Studies for mechanisms of bovine marbling and developments of genetic markers

牛大理石花纹机制研究和遗传标记的开发

• 批准号: 21580351
• 负责人: MIZOGUCHI Yasushi
• 金额: $ 3.16万
• 依托单位: Meiji University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21580351/
• 关键词: 遺伝子; 応用動物; ゲノム; 畜産学; 育種学

To investigate genes involved in ruminants intramuscular adipogenesis, sixteen target genes(8 up and 8 down regulated genes) with dramatically changed expression(P<10^<-12>) were selected between differentiation and proliferation phases libraries in our previous SAGE studies of a clonal bovine intramuscular preadipocyte(BIP) cell line(Mizoguchi, 2010), with reference to 3T3-L1 cell lines as monogastric animals. We harvested the BIP and 3T3-L1 cells 0, 3, 6, 9 and 12 days after adipogenic stimulation containing all-trans retinoic acid(RA), which inhibits preadipocyte differentiation in non-ruminants and measured the gene expression levels by quantitative real-time PCR and the accumulation levels of triglyceride(TG). Fourteen genes expression pattern of the 16 genes during adipogenesis were confirmed the SAGE studies. The expression pattern of eleven genes clearly differed between 3T3-L1 and BIP cells. Simultaneously, we investigated the profiles of target genes by RA. TG accumulation was significantly induced(P<0.05) in BIP cells whereas completely inhibited(P<0.05) in 3T3-L1 cells by RA at day 6 after adipogenic stimulation. RA induced the expression levels of six genes and suppressed these of eight genes at day 3 after adipogenesis in BIPs(P<0.05 ; respectively). The effect of ADFP down-regulation on the accumulation of TG was examined by transfecting the BIP cells with ADFP siRNA. The expression level of ADFP in the ADFP-siRNA transfected cells was 21.4% of that in the scr-siRNA transfected cells in BIP 3 days after induction(P<0.05). Also, 6 days after induction, the accumulation of TG in the ADFP-siRNA transfected cells was 20% less than in the control cells(P<0.001). Our findings of gene expression profiles by RA will cause a favorable and detailed understanding in the molecular mechanisms of bovine intramuscular adipogenesis.

To investigate genes involved in ruminants intramuscular adipogenesis, sixteen target genes(8 up and 8 down regulated genes) with dramatically changed expression(P<10^<-12>) were selected between differentiation and proliferation phases libraries in our previous SAGE studies of a clonal bovine intramuscular preadipocyte(BIP) cell line(Mizoguchi, 2010), with reference to 3T3-L1 cell lines作为单胃动物。我们收集了含有全反式视黄酸（RA）的成脂刺激后0、3、6、9和12天的BIP和3T3-L1细胞，这些刺激抑制了非液体中的前脂肪细胞分化，并通过定量实时PCR和Triglyceride（TG）的积累水平（TG）测量了基因表达水平。在脂肪生成过程中，14个基因的14个基因表达模式证实了SAGE研究。 3T3-L1和BIP细胞之间的11个基因的表达模式显然有所不同。同时，我们通过RA研究了靶基因的曲线。在脂肪生成刺激后的第6天，RA在BIP细胞中显着诱导TG积累（P <0.05），而RA在3T3-L1细胞中完全抑制（P <0.05）。 RA诱导了六个基因的表达水平，并在BIPS脂肪形成后第3天抑制了这些基因的表达水平（分别为p <0.05；分别为p <0.05）。通过用ADFP siRNA转染BIP细胞，检查了ADFP下调对TG积累的影响。在诱导后3天，在BIP中，ADFP-SIRNA转染细胞中ADFP的表达水平为21.4％（P <0.05）。同样，在诱导后6天，TG在ADFP-SIRNA转染的细胞中的积累比对照细胞低20％（P <0.001）。我们通过RA对基因表达谱的发现将在牛肌内脂肪形成的分子机制中产生有利而详细的理解。

项目成果

西表島に生息するリュウキュウイノシシの遺伝的多様性解析手法の開発

西表岛琉球公猪遗传多样性分析方法的开发

• 发表时间: 2011
• 期刊: 明治大学農学部研究報告
• 作者: 坪井一真;村上慶太朗;石垣長健;渡辺信;溝口康
• 通讯作者: 溝口康

但馬牛における経済形質に関わるゲノム解析～鶴山土井におけるQTLの効果検証

但马牛经济性状相关基因组分析——鹤山土居QTL效应验证

• 发表时间: 2010
• 期刊: 兵庫県農林水産技術総合センター研究報告
• 作者: 秋山敬孝;龍田健;溝口康;西村翔太;杉本喜憲
• 通讯作者: 杉本喜憲

雌豚繁殖形質とPROP1遺伝子多型を用いた相関解析

母猪繁殖性状与PROP1基因多态性的相关分析

• 发表时间: 2012
• 作者: T. Yamada;S. Sasaki;S. Sukegawa;T. Miyake;T. Fujita;H. Kose;M. Morita;Y. Takahagi;H. Murakami;F. Morimatsu;Y. Sasaki;溝口康
• 通讯作者: 溝口康

DNAマーカーを指標とした牛の育種手法の開発に関する研究.岡山県基幹種雄牛「花茂勝2」におけるQTL領域の推定(第1報)

以DNA标记为指标的牛育种方法的开发研究，冈山县旗舰牛“花胜2号”的QTL区域估计（第1次报告）

• 发表时间: 2009
• 期刊: 岡山県総合畜産センター研究報告
• 作者: 中藤由紀;黒岩恵;馬場誠;溝口康;杉本喜憲
• 通讯作者: 杉本喜憲

ウシ脂肪交雑QTL(Marbling-3)の責任SNPの検出

检测负责牛大理石纹 QTL 的 SNP (Marbling-3)

• 发表时间: 2009
• 作者: 横内耕;溝口康;岩本英治;麻生久;渡邊敏夫;高須賀晶子;杉本喜憲
• 通讯作者: 杉本喜憲