title: The in-vitro effects of cAMP and cGMP modulators on inter-cellular dye transfer and gene expression levels in rat cumulus cell--oocyte complexes.
abstract: Supplementation of in-vitro maturation medium with reagents that inhibit meiotic resumption whilst supporting normal function of cumulus cell-oocyte complexes (COC) is challenging. This study compared the in-vitro effects of synthetic and physiologically-relevant reagents on meiotic resumption, gap junction activity and gene expression of rat COC. Higher doses of forskolin reduced gap junction activity. Whilst addition of phosphodiesterase inhibitors initially promoted gap junction activity, this decreased with time in-vitro. Moreover despite oocytes remaining in meiotic arrest, there was a concomitant decline in expression of genes critical for oocyte maturation, and evidence of a reduction in overall transcription rate. Similarly, supplementing media with C-type natriuretic peptide and/or oestradiol delayed meiotic resumption and only initially maintained gap junction activity. In contrast, several key genes were stimulated and overall transcription rates remained constant with time in-vitro. In summary, supplementation of media with physiologically-relevant reagents may better enable normal functions of the COC.
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, PO Box 600, Wellington 6140, New Zealand.
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Campen
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, PO Box 600, Wellington 6140, New Zealand.
authors.forename: Zaramasina L
authors.initials: ZL
authors.lastname: Clark
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, PO Box 600, Wellington 6140, New Zealand.
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Olds
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, PO Box 600, Wellington 6140, New Zealand.
authors.forename: Kenneth P
authors.initials: KP
authors.lastname: McNatty
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, PO Box 600, Wellington 6140, New Zealand. Electronic address: janet.pitman@vuw.ac.nz.
authors.forename: Janet L
authors.initials: JL
authors.lastname: Pitman
mesh_terms: D000818:Animals; D002229:Carbenoxolone / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D004396:Coloring Agents / Q000378:metabolism*; D018031:Connexin 43 / Q000378:metabolism; D054885:Cumulus Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000242:Cyclic AMP / Q000494:pharmacology*; D006152:Cyclic GMP / Q000494:pharmacology*; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3 / Q000378:metabolism; D005110:Extracellular Space / Q000378:metabolism*; D005260:Female; D017629:Gap Junctions / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D056726:Genetic Association Studies; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: D004396:Coloring Agents; D018031:Connexin 43; C580592:Gja1 protein, rat; D010726:Phosphodiesterase Inhibitors; D012333:RNA, Messenger; D005576:Colforsin; D000242:Cyclic AMP; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; C517290:Pde3a protein, rat; D006152:Cyclic GMP; D002229:Carbenoxolone
keywords: Cumulus cells; In-vitro; Meiosis; Oocyte; cAMP; cGMP