The detection and confirmation of 11 beta-hydroxysteroid dehydrogenase type 1 transcripts in human luteinized granulosa cells using RT-PCR and plasmid pUC18.
The activity of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) in human granulosa cells has been shown to be associated with the outcome of treatment following in-vitro fertilization and embryo transfer. There are two known isoforms of 11 beta-HSD which differ significantly in their actions and co-factor requirements. The net activity of 11 beta-HSD which differ significantly in their actions and co-factor requirements. The net activity of 11 beta-HSD within the human ovary is unclear, but may be of particular importance within the ovarian follicle in regulating possible glucocorticoid influences on the oocyte. This study presents preliminary information regarding establishment of techniques to identify transcripts of the 11 beta-HSD isoforms within human granulosa cells and human cumulus cells using reverse transcription-polymerase chain reaction. In view of the high expression of the type 1 11 beta-HSD isoform and the possibility of other 11 beta-HSD isoforms in the ovary, plasmid technology was used to confirm the technique specifically identifying the known isoforms.
Smith
M P
MP
University of Bristol, Division of Obstetrics and Gynaecology, St. Michael's Hospital, UK.
Keay
S D
SD
Hall
L
L
Harlow
C R
CR
Jenkins
J M
JM
D042842:11-beta-Hydroxysteroid Dehydrogenases; D019976:Cloning, Organism; D003338:Corpus Luteum / Q000502:physiology*; D005260:Female; D006107:Granulosa Cells / Q000201:enzymology*; D006801:Humans; D006913:Hydroxysteroid Dehydrogenases / Q000096:biosynthesis*; D007527:Isoenzymes / Q000096:biosynthesis*; D008184:Luteal Cells / Q000201:enzymology; D009865:Oocytes / Q000502:physiology; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic
D007527:Isoenzymes; D011994:Recombinant Proteins; D006913:Hydroxysteroid Dehydrogenases; D042842:11-beta-Hydroxysteroid Dehydrogenases