Development of functional LH Receptors on pig cumulus-oocyte complexes cultured in vitro by a novel two-step culture system
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
19382213
DOI
10.1002/mrd.21039
Knihovny.cz E-resources
- MeSH
- Analysis of Variance MeSH
- Cell Culture Techniques methods MeSH
- Bucladesine pharmacology MeSH
- Gene Expression drug effects MeSH
- Follicle Stimulating Hormone pharmacology MeSH
- Glucuronosyltransferase biosynthesis genetics metabolism MeSH
- Cumulus Cells cytology drug effects metabolism MeSH
- Hyaluronic Acid metabolism MeSH
- Luteinizing Hormone pharmacology MeSH
- RNA, Messenger biosynthesis genetics MeSH
- Oocytes cytology drug effects growth & development MeSH
- Ovarian Follicle drug effects metabolism MeSH
- Swine MeSH
- Progesterone metabolism MeSH
- Cyclic AMP-Dependent Protein Kinases metabolism MeSH
- Receptors, LH biosynthesis genetics metabolism MeSH
- Animals MeSH
- Check Tag
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Bucladesine MeSH
- Follicle Stimulating Hormone MeSH
- Glucuronosyltransferase MeSH
- Hyaluronic Acid MeSH
- Luteinizing Hormone MeSH
- RNA, Messenger MeSH
- Progesterone MeSH
- Cyclic AMP-Dependent Protein Kinases MeSH
- Receptors, LH MeSH
We show in the present study that freshly isolated pig cumulus-oocyte complexes (COCs) display a limited response to LH, as assessed by the expression of hyaluronan synthase 2 (Has2) mRNA, activation of protein kinase A (PKA), production of hyaluronic acid (HA) and progesterone, cumulus cell expansion and resumption of meiosis. These data indicate that freshly isolated COCs do not possess a sufficient number of functional LH receptors (LHR). However, the expression of Lhr significantly increased during the culture of COCs in vitro in a medium supplemented with FSH. Assuming that the effect of FSH on LHR induction is mediated via cAMP signaling pathways, we developed a new culture system, in which the COCs were pre-cultured for 72 hr in a medium supplemented with dbcAMP. The pre-cultured COCs remained in the germinal vesicle stage, their cumulus investment underwent a dramatic increase in size and gap junctions between the cumulus cells were preserved. The stimulation of such COCs with either FSH or LH led to the resumption and completion of meiosis, activation of PKA, expression of Has2, synthesis of large amounts of HA and progesterone, and extensive expansion of cumulus cells. We conclude that the formation of functional LHR is stimulated in cumulus cells during the culture in vitro in a cAMP-dependent pathway. The dbcAMP-treated COCs thus represent a new model in which the resumption of meiosis and cumulus expansion can be induced exclusively by the action of recombinant LH.
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