鸡胚骨形成蛋白（BMPs）基因表达水平的发育性变化

闫成光; 张渝洁; 唐玮琦; 邢晋祎

doi:10.19303/j.issn.1008-0384.2019.11.006

鸡胚骨形成蛋白（BMPs）基因表达水平的发育性变化

doi: 10.19303/j.issn.1008-0384.2019.11.006

1.
临沂大学生命科学学院，山东　临沂　276005
2.
四川农业大学动物科技学院，四川　成都　611130

基金项目: 国家级大学生创新创业训练计划项目（201910452014）；山东省自然科学基金项目（ZR2017LC018）；国家自然科学基金项目（31372333）

详细信息

作者简介:
闫成光（1998−），男，主要从事动物分子遗传研究（E-mail：2521131699@qq.com）

通讯作者:
邢晋祎（1968−），男，博士，教授，主要从事动物分子遗传研究（E-mail：xingjinyi@lyu.edu.cn）

中图分类号: S 814.6
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出版历程
- 收稿日期: 2019-08-13
- 修回日期: 2019-11-13
- 刊出日期: 2019-11-01

Expression of BMPs in Developing Chicken Embryos

1.
College of Life Sciences, Linyi University, Linyi, Shandong　276000, China
2.
College of Animal Sciences and Technology, Sichuan Agricultural University, Chengdu, Sichuan　611130, China

摘要

摘要: 目的分析骨形成蛋白（Bone morphogenetic proteins，BMPs）基因在鸡胚不同发育阶段的表达水平，为进一步研究BMPs基因的功能提供依据。方法选取AA肉鸡种蛋100枚进行孵化，分别于孵化期第1、2、3、4、5、6、9、12、15、18 d（记为E1～E18）选取胚蛋6枚，E1～E6采集整胚，E12～E18分别采集大脑、心脏、肝脏和腿肌组织。采用定量PCR（qPCR）方法检测BMP2、BMP4和BMP7基因的表达丰度。结果 BMP2基因在E1～E6中的表达水平呈现先上升后下降的趋势；BMP2在E4、E5和E6中的表达水平显著高于E1（P＜0.05）；BMP2在大脑中的表达显示，E12显著高于E1（P＜0.05），而到了E18阶段反而极显著低于E1（P＜0.01）；BMP2在心脏和腿肌中的表达均是E9显著或极显著大于E1（P＜0.05或P＜0.01）；鸡胚发育到了后期（E15和E18），BMP2在心脏和肝脏中的表达却显著或极显著低于E1时期（P＜0.05或P＜0.01）。BMP4在E1～E6中的表达水平也是呈现先上升后下降的趋势；BMP4基因在E3～E6整胚和E9胚龄的大脑、心脏、肝脏和腿肌中的表达水平均显著或极显著高于E1胚龄（P＜0.05或P＜0.01）。BMP7在E4、E5和E6的表达与BMP2、BMP4类似，即BMP7基因的表达水平显著或极显著高于E1时期（P＜0.05或P＜0.01），但在E2和E3时期，BMP7表达水平反而降低了，且E2时期极显著低于E1时期（P＜0.01）；在整胚和大脑中的结果显示，E4～E18阶段BMP7基因的表达水平呈现逐渐降低趋势，到E18时期大脑中的BMP7基因的表达水平极显著低于E1时期（P＜0.01）；与E1相比，在心脏中BMP7到E12时期达到最低水平（P＜0.01），而肝脏中BMP7基因表达水平在E12和E15阶段均极显著低于E1（P＜0.01）。结论 BMP2、BMP4和BMP7基因在整胚的表达水平呈现先上升后下降趋势，至E4时期到达最高点；BMP2基因在E9、E12、E15和E18的心脏、肝脏和腿肌中的表达均呈现下降趋势。说明BMPs基因在器官形成初期起着关键作用，之后随着器官发育完成BMPs基因的作用逐渐减弱。
- 肉鸡 /
- 骨形成蛋白 /
- 鸡胚 /
- 表达水平
Abstract: Objective Expression of bone morphogenetic proteins (BMPs) gene in chicken embryos at different developmental stages was studied for further investigation on the functions of the gene. Method One hundred fertilized AA broiler eggs were used for the study. Six eggs were randomly selected each time on the 1^st, 2^nd, 3^rd, 4^th, 5^th, 6^th, 9^th, 12^th, 15^th, and 18^th day (designated as E1 to E18) after hatching. Whole embryos from the eggs of E1 to E6 and the brain, heart, and liver tissues as well as the leg muscles from the embryos of E12 to E18 were collected to determine the expressions of BMP2, BMP4 and BMP7 in the samples by qPCR. Result The BMP2 expressions of E1–E6 increased at first and then decreased. Those of E4, E5, and E6 significantly higher than that of E1 (P＜0.05). BMP2 in the brain of E12 was significantly higher than that of E1 (P＜0.05), but that of E18 became significantly lower than that of E1 (P＜0.01). For E9, the expression of BMP2 in the heart was significantly higher than that for E1 at P＜0.05 and for the leg muscles at P＜0.01. In the later stages of embryonic development, such as E15 and E18, the expressions in the heart and liver were significantly or extremely significantly lower than those of E1 at P＜0.05 or P＜0.01. The expression of BMP4 of E1–E6 also increased initially and followed by a decline. Compared to E1, E3–E6 in embryo and E9 in brain, heart, liver and leg showed significantly higher on BMP4 expressions at P＜0.05 or at P＜0.01. BMP7 of E4 expressed significantly higher than E1 at P＜0.01, and of E5 and E6 at P＜0.05, but lower of E2 or E3 with a statistic significance at P＜0.01 on E2. From E4 through E18, the BMP7 expressions in the embryo and brain declined gradually, and that in the brain reached a significant level at P＜0.01 on E18. In the heart, the lowest expression was observed on E12 (P＜0.01), and in the liver on E12 and E15 (P＜0.01). Conclusion As hatching progressed, the expressions of BMP2, BMP4 and BMP7 in the embryo firstly increased then decreased to arrive at a peak on E4, while BMP2 in the heart, liver and leg decreased on E9, E12, E15 and E18. It indicated that the BMP genes played a crucial role in the early stages of organ formation in fertilized chicken eggs. The effect diminished gradually as the embryonic development came close to completion.
- broiler /
- bone morphogenetic proteins /
- chicken embryo /
- expression

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图 1 BMP2基因的表达水平

注：图1表示BMP2基因在E1～E6整胚和E9～E18胚龄大脑、心脏、肝脏和腿肌中的表达水平（平均值±标准差）（n=6）。*或**表示BMP2基因在E2～E18胚龄中的表达水平与E1中的表达水平相比差异显著（P＜0.05）或极显著（P＜0.01）。

Figure 1. Expression of BMP2

Note: It represent expression levels of BMP2 gene in whole embryos of E1–E6 and in brain, heart, liver and muscle of E9–E18 (Mean ± SD) (n=6), respectively. * or ** indicates significant difference between E1 and E2–E18 (P＜0.05 or P＜0.01).

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图 2 BMP4基因的表达水平

注：图2表示BMP4基因在E1～E6整胚和E9～E18胚龄大脑、心脏、肝脏和腿肌中的表达水平（平均值±标准差）（n=6）。*或**表示BMP4基因在E2～E18胚龄中的表达水平与E1中的表达水平相比差异显著（P＜0.05）或极显著（P＜0.01）。

Figure 2. Expression of BMP4

Note: It represent expression levels of BMP4 gene in whole embryos of E1–E6 and in brain, heart, liver and muscle of E9–E18 (Mean ± SD) (n=6), respectively. * or ** indicates significant difference between E1 and E2–E18 (P＜0.05 or P＜0.01).

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图 3 BMP7基因的表达水平

注：图3表示BMP7基因在E1～E6整胚和E9～E18胚龄大脑、心脏、肝脏和腿肌中的表达水平（平均值±标准差）（n=6）。*或**表示BMP7基因在E2～E18胚龄中的表达水平与E1中的表达水平相比差异显著（P＜0.05）或极显著（P＜0.01）。

Figure 3. Expression of BMP7

Note: It represent expression levels of BMP7 gene in whole embryos of E1–E6 and in brain, heart, liver and muscle of E9–E18 (Mean ± SD) (n=6), respectively. * or ** indicates significant difference between E1 and E2–E18 (P＜0.05 or P＜0.01).

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表 1 qPCR引物序列、扩增片段大小和退火温度

Table 1. Primer sequences, product sizes and annealing temperature applied for qPCR analysis

基因 Gene	引物序列（5′→3′）Primer sequence (5′→3′)	片段大小 Amplicon/bp	退火温度 Annealing temperature /℃	登录号 Accession No.
BMP2	TGGTGGAGGTGGTTCACTT	181	60	NM_204358
BMP2	TTGTGTTTCGCTTGACGC	181	60	NM_204358
BMP4	CTTCGTCTTCAACCTCAGC	150	59	NM_205237
BMP4	ACAGCGGCTTCATCACTT	150	59	NM_205237
BMP7	CTGATTTGTTCCTGCTCG	152	58	AF205877
BMP7	GCTTTGCCCATCTATGCT	152	58	AF205877
GAPDH	CTACACACGGACACTTCAAG	244	59	NM_204305
GAPDH	ACAAACATGGGGGCATCAG	244	59	NM_204305

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