom Line: Injection of antisense oligodeoxynucleotides to Hsp105 into pregnant rat uteri was carried out to look at effect of Hsp105 on embryo implantation.Furthermore, injection of antisense oligodeoxynucleotides to Hsp105 into the rat uterine horn on day 3 of pregnancy obviously suppressed the protein expression as expected and reduced number of the implanted embryos as compared with the control.Temporal and spatial changes in Hsp105 expression in pregnant rat uterus may play a physiological role in regulating embryo implantation.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, PR China. yuanjx@ioz.ac.cn

ABSTRACT

Background: Heat shock proteins (Hsps) are a set of highly conserved proteins, Hsp105, has been suggested to play a role in reproduction.

Methods: Spatio-temporal expression of Hsp105 in rat uterus during peri-implantation period was examined by immunohistochemistry and Western blot, pseudopregnant uterus was used as control. Injection of antisense oligodeoxynucleotides to Hsp105 into pregnant rat uteri was carried out to look at effect of Hsp105 on embryo implantation.

Results: Expression of Hsp105 was mainly in the luminal epithelium on day 1 of pregnancy, and reached a peak level on day 5, whereas in stroma cells, adjacent to the implanting embryo, the strongest expression of Hsp105 was observed on day 6. The immunostaining profile in the uterus was consistent with that obtained by Western blot in the early pregnancy. In contrast, no obvious peak level of Hsp105 was observed in the uterus of pseudopregnant rat on day 5 or day 6. Furthermore, injection of antisense oligodeoxynucleotides to Hsp105 into the rat uterine horn on day 3 of pregnancy obviously suppressed the protein expression as expected and reduced number of the implanted embryos as compared with the control.

Conclusion: Temporal and spatial changes in Hsp105 expression in pregnant rat uterus may play a physiological role in regulating embryo implantation.

Show MeSH
Related in: MedlinePlus