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长期饲喂生物柴油蓖麻工业残渣后山羊卵母细胞发育相关基因的比较表达谱

Comparative expression profiles of genes related to oocyte development in goats after long-term feeding with biodiesel castor industry residues.

作者信息

Silva L M, Lazzarotto C R, Tavares K C S, Oliveira C H A, Silva A M, Fernandes C C L, Aguiar L H, Martins L T, Neto S Gaudencio, Rios D B, Bertolini L R, Bertolini M, Silva C M G, Rondina D

机构信息

State University of Ceará, Fortaleza, Ceará, Brazil.

University of Fortaleza, Fortaleza, Ceará, Brazil.

出版信息

Anim Reprod Sci. 2014 Jul;148(1-2):32-41. doi: 10.1016/j.anireprosci.2014.05.009. Epub 2014 Jun 2.

Abstract

The aim of this study was to determine whether the consumption of detoxified castor meal (DCM) by goats over a long period of time affects mRNA levels in oocytes, and in mural granulosa and cumulus cells. A total of 41 adult does were supplemented (DCM group, n=21) or not (control group, n=20) with detoxified castor meal (DCM) for a period of 500 days. Then, 13 and 12 does were randomly selected for slaughter from the DCM and control treatments groups, respectively, for the determination of the number of visible ovarian follicles, retrieved cumulus-oocyte complexes (COCs), and viable and non-viable oocytes. The relative expression levels for distinct genes were determined by quantitative PCR in viable immature oocytes prior to in vitro maturation (IVM), in oocytes attaining or not the metaphase stage after IVM, as well as in granulosa cells obtained upon oocyte collection, and in cumulus cells obtained after IVM. The number of follicles ≥4 mm did not differ between treatments (overall mean 23.3 ± 2.0) and no significant differences were observed in the recovery of viable, non-viable, or total mean numbers of oocytes (control group: 44.7 ± 4.6, DCM group: 54.9 ± 5.9, respectively) between control and DCM fed goats. The maturation rate was significantly higher for control than DCM oocytes (58.0% vs. 45.3%; P<0.05). The mRNA levels in immature COC for controls were significantly higher for GLUT1 and lower for HSP70 (P<0.05) than for DCM. Following maturation, MII oocytes from both treatments had mRNA levels that were significantly higher for GDF9 and lower for BMP15 than for NC oocytes (P<0.05). In cumulus cells, the mRNA levels were significantly higher for LHR, FSHR, LeptinR, and IGF1, and lower for MnSOD in the control group compared with the DCM group (P<0.05). In conclusion, the inclusion of DCM in goat feed for long periods of time changed gene expression in immature oocytes and in cumulus cells. This was reflected by a decrease in the in vitro oocyte maturation rate.

摘要

本研究的目的是确定山羊长期食用脱毒蓖麻粕(DCM)是否会影响卵母细胞以及壁颗粒细胞和卵丘细胞中的mRNA水平。总共41只成年母羊在500天的时间里被补充(DCM组,n = 21)或未补充(对照组,n = 20)脱毒蓖麻粕(DCM)。然后,分别从DCM组和对照组中随机挑选13只和12只母羊进行屠宰,以确定可见卵巢卵泡的数量、回收的卵丘-卵母细胞复合体(COC)以及存活和非存活卵母细胞的数量。通过定量PCR测定未成熟卵母细胞在体外成熟(IVM)之前、IVM后达到或未达到中期阶段的卵母细胞、卵母细胞采集时获得的颗粒细胞以及IVM后获得的卵丘细胞中不同基因的相对表达水平。处理组之间≥4mm卵泡的数量没有差异(总体平均值23.3±2.0),并且在对照组和喂食DCM的山羊之间,在存活、非存活或卵母细胞总平均数量的回收方面未观察到显著差异(对照组:44.7±4.6,DCM组:54.9±5.9)。对照组卵母细胞的成熟率显著高于DCM组卵母细胞(58.0%对45.3%;P<0.05)。对照组未成熟COC中GLUT1的mRNA水平显著高于DCM组,而HSP70的mRNA水平显著低于DCM组(P<0.05)。成熟后,两种处理的MII期卵母细胞中GDF9的mRNA水平显著高于未成熟卵母细胞(NC),而BMP15的mRNA水平显著低于未成熟卵母细胞(P<0.05)。在卵丘细胞中,与DCM组相比对照组中LHR、FSHR、LeptinR和IGF1的mRNA水平显著更高,而MnSOD的mRNA水平更低(P<0.05)。总之,在山羊饲料中长期添加DCM会改变未成熟卵母细胞和卵丘细胞中的基因表达。这表现为体外卵母细胞成熟率的降低。

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