title: Assessment of Protein Expression by Proximity Ligation Assay in the Nonhuman Primate Endometrium, Placenta, and Fetal Adrenal in Response to Estrogen.
abstract: In the field of protein biology, immunology-based techniques have been evolving for detection and quantification of protein levels, protein-protein interaction, and protein modifications in cells and tissues. The proximity ligation assay (PLA), a method of detection that combines immunologic and PCR-based approaches, was developed to overcome some of the drawbacks that are inherent to other detection methods. The PLA allows for very sensitive and discretely quantifiable measures of unmodified, native protein levels, and protein-protein interaction/modification complexes in situ in both fixed tissues and cultured cells. We describe herein the PLA method and its applicability to quantify the effects of estrogen on expression of angioregulatory factors, e.g., angiopoietin-1 (Ang-1) in the endometrium, vascular endothelial growth factor (VEGF) in the placenta, and melanocortin 2 receptor (MC2R)/accessory protein (MRAP) in the fetal adrenal of the nonhuman primate.
authors.affiliation: Department of Biology, Buena Vista University, Storm Lake, IA, 50588, USA.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Bonagura
authors.affiliation: Departments of Obstetrics, Gynecology, Reproductive Sciences and Physiology, Center for Studies in Reproduction, University of Maryland School of Medicine, Bressler Research Laboratories 11-019, 655 West Baltimore Street, Baltimore, MD, 21201, USA.
authors.forename: Jeffery S
authors.initials: JS
authors.lastname: Babischkin
authors.affiliation: Department of Physiological Sciences, Eastern Virginia Medical School, Norfolk, VA, 23507, USA.
authors.forename: Gerald J
authors.initials: GJ
authors.lastname: Pepe
authors.affiliation: Departments of Obstetrics, Gynecology, Reproductive Sciences and Physiology, Center for Studies in Reproduction, University of Maryland School of Medicine, Bressler Research Laboratories 11-019, 655 West Baltimore Street, Baltimore, MD, 21201, USA. ealbrecht@fpi.umaryland.edu.
authors.forename: Eugene D
authors.initials: ED
authors.lastname: Albrecht
mesh_terms: D000311:Adrenal Glands / Q000187:drug effects* / Q000196:embryology / Q000276:immunology / Q000378:metabolism; D042683:Angiopoietin-1 / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D004717:Endometrium / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D005455:Fluorescent Antibody Technique; D008856:Microscopy, Fluorescence; D009841:Oligonucleotides / Q000378:metabolism; D010215:Papio; D010920:Placenta / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011506:Proteins / Q000276:immunology / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D044103:Receptor, Melanocortin, Type 2 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D057188:Workflow
chemical_list: D042683:Angiopoietin-1; D000906:Antibodies; D004967:Estrogens; D009841:Oligonucleotides; D011506:Proteins; D044103:Receptor, Melanocortin, Type 2; D042461:Vascular Endothelial Growth Factor A
keywords: Adrenal; Endometrium; Estrogen; Immunocytochemistry; Immunofluorescence; Placenta; Proximity ligation assay