Massachusetts General Hospital, Harvard Stem Cell Institute, Harvard Medical School
This article is a part ofJoVE General. If you think this article would be useful for your research, please recommend JoVE to your institution's librarian.Recommend JoVE to Your Librarian
Current Access Through Your IP Address
Current Access Through Your Registered Email Address
Shea, K., Geijsen, N. Dissection of 6.5 dpc Mouse Embryos. J. Vis. Exp. (2), e160, doi:10.3791/160 (2007).
Analysis of gene expression patterns during early stages of mammalian embryonic development can provide important clues about gene function, cell-cell interaction and signaling mechanisms that guide embryonic patterning. However, dissection of the mouse embryo from the decidua shortly after implantation can be a challenging procedure, and detailed step-by-step documentation of this process is lacking.
Here we demonstrate how post-implantation (6.5 dpc) embryos are isolated by first dissecting the uterus of a pregnant mouse (detection of the vaginal plug was designated day 0.5 poist coitum) and subsequently dissecting the embryo from maternal decidua. The dissection of Reichert's membrane is described as well as the removal of the ectoplacental cone.
Dissection of reproductive organs from a female mouse
Primitive Streak-stage (6.5 dpc) Dissection
Postimplantation in the mouse occurs between 4.5 dpc and birth. This dissection method can be applied, essentially as described, to isolate earlier or later stage embryos at 5.5-8.5 dpc.
|Micro Dissecting Forceps serrated, straight||Tool||Roboz Surgical Instruments Co.||RS-5130||2 pairs|
|Micro Dissecting Tweezers Pattern #55 INOX||Tool||Roboz Surgical Instruments Co.||RS-5010||2 pairs|
|Dulbecco’s Phosphate Buffered Saline (1X)||Reagent||Invitrogen||14190|
|Fisherbrand Sterile Polysytrene Petri Dish||Tool||Fisher Scientific||08-757-12|
|4% Paraformaldehyde in PBS||Reagent|
|Straight Sharp-Pointed Dissecting Scissors||Tool||Fisher Scientific||08-940|
|Pregnant female mice||Animal|
1. Nagy, A. et. al. 2003. Manipulating the mouse embryo: A laboratory manual third edition. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.