The over-expression of ERA in osteoblast-like cells, in vitro, 2000
Mills, Ishara A.
2000-2009
Previous investigations have shown that estrogen plays a significant role in the development and maintenance of bone in mammals. The abrupt loss of estrogen at menopause and subsequent bone loss supports this hypothesis. By over-expressing estrogen receptor -a (ERa) in osteoblast-like cells, this project begins to study the effects of estrogen on bone development and maintenance through the ERa signaling pathway. The specific aims were to clone an osteoblast-specific promoter (i.e., rat osteocalcin promoter) upstream of the ERa cDNA and introduce this construct into osteoblast-like cell lines (i.e. MC3T3-E1 and ROS 17/2.8 cells). The functionality of the osteoblast-specific ERa construct was investigated by utilizing an estrogen-responsive chloramphenical acetyltransferase (CAT) reporter construct, which contains an estrogen responsive element (ERE) upstream of the CAT gene. The basic regulation of the osteoblast -specific promoter in the osteoblast -like cells was determined by performing co-transfection assays using a CAT reporter system. The osteoblast-specific ERa construct was introduced into a C57Blk/J6 mouse by using transgenic technology. The creation of an osteoblast-specific ERa expressing mouse can then be used to measure over-expression of the ERa in a wild type genetic background with future breeding into an estrogen receptor-alpha knock out (a ERKO) background, enabling us to understand estrogens effect on bone development and osteoporosis through the ERa pathway, in vitro.
text
application/pdf
2000-05-01
thesis
Master of Science (MS)
Clark Atlanta University
Department of Biological Sciences
Kimbro, Kevin Sean
Georgia--Atlanta
http://hdl.handle.net/20.500.12322/cau.td:2000_mills_ishara_a
