Inhibition of proteasomal proteolysis affects expression of extracellular matrix components and steroidogenesis in porcine oocyte-cumulus complexes
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.
PubMed
22032857
DOI
10.1016/j.domaniend.2011.09.003
PII: S0739-7240(11)00141-X
Knihovny.cz E-resources
- MeSH
- Extracellular Matrix drug effects metabolism MeSH
- Cysteine Proteinase Inhibitors pharmacology MeSH
- Proteasome Inhibitors * MeSH
- Cumulus Cells drug effects metabolism MeSH
- Real-Time Polymerase Chain Reaction veterinary MeSH
- Leupeptins pharmacology MeSH
- RNA, Messenger biosynthesis genetics MeSH
- Cell Adhesion Molecules biosynthesis MeSH
- Oocytes drug effects metabolism MeSH
- Swine MeSH
- Progesterone biosynthesis MeSH
- Proteasome Endopeptidase Complex metabolism MeSH
- Animals MeSH
- Check Tag
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Names of Substances
- benzyloxycarbonylleucyl-leucyl-leucine aldehyde MeSH Browser
- Cysteine Proteinase Inhibitors MeSH
- Proteasome Inhibitors * MeSH
- Leupeptins MeSH
- RNA, Messenger MeSH
- Cell Adhesion Molecules MeSH
- Progesterone MeSH
- Proteasome Endopeptidase Complex MeSH
Porcine oocyte-cumulus complexes (OCCs) form an expanded cumulus extracellular matrix (ECM) in response to gonadotropins during meiotic maturation. Essential components of ECM are hyaluronan (HA), tumor necrosis factor α-induced protein 6 (TNFAIP6) and heavy chains (HC) of interalpha-trypsin inhibitor. To form expanded cumulus ECM, intermediate complexes (TNFAIP6-HC) must bind to HA to allow HC transfer onto HA. Protein turnover by the ubiquitin-proteasome pathway is poorly characterized in this process. It is known that the specific proteasomal inhibitor MG132 prevents cumulus expansion and formation of ECM. To determine whether inhibition of proteasomal proteolysis with MG132 affects cumulus cell steroidogenesis and expression of the cumulus expansion-related components (hyaluronan synthase type 2, HAS2, TNFAIP6) we cultured porcine OCCs and granulosa cells (GCs) in a medium supplemented with FSH/LH. Methods performed included real-time reverse transcription PCR, immunofluorescence and RIAs. The expression of TNFAIP6 and HAS2 transcripts increased significantly after the stimulation of OCCs and GCs with FSH/LH. In contrast, treatment with MG132 reduced the expression of TNFAIP6 and HAS2. Hyaluronan was detected with biotinylated HA-binding proteins within FSH/LH-stimulated expanded OCCs but not in those treated with MG132. Progesterone production, although increased almost three times after OCCs stimulation with FSH/LH, was significantly suppressed by MG132. The FSH/LH-stimulated a 40-fold increase in progesterone secretion by GCs was inhibited in the presence of MG132. In conclusion, MG132 affects progesterone secretion and expression of cumulus expansion-related components by cumulus and GCs, suggesting the requirement of ubiquitin-proteasome pathway-regulated protein turnover for formation of ECM during cumulus expansion in the preovulatory period in the pig.
References provided by Crossref.org
The Biological Role of Hyaluronan-Rich Oocyte-Cumulus Extracellular Matrix in Female Reproduction
