[1]王秋实,汪琴,韦伟,等.ACSF2启动子区c.-751 A>C突变影响扬州鹅产蛋性能作用机制研究[J].南京农业大学学报,2021,44(3):533-540.[doi:10.7685/jnau.202002028]
 WANG Qiushi,WANG Qin,WEI Wei,et al.The research on the mechanism of the effect of c.-751 A>C mutation in ACSF2 promoter on egg-laying performance of Yangzhou goose[J].Journal of Nanjing Agricultural University,2021,44(3):533-540.[doi:10.7685/jnau.202002028]
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ACSF2启动子区c.-751 A>C突变影响扬州鹅产蛋性能作用机制研究()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
44卷
期数:
2021年3期
页码:
533-540
栏目:
动物科学
出版日期:
2021-05-10

文章信息/Info

Title:
The research on the mechanism of the effect of c.-751 A>C mutation in ACSF2 promoter on egg-laying performance of Yangzhou goose
作者:
王秋实 汪琴 韦伟 张鑫宝 夏梦圆 张立凡 陈杰
南京农业大学动物科技学院, 江苏 南京 210095
Author(s):
WANG Qiushi WANG Qin WEI Wei ZHANG Xinbao XIA Mengyuan ZHANG Lifan CHEN Jie
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
关键词:
ACSF2SNP启动子扬州鹅产蛋性能
Keywords:
ACSF2SNPpromoterYangzhou gooselaying performance
分类号:
S835
DOI:
10.7685/jnau.202002028
摘要:
[目的] 本试验旨在研究酰基辅酶A合成酶家族成员2(ACSF2)基因启动子区突变c.-751 A>C与扬州鹅产蛋性能的相关性,并分析该突变位点对基因表达的调控作用。[方法] 利用等位基因特异PCR (AS-PCR)在343个扬州鹅个体中对c.-751 A>C位点进行基因型分析,并与产蛋性能关联分析;利用荧光定量PCR (qPCR)和荧光素酶表达载体检测AA型和CC型启动子活性差异;通过ACSF2过表达试验分析该基因对颗粒细胞能量代谢途径及鹅产蛋性能的影响。[结果] AA基因型个体产蛋性能显著优于CC基因型个体(P<0.05),与AC基因型个体的产蛋性能没有显著差异;qPCR结果表明AA基因型个体卵巢组织中ACSF2基因mRNA表达量显著低于CC基因型个体(P<0.01);荧光素酶表达载体的转录活性检测结果同样表明AA型启动子活性显著低于CC型。在扬州鹅卵泡颗粒细胞中过表达ACSF2基因,参与能量代谢的基因表达发生显著变化,ATP浓度显著升高(P<0.01),暗示ACSF2能够调控颗粒细胞内的能量代谢途径。[结论] ACSF2启动子区c.-751 A>C变异能够改变该基因的表达水平,并通过改变颗粒细胞能量代谢来影响扬州鹅的产蛋量。该突变位点可作为扬州鹅产蛋性能选育的分子标记。
Abstract:
[Objectives] This study aimed to verify the correlation between the c.-751 A>C in ACSF2(encoded by acyl-CoA synthetase family member 2) promoter and laying performance of Yangzhou goose,and analyze the effect of the SNP on ACSF2 expression. [Methods] We used allele specific PCR(AS-PCR) to analyze the genotype of SNP c.-751 A>C in a 343 Yangzhou geese population,and performed the correlation analysis between genotype and laying performance of Yangzhou goose. Quantitative real-time PCR(qPCR) and luciferase reporter gene vector were used to detect the difference between AA and CC promoter activity. ACSF2 over-expression was performed to analyze its role in the energy metabolism of granulosa cells and laying performance. [Results] Genotyping and association analysis results showed that the egg numbers of geese with AA genotype were significantly higher than those CC ones. Quantitative real-time PCR analysis indicated that AA individuals showed lower ACSF2 expression level in ovary tissues. Moreover,the overexpression experiment indicated that ACSF2 was involved in the energy metabolism of granulosa cells. [Conclusions] We identified a functional mutation in the promoter region of ACSF2 affecting gene expression in Yangzhou goose,and indicated the role of ACSF2 in laying performance by regulating energy metabolism of granulosa cells. Our results provide insight into the improvement of goose laying performance by ACSF2 genetic variation selection.

参考文献/References:

[1] Atlante A,de Bari L,Bobba A,et al. Cytochrome c,released from cerebellar granule cells undergoing apoptosis or excytotoxic death,can generate protonmotive force and drive ATP synthesis in isolated mitochondria[J]. J Neurochem,2003,86(3):591-604.
[2] Ferrari D,Stepczynska A,Los M,et al. Differential regulation and ATP requirement for caspase-8 and caspase-3 activation during CD95- and anticancer drug-induced apoptosis[J]. The Journal of Experimental Medicine,1998,188(5):979-984.
[3] Hu Y,Benedict M A,Ding L,et al. Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis[J]. The EMBO Journal,1999,18(13):3586-3595.
[4] Alonso-Pozos I,Rosales-Torres A M,ávalos-Rodríguez A,et al. Mechanism of granulosa cell death during follicular atresia depends on follicular size[J]. Theriogenology,2003,60(6):1071-1081.
[5] Eppig J J,Chesnel F,Hirao Y,et al. Oocyte control of granulosa cell development:how and why[J]. Human Reproduction(Oxford,England),1997,12(Suppl 11):127-132.
[6] Peluso J J,Liu X F,Romak J. Progesterone maintains basal intracellular adenosine triphosphate levels and viability of spontaneously immortalized granulosa cells by promoting an interaction between 14-3-3σ and ATP synthaseβ/precursor through a protein kinase G-dependent mechanism[J]. Endocrinology,2007,148(5):2037-2044.
[7] Su Y Q,Sugiura K,Eppig J J. Mouse oocyte control of granulosa cell development and function:paracrine regulation of cumulus cell metabolism[J]. Seminars in Reproductive Medicine,2009,27(1):32-42.
[8] Yu S G,Chu W W,Zhang L F,et al. Identification of laying-related SNP markers in geese using RAD sequencing[J]. PLoS One,2015,10(7):e0131572.
[9] Watkins P A,Maiguel D,Jia Z Z,et al. Evidence for 26 distinct acyl-coenzyme A synthetase genes in the human genome[J]. Journal of Lipid Research,2007,48(12):2736-2750.
[10] Fujino T,Kondo J,Ishikawa M,et al. Acetyl-CoA synthetase 2,a mitochondrial matrix enzyme involved in the oxidation of acetate[J]. Journal of Biological Chemistry,2001,276(14):11420-11426.
[11] Gilbert A B,Evans A J,Perry M M,et al. A method for separating the granulosa cells,the basal Lamina and the theca of the preovulatory ovarian follicle of the domestic fowl(Gallus domesticus)[J]. Journal of Reproduction and Fertility,1977,50(1):179-181.
[12] Hu S Q,Liu H H,Pan Z X,et al. Molecular cloning,expression profile and transcriptional modulation of two splice variants of very low density lipoprotein receptor during ovarian follicle development in geese(Anser cygnoide)[J]. Animal Reproduction Science,2014,149(3/4):281-296.
[13] Cosenza G,Pauciullo A,Gallo D,et al. Genotyping at the CSN1S1 locus by PCR-RFLP and AS-PCR in a Neapolitan goat population[J]. Small Ruminant Research,2008,74(1/2/3):84-90.
[14] 杨涛. 鹅GnRH基因5’端调控区和外显子1 SNPs检测及与产蛋量的关系[D]. 合肥:安徽农业大学,2007,16-29. Yang T. SNPs in geese GnRH gene 5’-flank region and exon 1 and association with egg production[D]. Hefei:Anhui Agricultural University,2007:16-29(in Chinese with English abstract).
[15] 牛晓童. 鹅FSHβ基因部分序列克隆及外显子3 SNPs与产蛋性能相关性研究[D]. 合肥:安徽农业大学,2008,16-24. Niu X T. Cloning of partial sequence of FSHβ gene and exon3 association of SNPs with egg production in goose[D]. Hefei:Anhui Agricultural University,2008:16-24(in Chinese with English abstract).
[16] 胡彦竞科. 四川白鹅GnRH、GnIH基因克隆、多态性及其与产蛋量的关联性研究[D]. 重庆:西南大学,2017,23-45. HU Y J K. Cloning,polymorphism of GnRH and GnIH genes and their association with egg production in Sichuan white goose[D]. Chongqing:Southwest University,2017:23-45(in Chinese with English abstract).
[17] Yu S,Xia M,Alsiddig M A,et al. Molecular cloning,alternative splicing and mRNA expression analysis of MAGI1 and its correlation with laying performance in geese[J]. British Poultry Science,2017,58(2):158-165.
[18] Xia M Y,Wei W,Jiang Z H,et al. A functional mutation in KIAA1462 promoter decreases glucocorticoid receptor affinity and affects egg-laying performance in Yangzhou geese[J]. International Journal of Molecular Sciences,2018,19(5):1531.
[19] Glasemacher J,Bock A K,Schmid R,et al. Purification and properties of acetyl-CoA synthetase(ADP-forming),an archaeal enzyme of acetate formation and ATP synthesis,from the hyperthermophile Pyrococcus furiosus[J]. European Journal of Biochemistry,1997,244(2):561-567.
[20] Tang H Y,Han M. Fatty acids regulate germline sex determination through ACS-4-dependent myristoylation[J]. Cell,2017,169(3):457-469.
[21] Zhang T,Xi Q S,Wang D,et al. Mitochondrial dysfunction and endoplasmic Reticulum stress involved in oocyte aging:an analysis using single-cell RNA-sequencing of mouse oocytes[J]. Journal of Ovarian Research,2019,12(1):1-9.
[22] Hashimoto S,Yamanaka M,Yamochi T,et al. Mitochondrial function in immature bovine oocytes is improved by an increase of cellular cyclic AMP[J]. Scientific Reports,2019,9:5167.
[23] Heydarnejad A,Ostadhosseini S,Varnosfaderani S R,et al. Supplementation of maturation medium with CoQ10 enhances developmental competence of ovine oocytes through improvement of mitochondrial function[J]. Molecular Reproduction and Development,2019,86(7):812-824.
[24] Xu H Y,Geng S S,Li T T,et al. Maturation of buffalo oocytes in vitro with acetyl-L-carnitine improves cryotolerance due to changes in mitochondrial function and the membrane lipid profile[J]. Reproduction,Fertility,and Development,2019,31(2):386-394.
[25] Zou H J,Chen B L,Ding D,et al. Melatonin promotes the development of immature oocytes from the COH cycle into healthy offspring by protecting mitochondrial function[J]. Journal of Pineal Research,2020,68(1):e12621.
[26] 陈孚江,周景文,史仲平,等. 乙酰辅酶A合成代谢对酿酒酵母生理功能的影响[J]. 微生物学报,2010,50(9):1172-1179. Chen F J,Zhou J W,Shi Z P,et al. Effect of acetyl-CoA synthase gene overexpression on physiological function of Saccharomyces cerevisiae[J]. Acta Microbiologica Sinica,2010,50(9):1172-1179(in Chinese with English abstract).
[27] Alonso-Pozos I,Rosales-Torres A M,ávalos-Rodríguez A,et al. Mechanism of granulosa cell death during follicular atresia depends on follicular size[J]. Theriogenology,2003,60(6):1071-1081.
[28] Tai C J,Chang S J,Chien L Y,et al. Adenosine triphosphate induces activation of Caspase-3 in apoptosis of human granulosa-luteal cells[J]. Endocrine Journal,2005,52(3):327-335.
[29] Park D W,Cho T,Kim M R,et al. ATP-induced apoptosis of human granulosa luteal cells cultured in vitro[J]. Fertility and Sterility,2003,80(4):993-1002.
[30] Yu S,Wei W,Xia M,et al. Molecular characterization,alternative splicing and expression analysis of ACSF2 and its correlation with egg-laying performance in geese[J]. Animal Genetics,2016,47(4):451-462.

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备注/Memo

备注/Memo:
收稿日期:2020-02-25。
基金项目:江苏省农业重大新品种创制项目(PZCZ201738)
作者简介:王秋实,硕士研究生。
通信作者:陈杰,博士,教授,主要从事动物分子遗传育种研究,Email:jiechen@njau.edu.cn。
更新日期/Last Update: 1900-01-01