Characterization of the canine 3beta-hydroxysteroid dehydrogenase and its expression in the corpus luteum during diestrus
| Autor: | Bernd Hoffmann, Steven D. Morley, Mariusz P. Kowalewski, Gerhard Schuler, Alexander F. Howie, JI Mason |
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| Rok vydání: | 2006 |
| Předmět: |
Gene isoform
Untranslated region DNA Complementary 17-Hydroxysteroid Dehydrogenases Endocrinology Diabetes and Metabolism Clinical Biochemistry Molecular Sequence Data Heterologous Biology Transfection Biochemistry Cell Line Substrate Specificity Endocrinology Dogs Rapid amplification of cDNA ends Corpus Luteum Complementary DNA medicine Animals Humans Amino Acid Sequence Molecular Biology In Situ Hybridization Base Sequence Cell Biology Diestrus Molecular biology Open reading frame medicine.anatomical_structure Real-time polymerase chain reaction Molecular Medicine Female Corpus luteum |
| Zdroj: | The Journal of steroid biochemistry and molecular biology. 101(4-5) |
| ISSN: | 0960-0760 |
| Popis: | 3beta-Hydroxysteroid dehydrogenase (3betaHSD) is a key enzyme in the synthesis of bioactive steroid hormones. Objectives of the present study were to clone canine 3betaHSD and to investigate its expression in dog corpora lutea (CL) covering the periods of their formation, early and late regression (days 5, 15, 25, 35, 45, 65 after ovulation). Complete complementary DNA sequence was amplified by RACE PCR. Subsequent cloning revealed that the canine ovarian 3betaHSD transcript was composed of a 5'-untranslated region (5'-UTR) of 126 nucleotides, an open reading frame (ORF) of 1122 nucleotides and a 3'-UTR of 441 nucleotides. The putative ORF encoded a 374 amino acid protein which remains highly conserved (79-85% identity) between species. The transient expression of the cloned canine 3betaHSD in a mammalian heterologous cell expression system (HEK293T cells) identified the 3betaHSD activity as the only activity of this canine enzyme (absence of any detectable 17-hydroxysteroid dehydrogenase activity). Qualitative RT-PCR revealed expression of 3betaHSD on all days investigated and the signals were strongest on days 5 and 15, with day 25 intensity tending to decrease. However, variability between individual animals was high. The significant decrease in the expression of 3betaHSD towards the end of diestrus as indicated by Real Time PCR (p |
| Databáze: | OpenAIRE |
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