Author
ROMEREIM, SARAH - UNIVERSITY OF NEBRASKA | |
HOU, XIAOYING - NEBRASKA MEDICAL CENTER | |
TALBOTT, HEATHER - NEBRASKA MEDICAL CENTER | |
Cushman, Robert - Bob | |
WOOD, JENNIFER - UNIVERSITY OF NEBRASKA | |
DAVIS, JOHN - NEBRASKA MEDICAL CENTER | |
CUPP, ANDREA - UNIVERSITY OF NEBRASKA |
Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only Publication Acceptance Date: 3/6/2016 Publication Date: 7/16/2016 Citation: Romereim, S.M., Hou, X., Talbott, H., Cushman, R.A., Wood, J.R., Davis, J.S., Cupp, A.S. 2016. A comparison of ovarian follicular and luteal cell gene expression profiles provides insight into cellular identities and functions [abstract]. Society for the Study of Reproduction Annual Meeting. Abstract #258 (Scientific Program p. 181). Available: http://www.ssr.org/sites/ssr.org/files/uploads/attachments/node/320/2016_ssr_abstracts.pdf Interpretive Summary: Technical Abstract: After ovulation, somatic cells of the ovarian follicle (theca and granulosa cells) become the small and large luteal cells of the corpus luteum. Aside from known cell type-specific receptors and steroidogenic enzymes, little is known about the differences in the gene expression profiles of these four cell types. We hypothesized that a full analysis of the gene expression patterns of these cells would yield new cell-specific genetic markers, functional insight into the behavior of each cell type, and potentially lineage markers to determine which luteal cell type arose from the theca or granulosa cells. To address this, a comparison of the RNA present in each cell type [theca cells (TC, n=3), granulosa cells (GC, n=4), large luteal cells (LLC, n=3), and small luteal cells (SLC, n=3)] as measured by Affymetrix microarray analysis was performed and the biological relevance interpreted via Gene Ontology Annotations and Ingenuity Pathway Analysis of the differentially expressed RNAs. Microarrays were performed using cells isolated from large antral follicles of estrous-synchronized cows or corpora lutea from cows from a local abattoir. Each cell type had groups of genes with enriched expression compared to all three other cell types. The TC population had 166 differentially expressed RNAs (e.g. smooth muscle specific actin gamma2, hydroxyprostaglandin dehydrogenase-15); GC had 567 (e.g. FSH receptor, inhibin beta); LLC had 312 (e.g. prostaglandin F receptor, prolactin receptor, lipoprotein lipase); and SLC had 60 (e.g. LH receptor, CD274). Select genes have been validated with qPCR, with strong correlations between microarray and qPCR results. Analysis of the gene ontology and predicted roles of these genes revealed expected cell type-specific functions (such as increased cell growth/hypertrophy and vascularization in luteal cells) and less expected functions (such as immune response/inflammation and metabolism in large luteal cells). Principle Component Analysis of the gene expression profiles of these cell types also reveals that, based on principle component 1 (which accounts for 79.15% of the variance), theca cells and small luteal cells are closely related while granulosa cells and large luteal cells are also highly related. Thus, gene expression analysis revealed transcriptional markers for individual cell types, provided information on the regulation of cell behaviors relevant to ovarian function, and offers insight into the lineage and differentiation process that creates luteal cells from follicular cells. This research was supported by USDA/NIFA grants 2011-67015-20076 and 2013-67015-20965. |