Characterization of membrane progestin receptor subtypes in zebrafish, Danio rerio / by Richard Ned Hanna.

Author/creator Hanna, Richard Ned author.
Other author Zhu, Yong, degree supervisor.
Other author East Carolina University. Department of Biology.
Format Theses and dissertations
Production2005.
Description54 leaves : illustrations ; 28 cm
Supplemental ContentAccess via ScholarShip
Subjects

Summary Oocyte maturation in fish species is triggered by specific progestins called maturation inducing steroids. In this study, two recently discovered G protein-coupled receptor (GPCR) like membrane progesterone receptor isoforms (mPRa and mPRP) were shown to be rapidly and specifically activated by the maturation inducing steroid in zebrafish, 4-pregnene-17a,20P-diol-3-one (17a,20P), further implying both receptors are the membrane progestin receptors responsible for progestin induced oocyte maturation. The zebrafish mPRa, and a previously uncharacterized mPRP isoform, were stably expressed in a progesterone response deficient mammalian breast cancer cell line, MDA MB-231. Expression and surface localization ofthe receptors was verified by flow cytometry, biotin surface labeling and Western blotting. MAPK activation of the transfected cell lines was measured by Western blotting and flow cytometry after only 5 minutes stimulation by various steroids. Specific progestins were shown to significantly stimulate both receptors similarly with varying amounts of MAPK activation [17a,20p >progesterone (P4) >4-pregnen-17,20p,21-triol-3-one (20P-S)]. Furthermore, luM of 17a,20p was also shown to significantly decrease cAMP levels in both mPRa and mPRp transfected cells in a dose responsive and time dependent manner. Interestingly, the maturation inducing steroid in zebrafish, 17a,20p, was shown to be the most significant activator of MAPK and inhibitor ofcAMP for both receptors. This study, along with previous work, implies that both mPRa and mPRP are involved in similar functions and show a strong link to maturation inducing steroid provoked fish oocyte maturation.
General notePresented to the faculty of the Department of Biology.
General noteAdvisor: Yong Zhu
Dissertation noteM.S. East Carolina University 2005
Bibliography noteIncludes bibliographical references (leaves 46-51).
Genre/formAcademic theses.
Genre/formAcademic theses.
Genre/formThèses et écrits académiques.

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