Uteroferrin (Uf), a progesterone and estrogen regulated uterine secretory glycoprotein is implicated in both fetal development and placental function in the sow. However, its exact roles and mechanism of action in fetal and placental tissues are unknown. This proposal seeks to examine the regulatory mechanisms underlying the temporal and tissue specific expression of Uf as a first step towards the elucidation of its biological significance in development. Towards this end, the regulation of Uf synthesis and secretion will be studied at the level of its gene using current methodologies in gene technology. The complementary DNA (cDNA) for Uf will be isolated from a Lambdagt 11 expression library prepared from porcine uterine endometrial poly (A+) RNA, by screening with sheep polyclonal antibody raised against purified Uf. This clone will be characterized by restriction endonuclease mapping and nucleotide sequence analysis and will be used to evaluate the size and relative abundance of Uf mRNA(s) in uterine endometria of sows during the estrous cycle and pregnancy. These levels of mRNA will then be compared to the Uf protein content in endometrial cells and to the amount of Uf protein secreted by these cells. This will provide information on the temporal regulation of Uf protein and its mRNA during fetal and placental development. The chromosomal gene for Uf will be isolated using cloned cDNA as a probe. Complete elucidation of its nucleotide sequence and genomic organization will allow for comparisons with genes coding for proteins of similar functions and/or regulation and will give insights into the possible mechanism of Uf regulation by sex steroids. Lastly the Uf gene will be expressed in mouse C127 cells or human endometrial cells using bovine papilloma virus as vector in order to delineate the contribution of estrogen and progesterone in the regulation of Uf gene expression and to define the specific genomic regions responsible for this differential regulation. Uteroferrin represents an excellent model system for studying the role of other uterine secretory proteins in fetal and placental development. The proposed work will provide a foundation for further understanding the biological significance of Uf and may clarify the mechanisms underlying maternal-fetal interactions in mammalian systems that are essential for fetal development.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29HD021961-03
Application #
3469336
Study Section
Human Embryology and Development Subcommittee 2 (HED)
Project Start
1986-07-01
Project End
1989-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Ohio State University
Department
Type
Earth Sciences/Resources
DUNS #
098987217
City
Columbus
State
OH
Country
United States
Zip Code
43210
Heard, Melissa E; Velarde, Michael C; Giudice, Linda C et al. (2015) Krüppel-Like Factor 13 Deficiency in Uterine Endometrial Cells Contributes to Defective Steroid Hormone Receptor Signaling but Not Lesion Establishment in a Mouse Model of Endometriosis. Biol Reprod 92:140
Simmen, Rosalia C M; Heard, Melissa E; Simmen, Angela M et al. (2015) The Krüppel-like factors in female reproductive system pathologies. J Mol Endocrinol 54:R89-R101
Pabona, John Mark P; Zhang, Daying; Ginsburg, David S et al. (2015) Prolonged pregnancy in women is associated with attenuated myometrial expression of progesterone receptor co-regulator Krüppel-like Factor 9. J Clin Endocrinol Metab 100:166-74
Heard, Melissa E; Simmons, Christian D; Simmen, Frank A et al. (2014) Krüppel-like factor 9 deficiency in uterine endometrial cells promotes ectopic lesion establishment associated with activated notch and hedgehog signaling in a mouse model of endometriosis. Endocrinology 155:1532-46
Pabona, John Mark P; Simmen, Frank A; Nikiforov, Mikhail A et al. (2012) Krüppel-like factor 9 and progesterone receptor coregulation of decidualizing endometrial stromal cells: implications for the pathogenesis of endometriosis. J Clin Endocrinol Metab 97:E376-92
Heard, Melissa E; Pabona, John Mark P; Clayberger, Carol et al. (2012) The reproductive phenotype of mice null for transcription factor Krüppel-like factor 13 suggests compensatory function of family member Krüppel-like factor 9 in the peri-implantation uterus. Biol Reprod 87:115
Simmons, Christian D; Pabona, John Mark P; Heard, Melissa E et al. (2011) Krüppel-like factor 9 loss-of-expression in human endometrial carcinoma links altered expression of growth-regulatory genes with aberrant proliferative response to estrogen. Biol Reprod 85:378-85
Simmons, C D; Pabona, J M P; Zeng, Z et al. (2010) Response of adult mouse uterus to early disruption of estrogen receptor-alpha signaling is influenced by Krüppel-like factor 9. J Endocrinol 205:147-57
Simmen, R C M; Pabona, J M P; Velarde, M C et al. (2010) The emerging role of Krüppel-like factors in endocrine-responsive cancers of female reproductive tissues. J Endocrinol 204:223-31
Pabona, J M P; Velarde, M C; Zeng, Z et al. (2009) Nuclear receptor co-regulator Krüppel-like factor 9 and prohibitin 2 expression in estrogen-induced epithelial cell proliferation in the mouse uterus. J Endocrinol 200:63-73