Sugimura Satoshi, Yamanouchi Tadayuki, Palmerini Maria Grazia, Hashiyada Yutaka, Imai Kei, Gilchrist Robert B
Department of Biological Production, Institute of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan.
National Livestock Breeding Center, Fukushima 961-8511, Japan.
J Reprod Dev. 2018 Jun 22;64(3):233-241. doi: 10.1262/jrd.2018-009. Epub 2018 Mar 2.
The administration of follicle-stimulating hormone (FSH) prior to oocyte retrieval improves oocyte developmental competence. During bovine embryo production in vitro, however, oocytes are typically derived from FSH-unprimed animals. In the current study, we examined the effect of pre-in vitro maturation (IVM) with cAMP modulators, also known as the second messengers of FSH, on the developmental competence of oocytes derived from small antral follicles (2-4 mm) of FSH-unprimed animals. Pre-IVM with N6,2'-O-dibutyryladenosine 3',5'-cyclicmonophosphate (dbcAMP) and 3-isobutyl-1-methylxanthine (IBMX) for 2 h improved the blastocyst formation in oocytes stimulated by FSH or amphiregulin (AREG). Furthermore, pre-IVM enhanced the expression of the FSH- or AREG-stimulated extracellular matrix-related genes HAS2, TNFAIP6, and PTGS2, and epidermal growth factor (EGF)-like peptide-related genes AREG and EREG. Additionally, pre-IVM with dbcAMP and IBMX enhanced the expression of EGFR, and also increased and prolonged cumulus cell-oocyte gap junctional communication. The improved oocyte development observed using the pre-IVM protocol was ablated by an EGF receptor phosphorylation inhibitor. These results indicate that pre-IVM with cAMP modulators could contribute to the acquisition of developmental competence by bovine oocytes from small antral follicles through the modulation of EGF receptor signaling and oocyte-cumulus/cumulus-cumulus gap junctional communication.
在取卵前施用促卵泡激素（FSH）可提高卵母细胞的发育能力。然而，在牛体外胚胎生产过程中，卵母细胞通常来自未用FSH预处理的动物。在本研究中，我们研究了用环磷酸腺苷（cAMP）调节剂（也称为FSH的第二信使）进行体外成熟（IVM）预处理对来自未用FSH预处理动物的小窦卵泡（2-4毫米）卵母细胞发育能力的影响。用N6,2'-O-二丁酰腺苷3',5'-环一磷酸（dbcAMP）和3-异丁基-1-甲基黄嘌呤（IBMX）进行2小时的IVM预处理可提高FSH或双调蛋白（AREG）刺激的卵母细胞的囊胚形成率。此外，IVM预处理增强了FSH或AREG刺激的细胞外基质相关基因HAS2、TNFAIP6和PTGS2以及表皮生长因子（EGF）样肽相关基因AREG和EREG的表达。此外，用dbcAMP和IBMX进行IVM预处理增强了表皮生长因子受体（EGFR）的表达，还增加并延长了卵丘细胞-卵母细胞间隙连接通讯。使用IVM预处理方案观察到的卵母细胞发育改善被一种EGF受体磷酸化抑制剂消除。这些结果表明，用cAMP调节剂进行IVM预处理可能通过调节EGF受体信号传导以及卵母细胞-卵丘/卵丘-卵丘间隙连接通讯，有助于未用FSH预处理动物的小窦卵泡中的牛卵母细胞获得发育能力。
