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在牛卵母细胞体外成熟过程中使用妊娠相关血浆蛋白-A 会增加 IGF-1,并影响尼洛丽牛卵丘细胞和胚胎的转录谱。

Use of pregnancy-associated plasma protein-A during oocyte in vitro maturation increases IGF-1 and affects the transcriptional profile of cumulus cells and embryos from Nelore cows.

机构信息

Universidade do Oeste Paulista, Presidente Prudente, São Paulo, Brazil.

Departamento de Farmacologia, Universidade Estadual Paulista (UNESP), Botucatu, São Paulo, Brazil.

出版信息

Mol Reprod Dev. 2019 Nov;86(11):1694-1704. doi: 10.1002/mrd.23259. Epub 2019 Aug 29.

Abstract

Insulin-like growth factor 1 (IGF-1) activity is established by the regulation of IGF binding protein activity, which blocks IGF-1 functions, whereas pregnancy-associated plasma protein-A (PAPP-A) improves IGF-1 bioavailability and facilitates binding to IGF receptors. To further extend our understanding of the effect of exogenous PAPP-A on bovine embryo production, we added this protein during in vitro maturation of cumulus-oocyte complexes (COCs); moreover, we assessed its effects on IGF-1 quantity in the maturation medium, embryonic yield and postwarming survival, blastocyst quality, and transcript abundance. Bovine COCs were matured in a serum-free medium, either with PAPP-A supplementation (100 ng/ml) or without (control). The treatment group produced higher IGF-1 concentrations in the maturation medium; however, showed no difference on cleavage, blastocysts rates, and embryonic survival 3 and 24 hr postcryopreservation. Regarding gene expression, VNN1 was upregulated, whereas AGPAT9, FASN, EGFR, HAS2, and IMPDH1 were downregulated in PAPP-A treated. PAPP-A treated, CPT2, DNMT3A, and TFAM were upregulated, whereas ATF4 and IFITM3 were downregulated. We concluded that although the addition of PAPP-A did not affect embryo yield and blastocyst survival, higher IGF-1 levels may affect embryo competence through differential expression of genes involved in lipid metabolism, oocyte competence, and mitochondrial function.

摘要

胰岛素样生长因子 1(IGF-1)的活性由 IGF 结合蛋白活性的调节所建立,IGF 结合蛋白会阻断 IGF-1 的功能,而妊娠相关血浆蛋白-A(PAPP-A)则可提高 IGF-1 的生物利用度并促进其与 IGF 受体结合。为了进一步深入了解外源性 PAPP-A 对牛胚胎生产的影响,我们在卵丘-卵母细胞复合物(COC)的体外成熟过程中添加了这种蛋白;此外,我们评估了其对成熟培养基中 IGF-1 数量、胚胎产量和冻融后存活、囊胚质量和转录丰度的影响。牛 COC 在无血清培养基中成熟,分为添加 PAPP-A(100ng/ml)组和不添加(对照组)。处理组在成熟培养基中产生的 IGF-1 浓度较高;然而,在卵裂率、囊胚率和冻融后 3 小时和 24 小时的胚胎存活率方面没有差异。关于基因表达,VNN1 上调,而 AGPAT9、FASN、EGFR、HAS2 和 IMPDH1 在 PAPP-A 处理下下调。CPT2、DNMT3A 和 TFAM 在 PAPP-A 处理下上调,而 ATF4 和 IFITM3 下调。我们得出结论,尽管添加 PAPP-A 并未影响胚胎产量和囊胚存活率,但较高的 IGF-1 水平可能通过影响参与脂质代谢、卵母细胞能力和线粒体功能的基因的差异表达来影响胚胎能力。

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