Systematic determination of differential gene expression in the primate corpus luteum during the luteal phase of the menstrual cycle.
The molecular and cellular processes required for development, function, and regression of the primate corpus luteum (CL) are poorly defined. We hypothesized that there are dynamic changes in gene expression occurring during the CL life span, which represent proteins and pathways critical to its regulation. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during the luteal phase of natural menstrual cycles. CL were collected between d 3-5 (early stage), d 7-8 (mid), d 10-12 (mid-late), d 14-16 (late), or d 18-19 (very-late) after the midcycle LH surge. From the early through very-late stages, 3234 transcripts were differentially expressed, with 879 occurring from the early through late stages that encompass the processes of luteinization, maintenance, and functional regression. To characterize gene changes most relevant to these processes, ontology analysis was performed using the list of 879 differentially expressed transcripts. Four main groups of related genes were identified with relevance to luteal physiology including: 1) immune function; 2) hormone and growth factor signaling; 3) steroidogenesis; and 4) prostaglandin biosynthesis, metabolism, and signaling. A subset of genes representing each of the four major categories was selected for validation of microarray results by quantitative real-time PCR. Results in mRNA levels were similar between the two methodologies for 17 of 18 genes. Additionally, protein levels for three genes were determined by Western blot analysis to parallel mRNA levels. This database will facilitate the identification of many novel or previously underappreciated pathways that regulate the structure and function of the primate CL.
Bogan
Randy L
RL
Division of Reproductive Sciences, Oregon National Primate Research Center, Oregon Health and Science University West Campus, Beaverton, Oregon 97006, USA.
Murphy
Melinda J
MJ
Stouffer
Richard L
RL
Hennebold
Jon D
JD
D000818:Animals; D015153:Blotting, Western; D003338:Corpus Luteum / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D008183:Luteal Phase; D008253:Macaca mulatta / Q000235:genetics*; D008597:Menstrual Cycle; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011981:Receptors, Prolactin / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D011981:Receptors, Prolactin