Gene and protein expression signature of endometrial glandular and stromal compartments during the window of implantation
A significant biological pathway, the cell cycle,was identified along with signature protein staining patterns after gene expression of the receptive endometrium was determined via laser microdissected glands and stroma.
Gloria E. Evans, Ph.D., José A. Martínez-Conejero, Ph.D., Gregory T.M. Phillipson, M.B., Ch.B., Carlos Simón, M.D., Ph.D., Les A. McNoe, M.Sc., Peter H. Sykes, M.B., Ch.B., José A. Horcajadas, Ph.D., Enid Y.N. Lam, Ph.D., Cristin G. Print, Ph.D., Iris L. Sin, Ph.D., John J. Evans, Ph.D.
Volume 97, Issue 6 , Pages 1365-1373.e2, June 2012
To map the changes in messenger RNA (mRNA) and protein abundance during the window of implantation in specifically endometrial stromal and glandular epithelial cells obtained using laser microdissection microscopy (LDM).
Endometrial samples were collected from two menstrual cycles at 2 and 7 days after first significant rise in blood LH, and separate cell populations were obtained using LDM. A new generation linear polymerase chain reaction (PCR) amplified the mRNA, which were hybridized to both Affymetrix U133 Plus2 and Agilent 4x44K microarrays followed by gene set analysis. Immunohistochemistry assessed protein expression between the two collection times.
In vitro fertilization clinic.
Nine Caucasian, fertile, cycling women.
Main Outcome Measure(s):
Cycle dating using blood markers; microarrays on laser microdissected glands and stroma; dual platform microarray confirmation; immunohistochemical analysis of cell cycle proteins.
The two microarray platforms showed concordance. During the window of implantation, a statistically significant network of 22 mRNA associated with the cell cycle was down-regulated. Immunohistochemistry identified altered localization in stroma.
Microarrays demonstrated glands and stroma have distinct mRNA signatures, each dependent on the day of the cycle. We characterized two compartments of the receptive endometrium with a transcriptomic signature identifying regulation of only the cell cycle. Immunohistochemical analysis of cell cycle proteins identified a signature staining pattern of nuclear relocalization of a group of cyclins of stromal cells, which may be clinically applicable.