绒山羊皮肤毛囊中生物钟基因表达的研究

发布时间：2018-03-15 11:24

本文选题：绒山羊　切入点：初级毛囊　出处：《内蒙古农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：山羊绒毛生长发育呈明显的季节性变化,它的皮肤作为感知外环境温度变化和水分缺失的交界面,能够使机体内环境与外环境相协同发展。这表明绒山羊皮肤中可能存在着一个时间调控机制,而这一时间机制受下丘脑视交叉上核(SCN)上生物钟基因的调控。因此,生物钟基因之间相互作用的关系影响着皮肤毛囊的生长发育。本研究为进一步证实生物钟基因是否形成一个近乎以24小时为周期的转录翻译反馈循环系统来调控皮肤毛囊的生长发育,以13个生物钟基因CSNKIE、CENK1D、TIM、CLOCK、 PER1、PER2、PER3、TIM、CRY1、CRY2、BHLHE40、BHLHE41、ARNTL、NR1D1为研究对象,采用RNA-Seq技术初步分析不同生长时期初级毛囊和次级毛囊上生物钟基因的表达模式；此外,为深入探究CLCOK、TIM, PER1、CRY1基因在皮肤次级毛囊中的相互作用及其分子机制,文章进而应用荧光定量PCR技术对以上4个钟基因在次级毛囊上的表达做了定量分析。通过以上研究分析,本文得到如下结果：1、CSNKIE基因在皮肤毛囊中进行高表达,其表达量高出其它钟基因几十倍。生物钟基因在山羊皮肤初级毛囊上的表达量依次为：CSNKIE、CSNKID、BHLHE40、ARNTL CRY1 PER1、CLOCK、CRY2、NR1D1 PER2 PER3、BHLHE41;皮肤次级毛囊上的表达量按从大到小的顺序依次为：CSNKIE CSNKID、ARNTL、BHLHE40、NR1D1、PER1、 CRY1、CRY2、BHLHE41、CLOCK、PER2 PER3。2、CSNKID、CLOCK、CRY2、PER3和BHLHE41基因在绒山羊皮肤毛囊上的表达随毛囊生长时期和采样时间的变化存在显著差异；NR1D1基因在初级毛囊同一生长时期不同采样时间的表达差异显著,ARNTL和PER2基因随次级毛囊生长时期的不同,其表达量有显著差异。3、生物钟基因在绒山羊皮肤次级毛囊中的表达量(荧光定量PCR)依次为：CLOCK TIM PER1 CRY1。4、皮肤次级毛囊兴盛期,CLOCK基因、TIM基因、PER1基因、CRYl基因的高振幅节律表达分别于04：00、08：00、08：00、16：00进行“报时”；皮肤次级毛囊休止期,这四个钟基因的高振幅节律表达分别于08：00、04：00、04：00、16：00进行“报时”。
[Abstract]:The growth and development of cashmere wool show obvious seasonal changes. Its skin acts as the interface between the temperature change of the outer environment and the water loss. This suggests that there may be a time regulation mechanism in the skin of cashmere goats, which is regulated by the biological clock gene on the supraoptic nucleus of hypothalamus SCN. The interaction between the biological clock genes affects the growth and development of the skin hair follicles. This study is intended to further confirm whether the biological clock gene is regulated by a nearly 24-hour transcriptional translation feedback circulatory system. The growth and development of skin follicles, The expression patterns of clock genes in primary hair follicles and secondary hair follicles of 13 biological clock genes, CSNKIEX CENK1D, PER1PER2PER3, CRY2, BHLHE40, BHLHE41ANTLNR1D1, were preliminarily analyzed by RNA-Seq technique, and the expression patterns of clock genes in primary hair follicles and secondary hair follicles at different growth stages were analyzed by RNA-Seq technique. In order to explore the interaction of CRY1 gene and its molecular mechanism in the skin secondary follicles, the expression of the four clock genes in the secondary hair follicles was quantitatively analyzed by fluorescence quantitative PCR. The following results were obtained: 1: 1 CSNKIE gene was overexpressed in the skin follicles. The expression of biological clock gene in the primary hair follicles of goats was in the order of: CSNKIEI BHLHE40ARNTL CRY1 PER1 CLOCK1 PER2 PER3HEL411.The order of expression on the skin secondary hair follicles was CSNKIE CSNKIDLBHLHE40NR1D1PER1, CRY1CRY2BHL41CLOCKPER2 PER2 PER3.2CSNKIDOCK-CRY2PER3, CRY2PER3, CRY2PER3, CRY2PER3, CRY2PER3, and CRY2PER3, respectively, and CRY1CRY2BHLHE41CLOCKPER2PER2PER3.2CSNKIDOCKCK-CRY2PER3, CRY2PER3, CRY2PER3, CRY2PER3 and CRY2PER3, respectively. There were significant differences in the expression of NR1D1 gene in the hair follicles of cashmere goats with the growth period and sampling time of hair follicles. The expression of NR1D1 gene in the same growth stage of primary hair follicles showed significant difference. The expression of ARNTL and PER2 genes varied with the secondary stage. The growth period of hair follicles is different, The expression of clock gene in the secondary hair follicles of cashmere goats was significantly different (fluorescent quantitative PCRs) was 1: CLOCK TIM PER1 CRY1.4, and the high amplitude rhythms of PER1 gene, PER1 gene, PER1 gene in the flourishing stage of skin secondary hair follicles were observed in the order of 1: CLOCK PER1 CRY1.4. At 04: 00, 08: 00, 08: 00, 08: 00, 16: 00: 00, respectively, for "reporting the hour"; for the rest period of the skin's secondary hair follicles, The high amplitude rhythmic expression of the four clock genes was carried out at 08: 00: 04: 04: 04: 00 and 16: 00 respectively.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827

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