畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (3): 454-461.doi: 10.11843/j.issn.0366-6964.2017.03.008

• 生物技术与繁殖 • 上一篇    下一篇

公山羊β防御素124的表达谱分析及其在繁殖器官中的定位

杜海燕,邰苗苗,姜玉锁,任有蛇,靳黎,乔利英,张春香*   

  1. (山西农业大学动物科技学院,太谷 030801)
  • 收稿日期:2016-10-19 出版日期:2017-03-23 发布日期:2017-03-23
  • 通讯作者: 张春香,E-mail:zhchx66@126.com
  • 作者简介:杜海燕(1977-),女,山西偏关人,博士生,讲师,主要从事动物遗传育种与繁殖研究,E-mail:dhy786@126.com
  • 基金资助:

    国家自然科学基金(31572407)

Expression Pattern Analysis of Goat Beta-defensin 124 and Its Location in the Reproductive Organs

DU Hai-yan, TAI Miao-miao, JIANG Yu-suo, REN You-she, JIN Li, QIAO Li-ying, ZHANG Chun-xiang*   

  1. (College of Animal Science and Veteriary Medicine, Shanxi Agricultural University, Taigu 030801,China)
  • Received:2016-10-19 Online:2017-03-23 Published:2017-03-23

摘要:

旨在研究公山羊β防御素124(Goat beta-defensin 124, gDB 124)的表达谱及其蛋白在繁殖器官中的分布特点。采集1周龄、2~6月龄公羊各3只(体重相近)睾丸和附睾,以及18月龄公山羊睾丸、附睾头、附睾体、附睾尾、心、肝、脾、肺、肾和背最长肌组织。用qRT-PCR分析gDB 124 mRNA的表达情况;采用免疫荧光技术分析睾丸、附睾头、附睾体和附睾尾gDB 124分布特点。结果显示:除背最长肌中无表达外,gDB 124 mRNA在18月龄公山羊睾丸、附睾头、附睾体、附睾尾、心、肝、脾、肺和肾组织均有表达,且附睾头表达量极显著高于其他组织(P<0.01);睾丸gDB 124 mRNA表达量6月龄时显著高于其他月龄;附睾头、附睾体和附睾尾gDB 124 mRNA表达量随月龄的增加呈上升趋势;1周龄公山羊睾丸和附睾gDB 124 mRNA表达量不存在显著差异,3、4、5、6、18月龄时附睾头gDB 124 mRNA 表达量显著高于睾丸、附睾体和附睾尾(P<0.05);免疫荧光定位技术分析显示:gDB 124在公山羊睾丸和附睾中均有分布,但主要是在附睾头上皮细胞中。根据荧光信号强度gDB 124的表达量顺序:附睾头>附睾体>附睾尾>睾丸,该结果与qRT-PCR分析结果相一致。公山羊gDB 124 mRNA表达具有明显组织特异性和时间依赖性。

Abstract:

To study the expression pattern of goat beta-defensin 124 (gDB 124) in different organs and the location of gDB 124 in testis and epididymis of adult bucks. Testis, caput epididymidis, corpus epididymidis and cauda epididymidis were collected from 1-week-old male kids, 2-6-month-old male kids and 18-month-old bucks. Part of the tissues were gathered from heart, liver, spleen, lung, kindey and longissimus dorsi of bucks at the age of 18 months. Analysis of gDB 124 mRNA expression pattern was detected by qRT-PCR. Location of gDB 124 in the reproductive organs was determined by immunofluorescence. The results of qRT-PCR showed that gDB 124 mRNA was detected in testis, caput, corpus, cauda, heart, liver, spleen, lung and kidney of bucks at the age of 18 months except the longissimus dorsi,and the expression level in caput of epididymis was extremely higher than that in other tissues (P<0.01). Moreover, gDB 124 mRNA expression level in testis of buck at the age of 6 months was significantly higher than those at other ages. And the expressions in caput, corpus, cauda of epididymis were increased with the increase of the age. There was no obvious difference at gDB 124 mRNA expression levels between testis and epididymis of 1-week-old bucks. gDB 124 mRNA expressions in caput of epididymis were extremely higher than those in testis, corpus, cauda of epididymis of bucks at 3-6 and 18 months old (P<0.05).The result of immunofluorescence suggested that gDB 124 protein was detected in testis and epididymis, and mainly located in false stratified columnar epithelium cells of caput of epididymis. The order of the expression level was caput >corpus>cauda> testis, which was consistent with expression pattern of gDB 124 mRNA in testis and epididymis. In conclusion, the expression of gDB 124 was in the spatio-temporal expression.

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