王 婷 陶亞飛 陳 青 陳 文 黃艷群*
(1.河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院,鄭州450002;2.河南農(nóng)業(yè)大學(xué)國(guó)家家養(yǎng)動(dòng)物種質(zhì)資源平臺(tái),鄭州450002)
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肉雞肥胖基因WDTC1組織表達(dá)特性及油脂類型和水平對(duì)WDTC1基因表達(dá)的影響
王 婷1,2陶亞飛1,2陳 青1,2陳 文1黃艷群1,2*
(1.河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院,鄭州450002;2.河南農(nóng)業(yè)大學(xué)國(guó)家家養(yǎng)動(dòng)物種質(zhì)資源平臺(tái),鄭州450002)
肉雞肥胖基因WDTC1 (WD and tetratricopeptide repeats 1) 是小鼠adipose基因的同源基因,是調(diào)控能量代謝、脂肪沉積和血清甘油三酯水平的關(guān)鍵基因。本試驗(yàn)旨在分析肉雞WDTC1基因組織表達(dá)譜及不同類型和水平的油脂對(duì)肉雞不同組織、不同日齡WDTC1基因表達(dá)的影響。隨機(jī)挑選42日齡商品肉雞母雛5只,進(jìn)行WDTC1基因的組織表達(dá)譜分析。在此基礎(chǔ)上,試驗(yàn)選取240只1日齡科寶肉雞母雛,隨機(jī)分為8個(gè)組(分別在飼糧中添加2.50%、5.00%的亞麻油、玉米油、芝麻油和豬油),每組6個(gè)重復(fù),每個(gè)重復(fù)5只雞,分別于21和42日齡時(shí)從每個(gè)重復(fù)隨機(jī)選取1只肉雞屠宰取樣,通過三因素析因試驗(yàn)設(shè)計(jì)分析油脂類型、水平和肉雞組織以及油脂類型、水平和肉雞日齡對(duì)肉雞WDTC1基因表達(dá)的交互效應(yīng)。結(jié)果表明:1)WDTC1基因在肉雞各個(gè)組織中廣泛表達(dá),其中在卵巢組織中表達(dá)量最高,其次是肝臟、腎臟、腦、肺臟、胸肌、心臟、小腸等組織,在胰腺組織中表達(dá)量最低。2)油脂類型對(duì)42日齡肉雞不同組織中WDTC1基因相對(duì)表達(dá)量的影響極顯著(P<0.01),且油脂類型與肉雞組織(P<0.01)、油脂類型與油脂水平(P<0.05)的交互效應(yīng)顯著;2.50%和5.00%油脂水平時(shí)亞麻油組42日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量顯著高于腹脂組織(P<0.05),且亞麻油組肝臟組織中WDTC1基因的相對(duì)表達(dá)量顯著高于芝麻油組(P<0.05);2.50%油脂水平時(shí)芝麻油組肉雞腹脂組織中WDTC1基因相對(duì)表達(dá)量卻顯著高于亞麻油組(P<0.05);油脂類型和水平對(duì)42日齡肉雞胸肌組織中WDTC1基因相對(duì)表達(dá)量的影響無(wú)顯著差異(P>0.05)。3)油脂類型對(duì)不同日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量的影響極顯著(P<0.01),且油脂類型與油脂水平的交互效應(yīng)顯著(P<0.05);2.50%和5.00%油脂水平時(shí)豬油組21日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量顯著高于42日齡時(shí)(P<0.05);2.50%油脂水平時(shí)亞麻油組肝臟組織中WDTC1基因相對(duì)表達(dá)量顯著高于豬油組(P<0.05),5.00%油脂水平時(shí)亞麻油和玉米油組42日齡肉雞肝臟組織WDTC1基因相對(duì)表達(dá)量顯著高于芝麻油組(P<0.05)。由此可見,肉雞WDTC1基因的表達(dá)有時(shí)空特異性,優(yōu)勢(shì)表達(dá)組織為肝臟,WDTC1基因表達(dá)量受油脂類型、肉雞組織、肉雞日齡、油脂類型與油脂水平以及油脂類型與肉雞組織的交互影響。
肉雞;油脂;WDTC1;基因表達(dá)
Adipose基因,最初在果蠅中被克隆鑒定為肥胖相關(guān)基因并命名為adp[1-3],在哺乳類動(dòng)物中adipose基因的同源基因被命名為WDTC1 (WD and tetratricopeptide repeats 1)基因,該基因從果蠅到人類的保守性均很高[4]。WDTC1蛋白包含WD40重復(fù)和肽重復(fù)序列結(jié)構(gòu)域,它們對(duì)WDTC1蛋白與組蛋白的結(jié)合有重要作用[5-6]。WDTC1蛋白通過與組蛋白2B(H2B)、組蛋白4(H4)和組蛋白去乙酰化酶3形成染色質(zhì)重組復(fù)合體,抑制過氧化物增殖激活受體γ(peroxisome proliferator-activated receptor γ,PPARγ)的轉(zhuǎn)錄活性,進(jìn)而抑制脂肪生成、減少脂肪積累[4,5,7-10]。WDTC1蛋白以劑量依賴方式抑制脂肪的積累,當(dāng)WDTC1基因過表達(dá)時(shí),果蠅體內(nèi)的脂肪沉積減少、血清甘油三酯水平降低;WDTC1基因缺乏時(shí),個(gè)體表現(xiàn)為肥胖[4]。Suh等[10]發(fā)現(xiàn)遺傳型和飼糧誘導(dǎo)肥胖鼠較野生型小鼠和饑餓小鼠脂肪細(xì)胞中WDTC1蛋白水平均有所下降,這表明WDTC1蛋白可能參與調(diào)控哺乳動(dòng)物的脂肪發(fā)生。在人類的研究中也發(fā)現(xiàn)WDTC1基因的變異與肥胖有關(guān),單不飽和脂肪酸攝入量不同對(duì)不同WDTC1基因型個(gè)體的身體質(zhì)量指數(shù)具有不同的效應(yīng)[11]。Wu等[12]發(fā)現(xiàn)了豬WDTC1基因5′側(cè)翼區(qū)有與脂肪細(xì)胞分化及炎癥相關(guān)的關(guān)鍵轉(zhuǎn)錄因子的潛在結(jié)合位點(diǎn),提示W(wǎng)DTC1蛋白在能量代謝上有重要作用。目前有關(guān)WDTC1基因在畜禽上的研究較少,比較基因組學(xué)的分析表明WDTC1基因也是影響雞脂肪沉積的潛在候選基因,但至今尚未見有關(guān)雞WDTC1基因的系統(tǒng)報(bào)道。
玉米油[13]和芝麻油是n-6脂肪酸的代表,含有較高的亞油酸和油酸[14-15];亞麻籽油是n-3脂肪酸的代表,其n-6/n-3多不飽和脂肪酸的比值(約為0.34∶1.00)遠(yuǎn)遠(yuǎn)低于其他油脂[16-17];豬油是富含單不飽和脂肪酸的動(dòng)物油,其油酸含量高達(dá)48.7%[18]。WDTC1基因在動(dòng)物體脂肪沉積及能量代謝中可能具有重要作用,飼糧油脂可能對(duì)其表達(dá)調(diào)控有重要影響。本試驗(yàn)在分析肉雞WDTC1基因組織表達(dá)譜的基礎(chǔ)上,進(jìn)一步研究油脂類型和水平對(duì)其表達(dá)調(diào)控的影響,旨在深入了解雞WDTC1基因的表達(dá)特性。
1.1 試驗(yàn)設(shè)計(jì)
隨機(jī)挑選5只42日齡商品肉雞母雛,以用于雞WDTC1基因的組織表達(dá)譜分析。在此基礎(chǔ)上,采用具有重復(fù)觀測(cè)值的三因素析因試驗(yàn)設(shè)計(jì),探究油脂類型、水平和肉雞組織以及油脂類型、水平和肉雞日齡對(duì)肉雞WDTC1基因表達(dá)的交互效應(yīng)。該研究選用240只1日齡科寶肉雞母雛,隨機(jī)分為8個(gè)組,每組6個(gè)重復(fù),每個(gè)重復(fù)5只雞。試驗(yàn)雞采用籠養(yǎng),每籠5只雞,自由采食和飲水。免疫程序按常規(guī)進(jìn)行。飼糧參照NRC(1994)肉雞營(yíng)養(yǎng)需要配制,分為1~3和4~6周齡2個(gè)階段飼養(yǎng)。1~4組飼糧分別添加2.50%的亞麻油、玉米油、芝麻油和豬油;5~8組飼糧分別添加5.00%的亞麻油、玉米油、芝麻油和豬油。飼糧組成及營(yíng)養(yǎng)水平同張?chǎng)19]文獻(xiàn)中“表1”和“表2”。
1.2 樣品采集
在無(wú)菌操作條件下采集各商品肉雞母雛的腹脂、皮下脂肪、皮膚、腦、心臟、肝臟、脾臟、肺臟、腎臟、卵巢、胸肌、腿肌、肌胃、小腸、胰腺組織以用于WDTC1基因的組織表達(dá)譜分析。在飼糧油脂類型和水平對(duì)肉雞WDTC1基因表達(dá)效應(yīng)的研究中,分別于21日齡和42日齡時(shí)從每個(gè)重復(fù)隨機(jī)挑選1只肉雞屠宰,無(wú)菌操作條件下收集21日齡雞只的肝臟組織以及42日齡雞只的腹脂、肝臟和胸肌組織。所取組織樣品均用焦碳酸二乙酯(DEPC)水清洗后,液氮中速凍,-80 ℃保存?zhèn)溆谩?/p>
1.3 總RNA提取和反轉(zhuǎn)錄反應(yīng)
1.4 引物設(shè)計(jì)
根據(jù)紅色原雞的WDTC1基因序列(GenBank登錄號(hào):XM_417728.4),用Oligo 6.0軟件設(shè)計(jì)熒光定量PCR所用的檢測(cè)引物,并由上海生物工程技術(shù)服務(wù)有限公司合成。正、反向引物及探針如表1。
表1 PCR引物
F:上游 forward;R:下游 reserve;P:探針 probe。
1.5 熒光定量PCR反應(yīng)條件
參照Li等[20]文獻(xiàn),運(yùn)用探針法熒光定量PCR(qPCR)進(jìn)行基因的定量表達(dá)分析。反應(yīng)體系為25.0 μL,包括1.0 μL cDNA(0.1 μL總RNA)、0.5 μL上游引物(12.5 μmol/L)、0.5 μL下游引物(12.5 μmol/L)、2.5 μL TaqManTM探針、0.5 μL RoxⅡ、3.5 μL鎂離子(Mg2+,25 mmol/L)、12.5 μL 2×Mix(包含1.5 mmol/L Mg2+)、4.0 μL超純水;熒光定量PCR反應(yīng)條件為95 ℃預(yù)變性2 min;40個(gè)循環(huán)的95 ℃變性15 s,60 ℃退火1 min。每個(gè)樣品設(shè)置3個(gè)重復(fù)。純化的PCR產(chǎn)物采用微量分光光度計(jì)(NanoDrop 2 000 UV spectrophotometer)測(cè)定濃度后,進(jìn)行梯度(104、105、106、107、108、109和1010)稀釋,用于建立每板的標(biāo)準(zhǔn)曲線。每板均設(shè)陰性對(duì)照。參照Bustin[21]報(bào)道,本研究采用微量定量?jī)x測(cè)定RNA的濃度,反應(yīng)中添加等量的RNA和cDNA進(jìn)行每個(gè)樣品的均一化處理,研究已證明該方法是進(jìn)行基因表達(dá)的定量研究中最可靠的均一化方法。組織表達(dá)圖譜中WDTC1基因的組織相對(duì)表達(dá)量=樣本拷貝數(shù)/胰腺拷貝數(shù)平均值。油脂添加組中WDTC1基因的相對(duì)表達(dá)量=樣本拷貝數(shù)/腹脂平均拷貝數(shù)最小組均值(2.50%油脂水平時(shí)玉米油組均值)。
1.6 數(shù)據(jù)統(tǒng)計(jì)和分析
用SPSS 17.0軟件的獨(dú)立樣本t檢驗(yàn)方法分析雞WDTC1基因的組織表達(dá)譜;用單變量三因素方差完全交互分析方法分析油脂類型、水平和肉雞組織以及油脂類型、水平和肉雞日齡三因素交互作用對(duì)WDTC1基因表達(dá)量的效應(yīng),結(jié)果以平均值(mean)表示。P<0.01為差異極顯著,P<0.05為差異顯著。
2.1 雞WDTC1基因組織表達(dá)譜分析
運(yùn)用qPCR對(duì)雞WDTC1基因在15個(gè)不同組織中的表達(dá)情況進(jìn)行了分析,結(jié)果見圖1。雞WDTC1基因在所有檢測(cè)組織中均有表達(dá),其中在胰腺組織中表達(dá)量最低,在卵巢組織中表達(dá)量最高,為胰腺中表達(dá)量的16.7倍,其次是肝臟、腎臟、腦、肺臟、胸肌、心臟、小腸等組織。除腹脂、皮下脂肪外,其余組織中WDTC1基因的表達(dá)量均顯著高于胰腺中的表達(dá)量(P<0.05)。
2.2 油脂類型和水平對(duì)42日齡肉雞不同組織中WDTC1基因相對(duì)表達(dá)量的影響
油脂類型和水平對(duì)42日齡肉雞不同組織WDTC1基因相對(duì)表達(dá)量的影響見表2。油脂類型對(duì)42日齡肉雞不同組織中WDTC1基因相對(duì)表達(dá)量的影響極顯著(P<0.01),且油脂類型與肉雞組織(P<0.01)、油脂類型與油脂水平(P<0.05)的交互效應(yīng)顯著。
與組織表達(dá)譜結(jié)果一致,42日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量高于胸肌和腹脂組織。5.00%油脂水平時(shí)亞麻油、玉米油和豬油組42日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量均顯著高于腹脂組織(P<0.05);2.50%油脂水平時(shí)亞麻油組42日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量顯著高于腹脂組織(P<0.05);2.50%和5.00%油脂水平時(shí)芝麻油組42日齡肉雞肝臟、腿肌和腹脂組織間WDTC1基因的相對(duì)表達(dá)量無(wú)顯著差異(P>0.05)。
1:腹脂 abdominal fat;2:皮下脂肪 subcutaneous fat;3:皮膚 skin;4:腦 brain;5:心臟 heart;6:肝臟 liver;7:脾臟 spleen;8:肺臟 lung;9:腎臟 kidney;10:卵巢 ovary;11:胸肌 breast muscle;12:腿肌 leg muscle;13:肌胃 muscular stomach;14:小腸 small intestine;15:胰腺 pancreas。
數(shù)據(jù)柱標(biāo)注**表示該組織中表達(dá)量極顯著高于胰腺組織中表達(dá)量(P<0.01),標(biāo)注*表示該組織中表達(dá)量顯著高于胰腺組織中表達(dá)量(P<0.05),NS表示該組織中表達(dá)量與胰腺組織中表達(dá)量差異不顯著(P>0.05)。
Data column with ** mean expression in tissue extremely significantly higher than that in pancreas (P<0.01), and with * mean expression in tissue significantly higher than that in pancreas (P<0.05), while with NS mean no significant difference between expression in this tissue and that in pancreas (P>0.05).
圖1 肉雞WDTC1基因的組織表達(dá)譜
Fig.1 Tissue expression profile ofWDTC1 gene in broilers
42日齡肉雞肝臟組織中,2.50%和5.00%油脂水平時(shí)亞麻油組WDTC1基因相對(duì)表達(dá)量均最高,2.50%油脂水平時(shí)亞麻油組WDTC1基因相對(duì)表達(dá)量顯著高于其他油脂組(P<0.05),5.00%油脂水平時(shí)亞麻油組WDTC1基因相對(duì)表達(dá)量顯著高于芝麻油組(P<0.05);42日齡肉雞腹脂組織中,2.50%油脂水平時(shí)芝麻油和豬油組WDTC1基因的相對(duì)表達(dá)量顯著高于亞麻油組和玉米油組(P<0.05),而5.00%油脂水平時(shí)各油脂組間WDTC1基因的相對(duì)表達(dá)量無(wú)顯著差異(P>0.05)。油脂類型和水平對(duì)肉雞胸肌組織中WDTC1基因的相對(duì)表達(dá)量無(wú)顯著影響(P>0.05);
表2 油脂類型和水平對(duì)42日齡肉雞不同組織中WDTC1基因相對(duì)表達(dá)量的交互效應(yīng)
續(xù)表2項(xiàng)目Items油脂類型Oiltype油脂水平Oillevel/%肝臟Liver胸肌Breastmuscle腹脂Abdominalfat顯著性Significance油脂類型Oiltype## 亞麻油>玉米油≥豬油>芝麻油油脂水平OillevelNS肉雞組織Tissueofbroilers## 肝臟>胸肌>腹脂油脂類型×油脂水平Oiltype×oillevel#油脂類型×肉雞組織Oiltype×tissueofbroilers##油脂水平×肉雞組織Oillevel×tissueofbroilersNS油脂類型×油脂水平×肉雞組織Oiltype×oillevel×tissueofbroilersNS
同列數(shù)據(jù)同一油脂水平肩標(biāo)不同小寫字母表示差異顯著(P<0.05)。同行數(shù)據(jù)肩標(biāo)不同大寫字母表示差異顯著(P<0.05)。##表示差異極顯著(P<0.01),#表示差異顯著(P<0.05),NS表示差異不顯著(P>0.05)。下表同。
Values in the same column of the same oil level with different small letter superscripts mean significant difference (P<0.05). Values in the same row with different large letter superscripts mean significant difference (P<0.05). ## mean extremely significant difference (P<0.01), and # mean significant difference (P<0.05), while NS mean no significant difference (P>0.05). The same as below.
2.3 油脂類型和水平對(duì)不同日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量的影響
油脂類型和水平對(duì)不同日齡肉雞肝臟組織WDTC1基因相對(duì)表達(dá)量的影響見表3。油脂類型對(duì)不同日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量的影響極顯著(P<0.01),且油脂類型與油脂水平的交互效應(yīng)顯著(P<0.05)。
各油脂水平時(shí)各油脂組21日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量均高于42日齡時(shí)。2.50%油脂水平時(shí)玉米油、芝麻油和豬油組21日齡肉雞肝臟組織中WDTC1基因相對(duì)表達(dá)量極顯著高于42日齡時(shí)(P<0.01);5.00%油脂水平時(shí)豬油組肉雞21日齡豬肝臟WDTC1基因相對(duì)表達(dá)量顯著高于42日齡時(shí)(P<0.05);各油脂水平時(shí)亞麻油組21日齡和42日齡肉雞WDTC1基因相對(duì)表達(dá)量無(wú)顯著差異(P>0.05)。
21日齡時(shí),2.50%油脂水平時(shí)豬油組肉雞肝臟組織WDTC1基因相對(duì)表達(dá)量最低,顯著低于亞麻油組(P<0.05),而5.00%油脂水平時(shí)各油脂組肝臟組織WDTC1基因相對(duì)表達(dá)量無(wú)顯著差異(P>0.05);42日齡時(shí),2.50%油脂水平時(shí)亞麻油組肝臟組織WDTC1基因相對(duì)表達(dá)量顯著高于其他油脂組(P<0.05);5.00%油脂水平時(shí)亞麻油和玉米油組肝臟組織WDTC1基因相對(duì)表達(dá)量顯著高于芝麻油組(P<0.05)。
組織表達(dá)譜分析是有效研究基因功能的基礎(chǔ)工作,為進(jìn)一步了解基因的表達(dá)提供有用的參考信息[22]。與果蠅[4]、鼠[10]和豬[12]上的研究結(jié)果一致,WDTC1基因在肉雞組織中廣泛表達(dá),在生殖器官(卵巢)中的表達(dá)量遠(yuǎn)高于其他組織。WDTC1基因在不同物種間的表達(dá)也有所不同,豬WDTC1基因在腎臟、肺臟和脂肪組織中表達(dá)量高,在骨骼肌組織中的表達(dá)量相對(duì)較低[12],而雞WDTC1基因在肝臟、胸肌、腿肌組織中大量表達(dá),在腹脂組織中的表達(dá)量較低,提示W(wǎng)DTC1基因在不同物種間功能的差異性。
肝臟組織中WDTC1基因的表達(dá)呈現(xiàn)出隨日齡的增長(zhǎng)而下降的趨勢(shì),提示了雞WDTC1基因表達(dá)的時(shí)空特異性。亞麻油組肉雞WDTC1基因呈現(xiàn)出在不同油脂水平、不同日齡時(shí)在肝臟組織中的相對(duì)高水平穩(wěn)定表達(dá),提示亞麻酸以非劑量依賴的方式保持WDTC1基因在組織中的穩(wěn)定表達(dá),延緩肝臟WDTC1基因的表達(dá)隨日齡的增長(zhǎng)而降低的趨勢(shì)。
研究表明,長(zhǎng)鏈脂肪酸可以調(diào)控脂代謝相關(guān)基因的表達(dá)[23-24]。陳行杰[25]研究發(fā)現(xiàn),飼糧中添加魚油降低了豬脂肪組織中肥胖基因(ob)和瘦素(leptin)的長(zhǎng)型受體基因的表達(dá)水平,而豆油提高了脂肪組織中PPARγ基因的表達(dá)水平,并提高了背部脂肪和腹部脂肪組織中類胰島素生長(zhǎng)因子(insulin-like grouth factors-1,IGF-1)基因的表達(dá)水平,降低了內(nèi)臟脂肪組織中IGF-1基因的表達(dá)水平。本試驗(yàn)中肝臟組織中WDTC1基因的表達(dá)量高于腹脂組織,肝臟是肉雞WDTC1基因的優(yōu)勢(shì)表達(dá)組織,這可能與鳥類肝臟的脂肪生成能力顯著高于脂肪組織,是脂肪合成的主要場(chǎng)所有關(guān)[26-30]。2.50%油脂水平時(shí)亞麻油上調(diào)了42日齡肉雞肝臟組織WDTC1基因的表達(dá)量而下調(diào)了腹脂組織WDTC1基因的表達(dá)量,芝麻油則下調(diào)了肝臟組織WDTC1基因的表達(dá)量而上調(diào)了腹脂組織WDTC1的表達(dá)量,提示油脂類型以組織特異性的方式影響42日齡肉雞WDTC1基因的表達(dá)。n-3脂肪酸(亞麻油)提高肝臟組織WDTC1基因的表達(dá)量,而n-6脂肪酸(芝麻油)提高脂肪組織WDTC1基因的表達(dá)量,這可能與芝麻油含有較多的固醇、芝麻酚、芝麻酚林和芝麻素等不皂化物,具有抗氧化性和獨(dú)特的生理作用有關(guān)[14]。
表3 油脂類型和水平對(duì)不同日齡肉雞肝臟中WDTC1基因相對(duì)表達(dá)量的交互影響
本試驗(yàn)發(fā)現(xiàn),隨著油脂水平的提高,油脂類型對(duì)42日齡肉雞腹脂組織WDTC1基因表達(dá)量影響的差異進(jìn)一步減?。辉?1日齡和42日齡肉雞肝臟組織中也呈現(xiàn)出類似的現(xiàn)象,隨油脂水平的提高,WDTC1基因的表達(dá)量在不同油脂類型間的差異減小,提示油脂類型以油脂劑量依賴的方式影響WDTC1基因的表達(dá)。
肉雞WDTC1基因的表達(dá)有時(shí)空特異性,其優(yōu)勢(shì)表達(dá)組織為肝臟,在肝臟中的表達(dá)量高于在腹脂中的表達(dá)量,肉雞肝臟組織中WDTC1基因的表達(dá)量隨著肉雞日齡的增長(zhǎng)而降低。油脂類型影響WDTC1基因的表達(dá)量并呈現(xiàn)出與油脂水平、肉雞組織的交互效應(yīng),亞麻油可使肉雞肝臟組織中WDTC1基因的表達(dá)量在不同油脂水平和肉雞日齡均維持較高的水平。油脂類型和水平對(duì)42日齡肉雞胸肌組織中WDTC1的表達(dá)量無(wú)顯著影響。
致謝:
感謝河南農(nóng)業(yè)大學(xué)牧醫(yī)工程學(xué)院王志祥教授對(duì)該文稿所提出的寶貴意見。
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*Corresponding author, professor, E-mail: hyanqun@aliyun.com
(責(zé)任編輯 李慧英)
Tissue Expression Characteristics of Obesity GeneWDTC1 of Broilers and Effects of Oil Type and Level onWDTC1 Gene Expression
WANG Ting1,2TAO Yafei1,2CHEN Qing1,2CHEN Wen1HUANG Yanqun1,2*
(1. Animal Husbandry Medical College of Engineering, Henan Agricultural University, Zhengzhou 450002, China; 2. Henan Agricultural University National Germplasm Resources Platform for Animals, Zhengzhou 450002, China)
Broiler obesity geneWDTC1 (WD and tetratricopeptide repeats 1), a homologous gene of mouseadiposegene, has become a key gene in regulating energy metabolism, fat deposition and serum triglyceride level. This experiment was conducted to analyze the tissue expression profile ofWDTC1 gene in broilers and the effects of different types and different levels of oil onWDTC1 gene expression of broilers in different tissues and at different days of age. Five commercial female broilers at 42 days of age were randomly selected to analysis the tissue expression profile ofWDTC1 gene in broilers. On this basis, two hundred and forty 1-day-old Cobb female broilers were randomly assigned to 8 groups (fed basal diets added with 2.50% and 5.00% linseed oil, corn oil, sesame oil and lard oil, respectively) with 6 replicates per group and 5 broilers per replicate. One broiler of each replicate was selected to slaughter at age of 21 days and 42 days, respectively. The interactive effects of oil type, oil level and tissue of broilers and oil type, oil level and days of age of broilers onWDTC1 gene of broilers were designed to analysis by three factors factorial experiment design. The results showed as follows: 1)WDTC1 gene was found to be widely expressed in various tissues of broilers, and the expression level was the highest in the ovary, then in liver, kidney, brain, lung, breast muscle, heart and small intestine, and the expression level was the lowest in pancreas. 2) The relative expression level ofWDTC1 gene in different tissues of broilers at 42 days of age was significantly affected by the oil type (P<0.01), and the interactive effects of oil type and tissue of broilers (P<0.01) and oil type and oil level (P<0.05) also had significant effects on the relative expression level ofWDTC1 gene. The relative expression level ofWDTC1 gene in liver of broilers at 42 days of age in linseed oil group at 2.50% and 5.00% oil levels was significantly higher than that in abdominal fat (P<0.05), and the relative expression level ofWDTC1 gene in liver in linseed oil group was significantly higher than that in sesame oil group (P<0.05). The relative expression level ofWDTC1 gene in abdominal fat of broilers in sesame oil group at 2.50% oil level was significantly higher than that in linseed oil group (P<0.05). Oil type and level had no significant effect on the relative expression level ofWDTC1 gene in breast muscle of broilers at 42 days of age (P>0.05). 3) The relative expression level ofWDTC1 gene in liver of broilers at different days of age was significantly affected by the oil type (P<0.01), and the interactive effect of oil type and oil level also had significant effect on the relative expression level ofWDTC1 gene (P<0.05). The relative expression level ofWDTC1 gene in liver of broilers at 21 days of age in lard oil group at 2.50% and 5.00% oil levels was significantly higher than that at 42 days of age (P<0.05). The relative expression level ofWDTC1 gene in liver in linseed oil group at 2.50% oil level was significantly higher than that in lard oil (P<0.05), and the relative expression level ofWDTC1 gene in liver of broilers at 42 days of age in linseed oil and corn oil groups at 5.00% oil level was significantly higher than that in linseed oil group (P<0.05). In conclusion,WDTC1 gene expression of broilers has the temporal and spatial specificities, and liver is the predominantly expression tissue.WDTC1 expression level is affected by oil type, tissue of broilers, days of age of broilers and the interaction effects of oil type and oil level, oil type and tissue of broilers.[ChineseJournalofAnimalNutrition, 2016, 28(11):3511-3519]
broilers; oil;WDTC1; gene expression
2016-04-25
國(guó)家自然科學(xué)基金(31272434)
王 婷(1992—),女,河南周口人,碩士研究生,動(dòng)物遺傳育種與繁殖專業(yè)。E-mail: wangting917@163.com
*通信作者:黃艷群,教授,碩士生導(dǎo)師,E-mail: hyanqun@aliyun.com
10.3969/j.issn.1006-267x.2016.11.018
S831.5;S816.4
A
1006-267X(2016)11-3511-09
動(dòng)物營(yíng)養(yǎng)學(xué)報(bào)2016年11期