445. Focal adhesions disassemble during early pregnancy in rat uterine epithelial cells

Abstract

Successful blastocyst implantation require uterine epithelial cells (UECs) to undergo the 'plasma membrane transformation' followed by the removal of these cells around the implantation sites. The present study investigated the distribution and expression of two principal focal adhesion proteins talin and paxillin in rat UECs during early pregnancy and their role in the loss of these cells at the time of implantation. Results from immunofluorescence microscopy have demonstrated a major distributional change of talin and paxillin in UECs where an intense basal staining of these proteins on day 1 of pregnancy was lost at time of implantation. This was consistent with the significant decrease in paxillin seen through western blotting analysis. Interestingly the amount of talin was not significantly different between day 1 of pregnancy and at the time of implantation with a calpain 2 mediated proteolytic fragment of talin seen on both of these days of pregnancy. Calpain 2 activity was further investigated through western blotting analysis and increases in the active form of calpain 2 at the time of implantation. These observations suggest that talin and paxillin have disassembled from the site of focal adhesions where talin is undergoing cleavage by active calpain 2 at the time of implantation. This allows UECs to become less adherent to the underlying basal lamina facilitating their removal during blastocyst invasion. Hence, disassembly of focal adhesions at the time of implantation is a critical event for successful implantation and placentation.