Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 4 de 4
Filter
Add more filters










Database
Language
Publication year range
1.
Fertil Steril ; 102(1): 307-317.e7, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24837612

ABSTRACT

OBJECTIVE: To characterize the transcriptome of luminal epithelia (LE) of fertile secretory endometria and compare the results with those from glandular epithelia (GE). DESIGN: Endometrial samples were collected at 2 and 7 days after initial blood LH surge in separate menstrual cycles. LE were obtained with the use of laser microdissection. mRNA was amplified with the use of linear polymerase chain reaction and hybridized to Agilent 4×44 microarrays. Gene analysis was used to identify differentially expressed mRNAs. Immunohistochemistry was used to assess nine proteins. SETTING: One IVF clinic. PATIENT(S): Seven Caucasian fertile cycling women. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Cycle dating with the use of blood endocrinologic markers, microarrays of laser-microdissected LE, immunohistochemical analysis. RESULT(S): One hundred sixty-one (of 401) differentially expressed mRNAs in LE were identified from the metabolism pathway. Increased selective protein expression in LE at 7 days after initial LH surge was observed. LE mRNA expression was the converse of that in GE. The two cell types each had a different significant biologic pathway identified. CONCLUSION(S): Our results introduce a new concept that LE differentially expressed mRNAs are in the converse direction to that of GE, indicating different biologic processes despite the GE being continuous with the luminal monolayer. This probable distinction of biologic roles has not been noted previously. Further investigations must take cognizance of this observation.


Subject(s)
Endometrium/metabolism , Epithelial Cells/metabolism , Fertility , Menstrual Cycle , Female , Fertility/genetics , Gene Expression Profiling/methods , Genetic Markers , Humans , Immunohistochemistry , Laser Capture Microdissection , Menstrual Cycle/ethnology , Menstrual Cycle/genetics , Oligonucleotide Array Sequence Analysis , RNA, Messenger/metabolism , Time Factors , White People
2.
Fertil Steril ; 97(6): 1365-73.e1-2, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22480820

ABSTRACT

OBJECTIVE: 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). DESIGN: 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. SETTING: In vitro fertilization clinic. PATIENT(S): Nine Caucasian, fertile, cycling women. INTERVENTION(S): None. 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. RESULT(S): 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. CONCLUSION(S): 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.


Subject(s)
Embryo Implantation/genetics , Endometrium/physiology , Epithelial Cells/physiology , Fertilization in Vitro , Oligonucleotide Array Sequence Analysis , Stromal Cells/physiology , Adult , Endometrium/cytology , Female , Fertility/genetics , Gene Expression/physiology , Humans , Luteal Phase/genetics , Luteinizing Hormone/blood , Young Adult
3.
Acta Obstet Gynecol Scand ; 82(11): 997-1003, 2003 Nov.
Article in English | MEDLINE | ID: mdl-14616272

ABSTRACT

BACKGROUND: The factors that determine oocyte competency are poorly understood. It is believed that angiogenic factors are crucial. Modulation of these factors is therefore a central consideration. The adjacent association of cumulus cells to oocytes gives these cells a particular importance in relation to oocyte behavior. We report the effects of gonadotropins on the secretion of VEGF and leptin from cumulus cells; and the concentrations of the proteins in follicular fluid and their relationship to fertilization of oocytes in vitro. METHODS: The subjects were women undergoing intracytoplasmic sperm injection (ICSI). Oocytes and follicular fluid were collected. Leptin and vascular endothelial growth factor (VEGF) concentrations in incubation supernatants and follicular fluid and leptin in cell lysates were measured. Fertilization of corresponding oocytes were noted. In the present study, cells from individual follicles were incubated, as well as pooled cells from a woman. RESULTS: For the first time we demonstrated that VEGF release by human cumulus cells was modulated by gonadotropins in a dose-related, time-dependent manner, but no leptin was detected in either the supernatants after cumulus cell incubations or in cell lysates. Mean leptin levels were similar whether from follicles associated with eggs that were fertilized (14.4 +/- 1.1 ng/ml, mean +/- SEM) or not (12.4 +/- 1.1 ng/ml). Mean VEGF levels were also similar (11.1 +/- 1.2 ng/ml; 12.8 +/- 1.3 ng/ml). A greater proportion of VEGF in follicles was derived from follicular activities, compared with transfer from other physiological compartments, than leptin. CONCLUSIONS: Whereas it is possible that VEGF is part of the gonadotropin-mediated network that regulates development of the oocyte, leptin may not be produced in significant quantities by cumulus cells or be related to oocyte competency.


Subject(s)
Chorionic Gonadotropin/pharmacology , Follicle Stimulating Hormone/pharmacology , Leptin/metabolism , Oocytes/growth & development , Vascular Endothelial Growth Factor A/metabolism , Adult , Analysis of Variance , Cell Culture Techniques , Female , Fertilization in Vitro , Humans , Oocytes/cytology
4.
Int J Androl ; 26(1): 46-51, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12534937

ABSTRACT

This study investigated the relationship between variation in the polymorphic CAG trinucleotide repeat (TNR) region of the human androgen receptor (AR) gene and semen quality in a Caucasian sample population. These men were patients attending the New Zealand Centre for Reproductive Medicine in Christchurch. The AR TNR region was amplified by polymerase chain reaction and then DNA sequenced to determine exact numbers of CAG repeats for each sample. In addition, the samples were screened for microdeletions within the AZFc region of the Y-chromosome. A total of 105 men with poor semen quality were compared with a group of 93 men with normal semen quality. Men with poor semen quality had similar CAG repeat number to men with normal semen quality (21.46 +/- 0.30 vs. 20.99 +/- 0.28, p = 0.126). Y-chromosome microdeletions were only detected in men with suboptimal semen parameters (7.4%). However, the presence of a deletion was not related to CAG repeat number. The CAG repeat number in the men with normal semen quality in the present study is similar to the Australian and German samples, but lower than those reported for the Swedes, Dutch and Danes. These results are contrary to the hypothesis that higher CAG repeats are associated with infertility in men, but strongly suggest that different populations may show different numbers of CAG repeats in addition to racial variation reported in previous studies.


Subject(s)
Infertility, Male/genetics , Receptors, Androgen/genetics , Semen , Trinucleotide Repeat Expansion , Gene Deletion , Humans , Male , New Zealand , White People
SELECTION OF CITATIONS
SEARCH DETAIL
...