We have cloned the bovine homologue of Mater (maternal antigen that embryos require) cDNA, potentially the first germ cell-specific maternal-effect gene in this species. The 3297 base-pair longest open reading frame encodes a putative protein of 1098 amino acids with a domain organization similar to its human counterpart. By reverse transcription coupled to polymerase chain reaction, we have analyzed the spatiotemporal expression of MATER, along with other potential markers of germ cells or oocytes: ZAR1 (zygotic arrest 1), GDF9 (growth and differentiation factor 9), BMP15 (bone morphogenetic protein 15), and VASA. In agreement with a preferential oocyte origin, MATER, ZAR1, GDF9, and BMP15 transcripts were detected in the oocyte itself at a much higher level than in the gonads, while no significant expression was detected in our panel of somatic tissues (uterus, heart, spleen, intestine, liver, lung, mammary gland, muscle). In situ hybridization confirmed oocyte-restricted expression of MATER and ZAR1 within the ovary, as early as preantral follicle stages. VASA was highly represented in the testis and the ovary, and still present in the oocyte from antral follicles. Maternal MATER, ZAR1, GDF9, and BMP15 transcripts persisted during oocyte in vitro maturation and fertilization and in preimplantation embryo until the five- to eight-cell or morula stage, but transcription was not reactivated at the time of embryonic genome activation.

BackgroundAdiponectin is an adipokine, mainly produced by adipose tissue. It regulates several reproductive processes. The protein expression of the adiponectin system (adiponectin, its receptors, AdipoR1 and AdipoR2 and the APPL1 adaptor) in bovine ovary and its role on ovarian cells and embryo, remain however to be determined.MethodsHere, we identified the adiponectin system in bovine ovarian cells and embryo using RT-PCR, immunoblotting and immunohistochemistry. Furthermore, we investigated in vitro the effects of recombinant human adiponectin (10 micro g/mL) on proliferation of granulosa cells (GC) measured by [3H] thymidine incorporation, progesterone and estradiol secretions measured by radioimmunoassay in the culture medium of GC, nuclear oocyte maturation and early embryo development.ResultsWe show that the mRNAs and proteins for the adiponectin system are present in bovine ovary (small and large follicles and corpus luteum) and embryo. Adiponectin, AdipoR1 and AdipoR2 were more precisely localized in oocyte, GC and theca cells. Adiponectin increased IGF-1 10(-8) M-induced GC proliferation (P < 0.01) but not basal or insulin 10(-8) M-induced proliferation. Additionally, adiponectin decreased insulin 10(-8) M-induced, but not basal or IGF-1 10(-8) M-induced secretions of progesterone (P < 0.01) and estradiol (P < 0.05) by GC. This decrease in insulin-induced steroidogenesis was associated with a decrease in ERK1/2 MAPK phosphorylation in GC pre-treated with adiponectin. Finally, addition of adiponectin during in vitro maturation affected neither the percentage of oocyte in metaphase-II nor 48-h cleavage and blastocyst day 8 rates.ConclusionsIn bovine species, adiponectin decreased insulin-induced steroidogenesis and increased IGF-1-induced proliferation of cultured GC through a potential involvement of ERK1/2 MAPK pathway, whereas it did not modify oocyte maturation and embryo development in vitro.

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