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安哥拉兔(Oryctolagus cuniculus)毛囊发育过程中 Krtap11-1 的特征描述和功能分析。

Characterization and functional analysis of Krtap11-1 during hair follicle development in Angora rabbits (Oryctolagus cuniculus).

机构信息

College of Animal Science and Technology, Yangzhou University, Yangzhou, China.

Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou, China.

出版信息

Genes Genomics. 2020 Nov;42(11):1281-1290. doi: 10.1007/s13258-020-00995-9. Epub 2020 Sep 21.

Abstract

BACKGROUND

Keratin-associated protein (KAP), the structural protein molecule of hair fibers, plays a key role in determining the physical properties of hair. Studies of Krtap11-1 have focused only on its localization. Functional studies of Krtap11-1 in hair follicle development have so far not been reported.

OBJECTIVE

This study aimed to provide evidence for the role of Krtap11-1 in skin and hair development.

METHODS

Full-length cloning and analysis of Krtap11-1 were conducted to ascertain its function. Overexpression vectors and interference sequences were constructed and transfected into RAB-9 cells. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to investigate the hair follicle developmental stage of Krtap11-1, the expression of different tissues, and the effects on other hair follicle development-related genes.

RESULTS

The full length of cloned Krtap11-1 was 947 bp. Krtap11-1 was confirmed to be a hydrophilic protein localized mostly in mitochondria. The greatest mRNA expression was observed in skin. Using a follicle synchronization model, it was found that Krtap11-1 mRNA expression levels first increased then decreased over the passage of time, principally during hair follicle catagen and telogen. Following the overexpression of Krtap11-1, mRNA expression levels of the WNT-2, KRT17, BMP-2, and TGF-β-1 genes increased, and LEF-1 decreased (P < 0.05), the converse after the corresponding use of si-RNA interference.

CONCLUSIONS

Krtap11-1 exerts a promoting effect. The results provide novel insight into the relationship between hair follicle development and Krtap11-1 gene expression.

摘要

背景

角蛋白相关蛋白(KAP)是毛发纤维的结构蛋白分子,在决定毛发的物理性质方面起着关键作用。目前对 Krtap11-1 的研究仅集中在其定位上。关于 Krtap11-1 在毛囊发育中的功能研究尚未见报道。

目的

本研究旨在为 Krtap11-1 在皮肤和毛发发育中的作用提供证据。

方法

通过全长克隆和分析 Krtap11-1 来确定其功能。构建过表达载体和干扰序列,并转染 RAB-9 细胞。采用实时定量聚合酶链反应(qRT-PCR)检测 Krtap11-1 在毛囊发育阶段、不同组织中的表达情况,以及对其他毛囊发育相关基因的影响。

结果

克隆的 Krtap11-1 全长为 947bp。证实 Krtap11-1 是一种主要定位于线粒体的亲水蛋白。在皮肤中观察到最大的 mRNA 表达量。通过毛囊同步化模型发现,Krtap11-1 mRNA 表达水平随时间推移先升高后降低,主要发生在毛囊休止期和退行期。过表达 Krtap11-1 后,WNT-2、KRT17、BMP-2 和 TGF-β-1 基因的 mRNA 表达水平增加,而 LEF-1 减少(P<0.05),相应使用 si-RNA 干扰后则相反。

结论

Krtap11-1 发挥促进作用。研究结果为毛囊发育与 Krtap11-1 基因表达之间的关系提供了新的认识。

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