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农业生物技术学报  2019, Vol. 27 Issue (10): 1869-1877    DOI: 10.3969/j.issn.1674-7968.2019.10.017
  专题:高原特色动物生理生化研究 本期目录 | 过刊浏览 | 高级检索 |
CRY1在雄性牦牛生殖轴中的表达与分布
陈健楠1, 王琪2, 王媛媛2, 吕琛2, 赵彩英1, 张勇1,2, 马友记3, 张全伟1,*, 赵兴绪1,2,*
1 甘肃农业大学 生命科学技术学院,兰州 730070;
2 甘肃农业大学 动物医学院,兰州 730070;
3 甘肃农业大学 动物科学技术学院,兰州 730070
Expression and Distribution of CRY1 in the Reproductive Axis of Male Yak (Bos grunniens)
CHEN Jian-Nan1, WANG Qi2, WANG Yuan-Yuan2, LV Chen2, ZHAO Cai-Ying1, ZHANG Yong1,2, MA You-Ji3, ZHANG Quan-Wei1,*, ZHAO Xing-Xu1,2,*
1 College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
3 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
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摘要 隐花色素1 (cryptochrome 1, CRY1)基因是哺乳动物生物钟基因之一,其表达蛋白既是光受体又是生物钟的组成部分,经体液和神经途径送达效应器,调节生理、生化和节律行为。本研究采集牦牛(Bos grunniens)的下丘脑、垂体、松果体及睾丸组织,通过反转录聚合酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)技术对CRY1编码区序列进行克隆和序列分析,通过荧光定量PCR (real-time fluorescent quantitative PCR, qRT-PCR)、Western blot和免疫组织化学染色技术分析CRY1在成年雄性牦牛生殖轴系及不同年龄段(30日龄, 2, 4和6岁龄)雄性牦牛睾丸中的表达规律。结果显示,CRY1基因CDS全长为1 712 bp,ORF为1 257 bp (GenBank No. MN460656),共编码418个氨基酸残基,无跨膜结构区,编码的蛋白属可溶性蛋白;组织表达与分布结果发现,CRY1 mRNA和蛋白在雄性牦牛下丘脑、垂体、睾丸及松果体组织中均有表达,在松果体组织中的表达水平极显著高于其他组织(P<0.01);免疫组织化学染色结果显示,在下丘脑室旁核细胞、垂体细胞、睾丸各级生精细胞、松果体细胞和神经胶质细胞上均有阳性表达;不同年龄段睾丸组织中表达结果发现,CRY1 mRNA和蛋白在30日龄睾丸中的表达量最低,随着年龄的增长,表达量逐渐增高。研究结果表明,CRY1在动物进化中比较保守,在雄性牦牛性腺轴和不同年龄牦牛睾丸中均有表达,推测其可能在牦牛生殖过程中起着重要作用。上述结果为牦牛生物节律基因CRY1的生物学功能及其在牦牛生殖生理调控的相关研究提供依据。
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陈健楠
王琪
王媛媛
吕琛
赵彩英
张勇
马友记
张全伟
赵兴绪
关键词 牦牛隐花色素1基因(CRY1)生殖轴基因克隆表达分布    
Abstract:Cryptochrome 1 (CRY1) gene is a member of mammalian clock genes. Its expressed protein is both a photoreceptor and a part of the circadian clock, which could be delivered to the effector via the humoral and neural pathways to regulate mammalian physiology, biochemistry and behavior. In this study, the samples (the hypothalamus, pituitary, pineal and testicular tissues) of yak were collected, and the sequence of CRY1 coding region was cloned and the expression of CRY1 in adult male yak reproductive axis and yak testes of different ages (30 d, 2, 4 and 6 years old) was analyzed by qRT-PCR (real-time fluorescent quantitative PCR), Western blot and immunohistochemical staining. The results showed that the CRY1 gene CDS was 1 712 bp in length and the ORF was 1 257 bp (GenBank No. MN460656), encoding a total of 418 amino acid residues, without transmembrane domain, and the encoded protein was a soluble protein. Tissue expression and localization revealed that CRY1 mRNA and protein were expressed in the hypothalamus, pituitary, testis and pineal tissues of yak, and the expression level in pineal tissue was significantly higher than other tissues (P<0.01). Immunohistochemical staining showed positive expression in the hypothalamic paraventricular nucleus, pituitary cells, testicular spermatogenic cells, pineal cells and glial cells. The expression of CRY1 mRNA and protein in the testis of 30 d was the lowest in the testis tissues of different age groups, and the expression level gradually increased with age. The results showed that the CRY1 gene was relatively more conservative in animal evolution and was expressed in the male yak gland axis and the yak testes of different ages, suggesting that it might play an important role in the reproductive process of yak. The above results provides the basis for the biological function of the yak biorhythm gene CRY1 and its related research on the regulation of yak reproductive physiology.
Key wordsYak    Cryptochrome 1 gene (CRY1)    Reproductive axis    Gene cloning    Expression and distribution
收稿日期: 2019-03-20     
ZTFLH:  S824  
基金资助:国家高技术研究发展计划(863)项目(No. 2013AA102505-3)和甘肃农业大学学科建设基金(No. GSAU-XKJS-2018—160)
通讯作者: zhangqw@gsau.edu.cn;zhaoxx@gsau.edu.cn   
引用本文:   
陈健楠, 王琪, 王媛媛, 吕琛, 赵彩英, 张勇, 马友记, 张全伟, 赵兴绪. CRY1在雄性牦牛生殖轴中的表达与分布[J]. 农业生物技术学报, 2019, 27(10): 1869-1877.
CHEN Jian-Nan, WANG Qi, WANG Yuan-Yuan, LV Chen, ZHAO Cai-Ying, ZHANG Yong, MA You-Ji, ZHANG Quan-Wei, ZHAO Xing-Xu. Expression and Distribution of CRY1 in the Reproductive Axis of Male Yak (Bos grunniens). 农业生物技术学报, 2019, 27(10): 1869-1877.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.10.017     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I10/1869
 
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