| GnIH基因克隆、表达及对幼龄公兔生殖激素的影响 |
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| 引用本文： | 桑雷,陈冬金,孙世坤,高承芳,王锦祥,陈岩锋,谢喜平. GnIH基因克隆、表达及对幼龄公兔生殖激素的影响[J]. 畜牧兽医学报, 2023, 54(1): 201-212. DOI: 10.11843/j.issn.0366-6964.2023.01.019 |
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| 作者姓名： | 桑雷 陈冬金 孙世坤 高承芳 王锦祥 陈岩锋 谢喜平 |
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| 作者单位： | 1. 福建省农业科学院畜牧兽医研究所, 福州 350013;2. 福建省畜禽遗传育种重点实验室, 福州 350013 |
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| 基金项目： | 国家兔产业技术体系福州综合试验站项目(CARS-43-G-5);福建省自然基金项目(2021J01486);福建省科技计划公益类专项(2021R10260014);福建省农业科学院科技创新团队建设项目(CXTD2021006-2-2) |
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| 摘 要： | 旨在获得兔促性腺激素抑制素(gonadotropin-inhibitory hormone,GnIH)基因,检测其在不同组织和不同发育阶段下丘脑中的表达水平,研究其对幼龄公兔生殖激素分泌的影响。本研究以90日龄健康闽西南黑兔公兔为研究对象,采用RACE技术(rapid-amplification of cDNA ends)从下丘脑组织中克隆出GnIH基因,并对序列进行生物信息学分析;采用实时荧光定量PCR方法(real-time quantitative PCR,qPCR)检测GnIH基因mRNA在90日龄公兔不同组织表达水平(n=5)及11、30、60、90、120、150日龄公兔下丘脑中的表达水平(n=6);连续10 d向80日龄公兔分别注射0、0.5、5和50μg鹌鹑GnIH相关肽(n=10)。第11天早上采集耳静脉血液后,屠宰公兔,采集睾丸组织,分别检测血清中生殖激素浓度(GnRH、FSH、LH、INHB和T)和睾丸组织中睾酮合成相关酶基因(StarR、3β-HSD和p450scc)mRNA的表达水平。结果表明,兔GnIH基因cDNA全长904 bp,包含5′UTR 41 bp...
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| 关 键 词： | GnIH 闽西南黑兔 基因克隆 基因表达 生殖激素 |
| 收稿时间： | 2022-05-05 |
Cloning and Expression of GnIH Gene and Its Effect on Reproductive Hormones Secretion of Young Male Rabbits |
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| SANG Lei,CHEN Dongjin,SUN Shikun,GAO Chengfang,WANG Jinxiang,CHEN Yanfeng,XIE Xiping. Cloning and Expression of GnIH Gene and Its Effect on Reproductive Hormones Secretion of Young Male Rabbits[J]. Chinese Journal of Animal and Veterinary Sciences, 2023, 54(1): 201-212. DOI: 10.11843/j.issn.0366-6964.2023.01.019 |
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| Authors: | SANG Lei CHEN Dongjin SUN Shikun GAO Chengfang WANG Jinxiang CHEN Yanfeng XIE Xiping |
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| Affiliation: | 1. Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China;2. Fujian Key Laboratory of Animal Genetics and Breeding, Fuzhou 350013, China |
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| Abstract: | The purpose of this study was to obtain the sequence of the rabbit GnIH gene, detect its expression patterns in different tissues and different developmental stages in the hypothalamus, and investigate its effect on the secretion of reproductive hormones in young male rabbits. In this study, the 90-day-aged healthy male Minxinan black rabbits were used as experimental animals, the sequence of the GnIH gene was cloned by RACE (rapid-amplification of cDNA ends)and analyzed by bioinformatics methods; the relative expression levels of GnIH in different tissues of male rabbits aged 90 days (n=5) and in the hypothalamus aged 11, 30, 60, 90, 120 and 150 days (n=6) were determined by real-time quantitative PCR (qPCR); the male rabbits aged 80 days were chosen and intraperitoneally injected with 0, 0.5, 5 or 50 μg quail GnIH related peptides for 10 consecutive days, respectively (n=10). These rabbits were slaughtered and their testes were collected after ear vein blood sampling on the morning of the 11th day, and then, the concentrations of reproductive hormones (GnRH, FSH, LH, INHB, and T) in serums and the mRNA levels of testosterone synthesis related enzyme genes (StarR, 3β-HSD and p450scc) in testes were tested, respectively. A total length of 904 bp GnIH gene sequence was cloned successfully, including 41 bp of 5'UTR, 606 bp of CDS, and 227 bp of 3'UTR, encoding 201 amino acids. The GnIH gene expressed in most tissues and its mRNA level was higher in the hypothalamus than in other tissues (P<0.001); Aged from 11 to 150 days, its mRNA levels in hypothalamus reached the lowest point at 90 day-aged (P<0.01), and then significantly increased with increasing age(P<0.001). The young male rabbits were administrated at different doses of GnIH polypeptide. The results showed that the concentration of serum testosterone in group 50 μg was signicantly lower than those in control and 0.5 μg groups(P<0.001), the concentration of testosterone in group 5 μg was significantly lower than those both in control and 0.5 μg groups(P<0.05), and the concentration of LH in group 50 μg was significantly higher than that in control group (P<0.05); the mRNA level of p450scc gene in group 5 μg was significantly higher than those in the other groups(P<0.001), and there were no significant differences in the mRNA levels of other testosterone synthesis related enzyme genes among the GnIH administrating-groups and control group (P>0.05). These results indicated that the GnIH gene might be related to the synthesis and secretion of LH and testosterone, and participate in the regulation of reproductive development of male rabbits, chronic injection with a high dose of GnIH-related peptide leads to drug insensitivity of male rabbits, and the specific regulatory mechanism needs subsequent studies. |
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| Keywords: | GnIH Minxinan black rabbit gene cloning gene expression reproductive hormone |
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