突触融合蛋白4在不同毛色小鼠皮肤组织的差异表达

doi:10.11843/j.issn.0366-6964.2015.06.011

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (6): 957-964.doi: 10.11843/j.issn.0366-6964.2015.06.011

突触融合蛋白4在不同毛色小鼠皮肤组织的差异表达

谢建山^1#，张丹瑾^1#，郭艳妮¹，范瑞文¹，许冬梅¹，杨玉静¹，聂瑞强¹，于秀菊¹，段力升¹，朱芷葳²，高文俊¹，赫晓燕¹，董常生^1*

（1.山西农业大学动物科技学院，太谷 030801；2.山西农业大学生命科学学院，太谷 030801）

收稿日期:2015-02-09 出版日期:2015-06-23 发布日期:2015-06-19
通讯作者: 董常生，教授，博士生导师，E-mail:cs_dong@sxau.edu.cn
作者简介:谢建山（1981-），男，山西朔州人，博士生，主要从事动物毛色形成机理研究，E-mail:xiejs@live.com；张丹瑾（1989-），女，山西太原人，硕士，主要从事羊驼毛色的研究，E-mail: ellen7789@163.com；谢建山与张丹瑾为并列第一作者
基金资助:
国家“863”计划（2013AA102506）；公益性行业（农业）科研专项（201303119）；农业部“948”项目（2-11-Z33）；肉牛、肉羊标准化生产体系的建立-子项目-利用转microRNA技术培育彩色转基因细毛羊；山西省重点学科建设经费；山西省基础研究项目（青年科技研究基金）（2014021028-2）

Differential Expression of the Syntaxin 4 in Differential Coat Color Mice Skin

XIE Jian-shan^1# ，ZHANG Dan-jin^1# ，GUO Yan-ni¹，FAN Rui-wen¹，XU Dong-mei¹，YANG Yu-jing¹，NIE Rui-qiang¹，YU Xiu-ju¹，DUAN Li-sheng¹，ZHU Zhi-wei²，GAO Wen-jun¹ ，HE Xiao-yan¹，DONG Chang-sheng^1*

(1.College of Animal Science and Veterinary Medicine，Shanxi Agricultural University，Taigu 030801，China；2.College of Life Science，Shanxi Agricultural University，Taigu 030801，China)

Received:2015-02-09 Online:2015-06-23 Published:2015-06-19

摘要/Abstract

摘要：

旨在探讨突触融合蛋白4（Syntaxin4，STX4）在皮肤组织细胞中的表达与定位，确定其是否与毛色形成存在相关性。选择白、灰、黑3种毛色皮肤组织样品（6只昆明小鼠白毛色背部皮肤、6只C57BL/6小鼠灰毛色腹部皮肤和黑毛色背部皮肤）和体外培养小鼠黑色素细胞样品，通过PCR扩增、Real-time PCR、免疫组化和Western blot 技术对STX4的表达情况进行定性和定量分析。结果表明：在3种毛色皮肤样品和黑色素细胞样品中均扩增出897 bp CDS区序列片段；Real-time PCR检测显示，STX4在白灰黑3种毛色皮肤样品中均有表达，黑色皮肤表达量最高，灰色皮肤表达量次之，分别是白色皮肤的3.44倍和1.92倍；免疫组化结果表明，在白色和黑色皮肤表达于整个毛囊, 包括角化细胞；在体外培养黑色素细胞也显示阳性表达； Western blot结果显示，在白、灰、黑色皮肤和黑色素细胞样品中均有STX4阳性条带，表达量与荧光定量结果一致。综上表明，STX4在小鼠皮肤组织、毛囊角化细胞、黑色素细胞有效表达，且表达量在白灰黑皮肤中呈递增趋势，推测STX4与小鼠毛色形成呈正相关性。

关键词: 突触融合蛋白4, 差异表达, 小鼠, 不同毛色, 毛囊

Abstract:

The aim of the present study was to determine whether STX4 is associated with coat color formation in mice. The expression and localization of STX4 in mice skins and melanocytes were investigated. Three different coat color skins, white, grey and black, were collected from the back of 6 white Kunming mice, the abdomen of 6 C57BL/6 mice and the back of 6 C57BL/6 mice, respectively. Melanocytes were cultured in vitro. The expression of STX4 in these skin and cell samples were analyzed by standard RT-PCR, quantitative real-time PCR, immunohistochemical staining and Western blot. The 897 bp sequence of STX4 CDS region was successfully amplified from 3 different coat color mice skins and melanocytes cultured in vitro by RT-PCR. Real-time PCR analysis revealed that STX4 was expressed in mice skins of all coat colors. STX4 showed the highest expression in black skin, which was 3.44 times higher than white skin. The expression of STX4 in grey skin was 1.92 times higher than white skin. Immunohistochemical analysis showed that STX4 was expressed in whole hair follicles in white and black skins including keratinocytes, as well as in melanocytes cultured in vitro. Western blot results showed positive STX4 bands in white, grey and black skin samples and melanocytes cultured in vitro. These results were consistent with the real-time PCR results. In conclusion, STX4 is expressed in mice skins, hair follicles, keratinocytes, and melanocytes, and the expression of STX4 increases as the coat color deepens. Therefore, it is speculated that STX4 is positively correlated to coat color formation in mice.

Key words: syntaxin 4（STX4）, differential expression, mouse, different coat color, hair follicle

中图分类号:

S852

谢建山，张丹瑾，郭艳妮，范瑞文，许冬梅，杨玉静，聂瑞强，于秀菊，段力升，朱芷葳，高文俊，赫晓燕，董常生. 突触融合蛋白4在不同毛色小鼠皮肤组织的差异表达[J]. 畜牧兽医学报, 2015, 46(6): 957-964.

XIE Jian-shan，ZHANG Dan-jin，GUO Yan-ni，FAN Rui-wen，XU Dong-mei，YANG Yu-jing，NIE Rui-qiang，YU Xiu-ju，DUAN Li-sheng，ZHU Zhi-wei，GAO Wen-jun,HE Xiao-yan，DONG Chang-sheng. Differential Expression of the Syntaxin 4 in Differential Coat Color Mice Skin[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(6): 957-964.

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突触融合蛋白4在不同毛色小鼠皮肤组织的差异表达

Differential Expression of the Syntaxin 4 in Differential Coat Color Mice Skin

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