Molecular characterization of bovine placental and ovarian 20?-hydroxysteroid dehydrogenase.

ABSTRACT: The enzyme 20?-hydroxysteroid dehydrogenase (20?-HSD) catalyzes the conversion of progesterone to its inactive form, 20?-hydroxyprogesterone. This enzyme plays a critical role in the regulation of luteal function in female mammals. In this study, we conducted the characterization and functional analyses of bovine 20?-HSD from placental and ovarian tissues. The nucleotide sequence of bovine 20?-HSD showed significant homology to that of goats (96%), humans (84%), rabbits (83%), and mice (81%). The mRNA levels increased gradually throughout the estrous cycle, the highest being in the corpus luteum (CL) 1 stage. Northern blot analysis revealed a 1.2? kb mRNA in the bovine placental and ovarian tissues. An antibody specific to bovine 20?-HSD was generated in a rabbit immunized with the purified, recombinant protein. Recombinant 20?-HSD protein produced in mammalian cells had a molecular weight of ?37? kDa. Bacterially expressed bovine 20?-HSD protein showed enzymatic activity. The expression pattern of the 20?-HSD protein in the pre-parturition placenta and the CL1 stage of the estrous cycle was similar to the level of 20?-HSD mRNA expression. Immunohistochemical analysis also revealed that bovine 20?-HSD protein was intensively localized in the large luteal cells during the late estrous cycle.

SUBMITTER: Naidansuren P 

PROVIDER: S-EPMC3207728 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature