Embryonic adhesion is not affected by endometrial leptin receptor gene silencing.
OBJECTIVE
In rodents, evidence suggests that the leptin system is mandatory for embryonic implantation. We aimed to investigate the functional relevance of the endometrial leptin receptor (OB-R) in the adhesion phase of human implantation.


METHODS
We used an in vitro model for embryonic adhesion, composed of a human endometrial cell line (HEC1-A) and B6C3F1 mouse embryos. The OB-R gene was silenced in a stable manner by RNA interference, and embryonic adhesion rates were analyzed.


METHODS
Research laboratory at a university-affiliated center.


METHODS
RNA interference.


METHODS
Embryonic adhesion in cells treated with OB-R RNAi.


RESULTS
The OB-R shRNA-transfected cells exhibited up to 80% lower OB-R mRNA levels than those of cells nontransfected or transfected with scrambled shRNA. The OB-R protein was also highly diminished in the stable OB-R shRNA-transfected HEC1A cells, whereas OAS1 expression was similar in both nontransfected and transfected cells. Embryonic adhesion rate was similar in nontransfected (94%) and HEC1-A transfected cells with a scrambled sequence (94%) or with OB-R silencing sequence (92%).


CONCLUSIONS
Knocking down the OB-R gene in a human endometrial cell line has no effect on the embryonic adhesion rate. Nevertheless, the functional relevance of this system can not be excluded in other phases of embryonic implantation, such as the invasion phase. Moreover, we describe a new approach to the functional analysis of candidate molecules implicated in blastocyst adhesion.
Cervero
Ana
A
Fundación IVI, Instituto Universitario IVI, Universidad de Valencia, Valencia, Spain.
Domínguez
Francisco
F
Horcajadas
Jose Antonio
JA
Quiñonero
Alicia
A
Pellicer
Antonio
A
Simón
Carlos
C
D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D010064:Embryo Implantation / Q000235:genetics; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D004717:Endometrium / Q000166:cytology / Q000196:embryology* / Q000378:metabolism*; D005260:Female; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D051379:Mice; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics*; D054411:Receptors, Leptin
C515448:LEPR protein, human; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin; C515447:leptin receptor, mouse