Effects of Midkine on the Gene Expression in Bovine Cumulus Cells
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Accession number;04A0022316
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| Title;Effects of Midkine on the Gene Expression in Bovine Cumulus Cells |
| Author;
IKEDA SHUNTARO
(Kyoto Univ., Graduate School of Agriculture, JPN)
SOMEKAWA SHINJI
(Kyoto Univ., Graduate School of Agriculture, JPN)
IMAI HIROSHI
(Kyoto Univ., Graduate School of Agriculture, JPN)
YAMADA MASAYASU
(Kyoto Univ., Graduate School of Agriculture, JPN)
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Journal Title;Shokuniku ni kansuru Josei Kenkyu Chosa Seika Hokokusho
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Journal Code:X0296A
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ISSN:
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VOL.21;NO.;PAGE.24-30(2003)
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| Figure&Table&Reference;FIG.4, TBL.1, REF.18 |
| Pub. Country;Japan |
| Language;Japanese |
| Abstract;We previously reported that midkine(MK) added to in vitro maturation(IVM) medium of bovine cumulus-enclosed oocytes(CEOs) enhances the developmental competence of the oocytes to the blastocyst stage after in vitro fertilization via its direct action to the surrounding cumulus cells. The present study was conducted to identify mRNAs which are more abundant in MK treated cumulus cells than in untreated control cells. Cumulus cells were obtained by mechanical stripping from CEOs collected from antral follicles(2-5mm in diameter) in slaughterhouse-derived bovine ovaries. Groups of 8*104 cumulus cells were cultured in 50.MU.l drops of synthetic oviduct fluid containing 2%(v/v) BME amino acid solution, 1%(v/v) MEM non-essential amino acid sclution, 0.5mg/ml polyvinylalcohol. 1.MU.g/ml estradiol-17.BETA., and 100IU/ml human chorionic gonadotropin with or without(control) 200ng/ml recombinant bovine MK for 24h in 5% CO2 in air at 39.DEG.C.. PolyA+RNA were extracted from pooled cumulus cells(3*106) in both groups and subjected to PCR-based suppression subtractive hybridization followed by differential screening with four kinds of probes i.e. subtracted cDNA of MK treated cumulus cells, subtracted cDNA of control cumulus cells, unsubtrated cDNA of MK treated cumulus cells, and unsubtracted cDNA of control cumulus cells. Twenty-four putative positive clones were obtained from subtracted cDNA of MK treated cumulus cells. Among them, five mRNAs were confirmed to be more abundant in MK treated cumulus cells than in the control cells by virtual Northern blot analysis. We are currently studying the relations between these genes and the enhancing effect of MK upon the oocyte competency via surrounding cumulus cells. (author abst.) |
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