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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

闫成光,张渝洁,唐玮琦,等. 鸡胚骨形成蛋白(BMPs)基因表达水平的发育性变化 [J]. 福建农业学报,2019,34(11):1276−1282. doi: 10.19303/j.issn.1008-0384.2019.11.006
引用本文: 闫成光,张渝洁,唐玮琦,等. 鸡胚骨形成蛋白(BMPs)基因表达水平的发育性变化 [J]. 福建农业学报,2019,34(11):1276−1282. doi: 10.19303/j.issn.1008-0384.2019.11.006
YAN C G, ZHANG Y J, TANG W Q, et al. Expression of BMPs in Developing Chicken Embryos [J]. Fujian Journal of Agricultural Sciences,2019,34(11):1276−1282. doi: 10.19303/j.issn.1008-0384.2019.11.006
Citation: YAN C G, ZHANG Y J, TANG W Q, et al. Expression of BMPs in Developing Chicken Embryos [J]. Fujian Journal of Agricultural Sciences,2019,34(11):1276−1282. 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
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

Expression of BMPs in Developing Chicken Embryos

  • 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BMP2BMP4BMP7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BMP2BMP4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  BMP2BMP4BMP7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。
  • 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).

    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).

    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).

    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
    TTGTGTTTCGCTTGACGC
    BMP4 CTTCGTCTTCAACCTCAGC 150 59 NM_205237
    ACAGCGGCTTCATCACTT
    BMP7 CTGATTTGTTCCTGCTCG 152 58 AF205877
    GCTTTGCCCATCTATGCT
    GAPDH CTACACACGGACACTTCAAG 244 59 NM_204305
    ACAAACATGGGGGCATCAG
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  • [1] 王广, 李艳, 杨雪松. 鸡胚早期发育过程中细胞迁移的基因调控 [J]. 中国细胞生物学学报, 2011, 33(10):1069−1077.

    WANG G, LI Y, YANG X S. Gene regulation of cell migration during the development of early chick embryo [J]. Chinese Journal of Cell Biology, 2011, 33(10): 1069−1077.(in Chinese)
    [2] HOGAN B L. Bone morphogenetic proteins: multifunctional regulators of vertebrate development [J]. Genes & Development, 1996, 10(13): 1580−1594.
    [3] CHEN D, ZHAO M, MUNDY G R. Bone morphogenetic proteins [J]. Growth Factors, 2004, 22(4): 233−241. doi: 10.1080/08977190412331279890
    [4] EVEN J, ESKANDER M, KANG J. Bone morphogenetic protein in spine surgery: current and future uses [J]. Journal of the American Academy of Orthopaedic Surgeons, 2012, 20(9): 547−52. doi: 10.5435/JAAOS-20-09-547
    [5] ISRAEL D I, NOVE J, KERNS K M, et al. Expression and characterization of bone morphogenetic protein-2 in Chinese hamster ovary cells [J]. Growth Factors, 1992, 7(2): 139−150. doi: 10.3109/08977199209046403
    [6] WOZNEY J M, ROSEN V. Bone morphogenetic protein and bone morphogenetic protein gene family in bone formation and repair [J]. Clinical Orthopaedics and Related Research, 1998, 346: 26−37.
    [7] WOZNEY J M, ROSEN V, CELESTE A J, et al. Novel regulators of bone formation: molecular clones and activities [J]. Science, 1988, 242(4885): 1528−1534. doi: 10.1126/science.3201241
    [8] HE X N, DZIAK R, YUAN X, et al. BMP2 genetically engineered MSCs and EPCs promote vascularized bone regeneration in rat critical-sized calvarial bone defects [J]. PLoS One, 2013, 8(4): e60473. doi: 10.1371/journal.pone.0060473
    [9] LIU Z, YUAN X, FERNANDES G, et al. The combination of nano-calcium sulfate/platelet rich plasma gel scaffold with BMP2 gene-modified mesenchymal stem cells promotes bone regeneration in rat critical-sized calvarial defects [J]. Stem Cell Research & Therapy, 2017, 8(1): 122.
    [10] SHIMASAKI S, ZACHOW R J, LI D, et al. A functional bone morphogenetic protein system in the ovary [J]. Proceedings of the National Academy of Sciences of the United States of America, 1999, 96(13): 7282−7287. doi: 10.1073/pnas.96.13.7282
    [11] SHIMASAKI S, MOORE R K, OTSUKA F, et al. The bone morphogenetic protein system in mammalian reproduction [J]. Endocrine Reviews, 2004, 25(1): 72−101. doi: 10.1210/er.2003-0007
    [12] ROBERT E GODIN, NORMA T TAKAESU, ELIZABETH J ROBERTSON, et al. Regulation of BMP7 expression during kidney development [J]. Development, 1998, 125(17): 3473−3482.
    [13] TSENG Y H, KOKKOTOU E, SCHULZ T J, et al. New role of bone morphogenetic protein 7 in brown adipogenesis and energy expenditure [J]. Nature, 2008, 454(7207): 1000−1004. doi: 10.1038/nature07221
    [14] SAINI S, DURAISAMY A J, BAYEN S, et al. Role of BMP7 in appetite regulation, adipogenesis, and energy expenditure [J]. Endocrine, 2015, 48(2): 405−409. doi: 10.1007/s12020-014-0406-8
    [15] SOMI S, BUFFING A A, MOORMAN A F, et al. Dynamic patterns of expression of BMP isoforms 2, 4, 5, 6, and 7 during chicken heart development [J]. Anatomical Record Part A-Discoveries in Molecular Cellular and Evolutionary Biology, 2004, 279(1): 636−651.
    [16] SCHLUETER J, MÄNNER J, BRAND T. BMP is an important regulator of proepicardial identity in the chick embryo [J]. Developmental Biology, 2006, 295(2): 546−58. doi: 10.1016/j.ydbio.2006.03.036
    [17] GEETHA-LOGANATHAN P, NIMMAGADDA S, HUANG R, et al. Expression pattern of BMPs during chick limb development [J]. Anatomy and Embryology, 2006, 211(S1): 87−93. doi: 10.1007/s00429-006-0129-6
    [18] CHO T J, GERSTENFELD L C, EINHORN T A. Differential temporal expression of members of the transforming growth factor beta superfamily during murine fracture healing [J]. Journal of Bone and Mineral Research, 2002, 17(3): 513−520. doi: 10.1359/jbmr.2002.17.3.513
    [19] KISHIGAMI S, MISHINA Y. BMP signaling and early embryonic patterning [J]. Cytokine & Growth Factor Reviews, 2005, 16(3): 265−278.
    [20] ZHANG H, BRADLEY A. Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development [J]. Development, 1996, 122(10): 2977−2986.
    [21] YU Y H, WILK K, WALDON P L, et al. In vivo identification of Bmp2-correlation networks during fracture healing by means of a limb-specific conditional inactivation of Bmp2 [J]. Bone, 2018, 116: 103−110. doi: 10.1016/j.bone.2018.07.016
    [22] SOUZA C J, CAMPBELL B K, MCNEILLY A S, et al. Effect of bone morphogenetic protein 2(BMP2) on oestradiol and inhibin A production by sheep granulosa cells, and localization of BMP receptors in the ovary by immunohistochemistry [J]. Reproduction, 2002: 363−369. doi: 10.1530/rep.0.1230363
    [23] JUENGEL J L, READER K L, BIBBY A H, et al. The role of bone morphogenetic proteins 2, 4, 6 and 7 during ovarian follicular development in sheep: contrast to rat [J]. Reproduction, 2006, 131: 501−513. doi: 10.1530/rep.1.00958
    [24] ONAGBESAN O M, BRUGGEMAN V, VAN A P, et al. BMPs and BMPRs in chicken ovary and effects of BMP-4 and -7 on granulosa cell proliferation and progesterone production in vitro [J]. American Journal of Physiology and Endocrinological Metabolism, 2003, 285(5): E973−E983. doi: 10.1152/ajpendo.00104.2003
    [25] ZHANG Z, DI R, LIU Q, et al. Expression analysis of five genes in the gonadal axis of Small Tail Han sheep and Sunite sheep [J]. Scientia Agricultura Sinica, 2018, 51: 4710−4719.
    [26] MOWBRAY C, HAMMERSCHMIDT M, WHITFIELD T T. Expression of BMP signalling pathway members in the developing zebrafish inner ear and lateral line [J]. Mechanisms of Development, 2001, 108(1/2): 179−184.
    [27] LA ROSA I, CAMARGO L S, PEREIRA M M, et al. Effects of bone morphogenic protein 4(BMP4) and its inhibitor, Noggin, on in vitro maturation and culture of bovine preimplantation embryos [J]. Reproductive Biology and Endocrinology, 2011, 9(1): 18. doi: 10.1186/1477-7827-9-18
    [28] KIM D, OCÓN-GROVE O, JOHNSON A L. Bone morphogenetic protein 4 supports the initial differentiation of hen (Gallus gallus) granulosa cells [J]. Biology of Reproduction, 2013, 88(6): 161. doi: 10.1095/biolreprod.113.109694
    [29] SU S Y, DONG Z J. Comparative expression analyses of bone morphogenetic protein 4(BMP4) expressions in muscles of Tilapia and common carp indicate that BMP4 plays a role in the intermuscular bone distribution in a dose-dependent manner [J]. Gene Expression Patterns, 2018, 27: 106−113. doi: 10.1016/j.gep.2017.11.005
    [30] YUAN J S, DENG Y, ZHANG Y Y, et al. Bmp4 inhibits goose granulosa cell apoptosis via PI3K/AKT/Caspase-9 signaling pathway [J]. Animal Reproduction Science, 2019, 200: 86−95. doi: 10.1016/j.anireprosci.2018.11.014
    [31] BRAGDON B, MOSEYCHUK O, SALDANHA S, et al. Bone Morphogenetic Proteins: A critical review [J]. Cellular Signalling, 2011, 23(4): 609−620. doi: 10.1016/j.cellsig.2010.10.003
    [32] KUSAKAWA Y, MIKAWA S, SATO K. BMP7 expression in the adult rat brain [J]. IBRO Reports, 2017, 3: 72−86. doi: 10.1016/j.ibror.2017.06.002
    [33] CAMBRIA M T, VILLAGGIO G, FEDERICO C, et al. Bone morphogenic protein BMP7 induces adipocyte differentiation and uncoupling protein UCP1 expression in human bone marrow mesenchymal stem cells [J]. Rendiconti Lincei, 2017, 28(4): 635−641. doi: 10.1007/s12210-017-0643-x
    [34] DONG X G, LI J Y, ZHANG Y Y, et al. Genomic analysis reveals pleiotropic alleles at EDN3 and BMP7 involved in chicken comb color and egg production [J]. Frontiers in Genetics, 2019, 10: 612. doi: 10.3389/fgene.2019.00612
    [35] JIN W Z, TAKAGI T, KANESASHI S N, et al. Schnurri-2 controls BMP-dependent adipogenesis via interaction with smad proteins [J]. Developmental Cell, 2006, 10(4): 461−471. doi: 10.1016/j.devcel.2006.02.016
    [36] RAJESH G, MISHRA S R, PAUL A, et al. Transcriptional and translational abundance of bone morphogenetic protein (BMP) 2, 4, 6, 7 and their receptors BMPR1A, 1B and BMPR2 in buffalo ovarian follicle and the role of BMP4 and BMP7 on estrogen production and survival of cultured granulosa cells [J]. Research in Veterinary Science, 2018, 118: 371−388. doi: 10.1016/j.rvsc.2018.04.002
    [37] ZHANG Y L, LI F Z, FENG X, et al. Genome-wide analysis of DNA Methylation profiles on sheep ovaries associated with prolificacy using whole-genome Bisulfite sequencing [J]. BMC Genomics, 2017, 18: 759. doi: 10.1186/s12864-017-4068-9
    [38] ZHANG Z B, LIU Q Y, DI R, et al. Single nucleotide polymorphisms in BMP2 and BMP7 and the association with litter size in Small Tail Han sheep [J]. Animal Reproduction Science, 2019, 204: 183−192. doi: 10.1016/j.anireprosci.2019.04.001
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  • 收稿日期:  2019-08-13
  • 修回日期:  2019-11-13
  • 刊出日期:  2019-11-01

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