矿化相关基因CA2和CA13在鸡双侧下颌骨髁部不对称导致交叉喙

doi:10.1016/j.jia.2023.04.014

Journal of Integrative Agriculture ›› 2024, Vol. 23 ›› Issue (7): 2379-2390.DOI: 10.1016/j.jia.2023.04.014

矿化相关基因CA2和CA13在鸡双侧下颌骨髁部不对称导致交叉喙

收稿日期:2020-01-13 接受日期:2023-02-23 出版日期:2024-07-20 发布日期:2024-07-02

Asymmetric expression of CA2 and CA13 linked to calcification in the bilateral mandibular condyles cause crossed beaks in chickens

Lei Shi^{1, 2*}, Yanyan Sun^1*, Yunlei Li¹, Hao Bai³, Jingwei Yuan¹, Hui Ma¹, Yuanmei Wang¹, Panlin Wang¹, Aixin Ni¹, Linlin Jiang¹, Pingzhuang Ge¹, Shixiong Bian¹, Yunhe Zong¹, Jinmeng Zhao¹, Adamu M. Isa¹, Hailai H. Tesfay¹, Jilan Chen^1#

¹ State Key Laboratory of Animal Biotech Breeding/Key Laboratory of Animal (Poultry) Genetics Breeding and Reproduction of Ministry of Agriculture and Rural Affairs/Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

² College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China

³ Joint International Research Laboratory of Agriculture and Agri-Product Safety, Ministry of Education/Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China

Received:2020-01-13 Accepted:2023-02-23 Online:2024-07-20 Published:2024-07-02
About author:Lei Shi, E-mail: shilei2017@foxmail.com; Yanyan Sun, E-mail: yanyansun2014@163.com; # Correspondence Jilan Chen, Tel: +86-10-62816005, E-mail: chen.jilan@163.com * These authors contributed equally to this study.
Supported by:
This work was supported by the Beijing Featured Livestock and Poultry Genetic Resources Preservation Project, China (202203310002). China Agriculture Research System of MOF and MARA (CARS40), the Agricultural Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences (ASTIPIAS04), and the Central Guidance on Local Science and Technology Development Fund of Hebei Province, China (236Z6602G).

摘要/Abstract

摘要：

交叉喙是多基因控制的复杂性状，目前已在全球至少12个鸡种中公开报道，发生率介于0.2%~7.4%。交叉喙严重影响个体采食和饮水，降低生产性能，损害动物福利。为探究交叉喙性状分子调控机制，本研究首先分析交叉喙鸡下颌骨形态特征，再通过RNA-seq技术筛选交叉喙鸡与正常鸡双侧下颌骨髁部的差异表达基因，结合功能富集分析鉴定关键候选基因。在此基础上，结合组织表达谱和细胞水平试验，验证关键候选基因导致鸡交叉喙的分子机制。结果表明，交叉喙由左右侧下颌骨长度不等导致，即某一侧下颌骨支短于对侧正常骨支，下喙偏向于下颌骨短支侧。由于下颌骨髁部是下颌骨支的生发中心，通过对交叉喙鸡短骨支侧髁部和正常骨支髁部进行转录组分析，共筛选110个差异表达基因，并显著富集于碳酸酐酶活性条目，差异表达基因为碳酸酐酶2（CA2）和碳酸酐酶13（CA13）。结合组织表达谱分析，发现CA2和CA13高表达于鸡下颌骨髁部(P<0.05)。通过比较成骨细胞增殖期与矿化期的CA2和CA13表达特征，发现两者在矿化期成骨细胞中显著上调。对下喙左偏和下喙右偏的交叉喙个体进行验证，发现短骨支侧髁部的CA2和CA13表达水平低于对侧正常骨支髁部(P<0.05)。为解析CA2和CA13基因功能，本试验在鸡成骨细胞培养体系中添加碳酸酐酶抑制剂，发现碳酸酐酶活性与成骨细胞矿化存在相关；进一步向成骨细胞中转染CA13干扰慢病毒，发现CA13被干扰后引起鸡成骨细胞矿化能力下降。研究结果揭示了CA2和CA13可能是北京油鸡交叉喙性状的潜在调控因子，但在其他品种中仍需进一步验证。此外，考虑到碳酸酐酶编码基因在人类中同样高表达，本研究也可能为人类下颌骨畸形引起的错颌和颅面不对称病因等提供参考。

创新点：本研究基于个体对称部位为研究对象的试验设计，结合转录组分析，揭示了CA2和CA13的不对称表达是鸡交叉喙性状形成的原因，研究结果将为地方鸡种复杂性状的研究提供新思路。

Abstract:

Crossed beak is a complex mode of inheritance with prevalence ranging from 0.2 to 7.4% in at least 12 chicken strains worldwide. To reveal the intrinsic factors causing crossed beaks, genes expression patterns in bilateral mandibular condyle between affected and normal birds were characterized by RNA sequencing analysis in the present studies. Crossed beak was induced by short length of unilateral mandibular ramus, and a total of 110 differentially expressed genes were up- or down-regulated in the affected (short) mandibular condyle side as compared to the normal side. Carbonic anhydrase 2 (CA2) and Carbonic anhydrase 13 (CA13) were enriched in the carbonate dehydratase activity, and high-expressed in mandibular condyle and osteoblasts (P<0.05). However, both were low-expressed in short mandibular condyle side of affected birds (P<0.05). The carbonate dehydratase inhibitor experiments confirmed that there is positive association between the calcification and carbonic anhydrase isoenzymes. Quantitative analysis with cetylpyridinium chloride showed a decrease in calcification when the cells were transfected with an anti-CA13 shRNA. Our research suggested that CA2 and CA13 are down-calcified in shortside mandibular condyle, and caused mandibular ramus to grow slowly. CA2 and CA13 have the critical role in crossed beaks by regulating calcification of mandibular condyle.

Key words: chicken , crossed beak , carbonic anhydrase , calcification , CA2 , CA13

Lei Shi, Yanyan Sun, Yunlei Li, Hao Bai, Jingwei Yuan, Hui Ma, Yuanmei Wang, Panlin Wang, Aixin Ni, Linlin Jiang, Pingzhuang Ge, Shixiong Bian, Yunhe Zong, Jinmeng Zhao, Adamu M. Isa, Hailai H. Tesfay, Jilan Chen. 矿化相关基因CA2和CA13在鸡双侧下颌骨髁部不对称导致交叉喙[J]. Journal of Integrative Agriculture, 2024, 23(7): 2379-2390.

Lei Shi, Yanyan Sun, Yunlei Li, Hao Bai, Jingwei Yuan, Hui Ma, Yuanmei Wang, Panlin Wang, Aixin Ni, Linlin Jiang, Pingzhuang Ge, Shixiong Bian, Yunhe Zong, Jinmeng Zhao, Adamu M. Isa, Hailai H. Tesfay, Jilan Chen. Asymmetric expression of CA2 and CA13 linked to calcification in the bilateral mandibular condyles cause crossed beaks in chickens[J]. Journal of Integrative Agriculture, 2024, 23(7): 2379-2390.

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矿化相关基因CA2和CA13在鸡双侧下颌骨髁部不对称导致交叉喙

Asymmetric expression of CA2 and CA13 linked to calcification in the bilateral mandibular condyles cause crossed beaks in chickens

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