Col2a1基因选择性剪接体在胚胎及出生后不同发育期小鼠眼、耳、软骨中的表达变化

doi:10.6040/j.issn.1673-3770.0.2019.555

摘要/Abstract

摘要： 目的研究Col2a1基因4种选择性剪接体在胚胎及出生后不同发育时期小鼠眼、耳、软骨中的表达变化,为阐明Col2a1在这3种组织或器官发育和功能维持方面的作用提供实验依据。方法取C57bl/6小鼠胚胎12.5 d(E12.5)、16.5 d(E16.5)及出生0d(D0)、7 d(D7)、14 d(D14)、21 d(D21)、28 d(D28)的眼、耳蜗、四肢股关节附近软骨,用real-time PCR检测Col2a1基因中剪接体(IIA、IIB、IIC、IID)的 mRNA在各组织中的表达水平。结果各组织中均检测到Col2a1基因中4种剪接体mRNA的表达;IIA、IIB表达水平较高,IIC、IID表达远低于前二者;在眼、耳中各剪接体表达胚胎期较高,出生至成年后表达逐渐降低,在软骨中除IIB外,各剪接体表达也呈现类似的变化趋势,而IIB 则相反,胚胎期表达较低,随发育时期到成年后表达明显升高;IIA与IIB的比值变化,在软骨中变化极其显著,随发育时期,逐渐出现比例倒置E12.5 IIA表达稍高于IIB(<2倍),到成年期IIB主导表达,IIA几乎检测不到,IIB表达值是IIA的上百倍,而在眼、耳中,IIA/IIB也随发育时期呈下降趋势,但在成年期,IIA表达仍然稍高于IIB。结论 Col2a1基因4种剪接体的表达在眼、耳、软骨中均受发育时期调节,并以IIA和IIB表达为主;IIA在胚胎期中表达旺盛;成年期在软骨中以IIB为主,而在眼、耳组织中,IIA表达仍略高于IIB。

关键词: Col2a1, 选择性剪接体, 发育时期, 眼, 耳, 软骨

Abstract: Objective To study the expression of the alternative splice forms of Col2a1 in mice eyes, ears and cartilage at various development stages and lay a foundation for further understanding the role of Col2a1 in the development and function of these tissues and organs. Methods Obtained eye, ear and cartilage tissues of mice embryos E12.5, E16.5 and postnatal D0, D7, D14, D21, D28, then analyzed the mRNA expression level of the splice forms: IIA, IIB,IIC and IID of Col2a1 through real-time PCR. Results Four splice forms of Col2a1 were detected in three tissues; expression levels of IIA and IIB were higher than IIC and IID, and IIC was the lowest. All splice forms were highiy expressed in eyes and ears at embryonic stages, then decreased at postnatal to adult stages, and splice forms except IIB presented the same trends in cartilage. Expression of IIB was low in the three tissues at the embryonic stage, then its expression increased with development. The ratio of IIA to IIB varied dramatically in cartilage, it was no more than 2 at E12.5, and decreased to 0.004 at D28; while the ratio in eyes and ears also appeared to decline from embryo to adult stages, but was still above 1 at the adult stage. Conclusion Expression of the four splice forms of Col2a1 in eye, ear and cartilage tissue were all regulated by development stages with IIA and IIB being the main forms. IIA expressed actively in all three tissues at embryonic stages, and was replaced by IIB in adult cartilage, while IIA remained higher than IIB in adult eyes and ears.

Key words: Col2a1, Alternative splice form, Development stage, Eye, Ear, Cartilage

中图分类号:

郝佩霞,刘秀珍,张肖林,范义燕,董洪亮,任吉振,尹晶晶,马秀芳. Col2a1基因选择性剪接体在胚胎及出生后不同发育期小鼠眼、耳、软骨中的表达变化[J]. 山东大学耳鼻喉眼学报, 2020, 34(1): 78-84.

HAO Peixia, LIU Xiuzheng, ZHANG Xiaolin, FAN Yiyan, DONG Hongliang, REN Jizhen, YIN Jingjing, MA Xiufang. Expression of the alternative splice forms of Col2a1 in mice eyes, ears and cartilage at various development stages[J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2020, 34(1): 78-84.

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