| First Author | Kaye PL | Year | 1996 |
| Journal | J Reprod Fertil | Volume | 107 |
| Pages | 6-7 (Abstr.) | Mgi Jnum | J:33815 |
| Mgi Id | MGI:81308 | Citation | Kaye PL, et al. (1996) Early pregnancy factor/chaperonin 10 in preimplantation mouse embryos. J Reprod Fertil 107:6-7 (Abstr.) |
| abstractText | Full text of Abstract: 11. Early pregnancy factor/chaperonin 10 in preimplantation mouse embryos. PL Kaye1, CM Corcoranl, M Pantaleon1, AC Cavanagh2, H Morton2. Departments of 1Physiology & Pharmacology and 2Surgery, The University of Queensland, Brisbane, Qld 4072, Australia Early pregnancy factor (EPF) is required for successful establishment of pregnancy in vivo. Recent identification of EPF as a homologue of chaperonin 10 and its cloning and synthetic production have provided precise molecular tools with which to investigate its role in early pregnancy. This study reports the presence and location of EPF/cpn10 in preimplantation mouse embryos using antibodies raised against synthetic EPF-derived peptides. Embryos from superovulated Quackenbush mice were lysed and the proteins separated by polyacrylamide gel electrophoresis, transferred to membranes and probed with antiserum against an N-terminal peptide of EPF using the ECL system (Amersham). Pure EPF ran as a distinct band with Mr about 10 kDa. A similarly mobile band was present in lysates of fertilised and unfertilised eggs, 2-cell embryos, morulae and blastocysts. To identify the cellular location of this EPF/cpn10, fixed whole blastocysts were probed with an antibody directed against the C-terminal portion of EPF and stained with texas red linked species specific secondary antibody. These blastocysts revealed intense staining in nuclear and perinuclear locations of trophectoderm cells with little staining of inner cell mass cells. Since cpn10 is expressed in mitochondria which are present in preimplantation embryos, it is not surprising that antibody reactivity was observed in all stages; however the intra-cellular location in blastocysts does not correlate with a mitochondrial location. Furthermore, the cells expressing most EPF/cpn10 in blastocysts were predominantly in the trophectoderm, as would be expected from the observation that blastocysts secrete EPF. Critical in resolving this issue will be demonstration of secretion of EPF into the medium. |
