Inhibition of proteasomal proteolysis affects expression of extracellular matrix components and steroidogenesis in porcine oocyte-cumulus complexes
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem, 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-zdroje
- MeSH
- extracelulární matrix účinky léků metabolismus MeSH
- inhibitory cysteinových proteinas farmakologie MeSH
- inhibitory proteasomu * MeSH
- kumulární buňky účinky léků metabolismus MeSH
- kvantitativní polymerázová řetězová reakce veterinární MeSH
- leupeptiny farmakologie MeSH
- messenger RNA biosyntéza genetika MeSH
- molekuly buněčné adheze biosyntéza MeSH
- oocyty účinky léků metabolismus MeSH
- prasata MeSH
- progesteron biosyntéza MeSH
- proteasomový endopeptidasový komplex metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- benzyloxycarbonylleucyl-leucyl-leucine aldehyde MeSH Prohlížeč
- inhibitory cysteinových proteinas MeSH
- inhibitory proteasomu * MeSH
- leupeptiny MeSH
- messenger RNA MeSH
- molekuly buněčné adheze MeSH
- progesteron MeSH
- proteasomový endopeptidasový komplex 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.
Citace poskytuje Crossref.org
The Biological Role of Hyaluronan-Rich Oocyte-Cumulus Extracellular Matrix in Female Reproduction
