本文選題：絨山羊　切入點：初級毛囊　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：山羊絨毛生長發(fā)育呈明顯的季節(jié)性變化,它的皮膚作為感知外環(huán)境溫度變化和水分缺失的交界面,能夠使機體內(nèi)環(huán)境與外環(huán)境相協(xié)同發(fā)展。這表明絨山羊皮膚中可能存在著一個時間調(diào)控機制,而這一時間機制受下丘腦視交叉上核(SCN)上生物鐘基因的調(diào)控。因此,生物鐘基因之間相互作用的關(guān)系影響著皮膚毛囊的生長發(fā)育。本研究為進(jìn)一步證實生物鐘基因是否形成一個近乎以24小時為周期的轉(zhuǎn)錄翻譯反饋循環(huán)系統(tǒng)來調(diào)控皮膚毛囊的生長發(fā)育,以13個生物鐘基因CSNKIE、CENK1D、TIM、CLOCK、 PER1、PER2、PER3、TIM、CRY1、CRY2、BHLHE40、BHLHE41、ARNTL、NR1D1為研究對象,采用RNA-Seq技術(shù)初步分析不同生長時期初級毛囊和次級毛囊上生物鐘基因的表達(dá)模式；此外,為深入探究CLCOK、TIM, PER1、CRY1基因在皮膚次級毛囊中的相互作用及其分子機制,文章進(jìn)而應(yīng)用熒光定量PCR技術(shù)對以上4個鐘基因在次級毛囊上的表達(dá)做了定量分析。通過以上研究分析,本文得到如下結(jié)果：1、CSNKIE基因在皮膚毛囊中進(jìn)行高表達(dá),其表達(dá)量高出其它鐘基因幾十倍。生物鐘基因在山羊皮膚初級毛囊上的表達(dá)量依次為：CSNKIE、CSNKID、BHLHE40、ARNTL CRY1 PER1、CLOCK、CRY2、NR1D1 PER2 PER3、BHLHE41;皮膚次級毛囊上的表達(dá)量按從大到小的順序依次為：CSNKIE CSNKID、ARNTL、BHLHE40、NR1D1、PER1、 CRY1、CRY2、BHLHE41、CLOCK、PER2 PER3。2、CSNKID、CLOCK、CRY2、PER3和BHLHE41基因在絨山羊皮膚毛囊上的表達(dá)隨毛囊生長時期和采樣時間的變化存在顯著差異；NR1D1基因在初級毛囊同一生長時期不同采樣時間的表達(dá)差異顯著,ARNTL和PER2基因隨次級毛囊生長時期的不同,其表達(dá)量有顯著差異。3、生物鐘基因在絨山羊皮膚次級毛囊中的表達(dá)量(熒光定量PCR)依次為：CLOCK TIM PER1 CRY1。4、皮膚次級毛囊興盛期,CLOCK基因、TIM基因、PER1基因、CRYl基因的高振幅節(jié)律表達(dá)分別于04：00、08：00、08：00、16：00進(jìn)行“報時”；皮膚次級毛囊休止期,這四個鐘基因的高振幅節(jié)律表達(dá)分別于08：00、04：00、04：00、16：00進(jìn)行“報時”。
[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.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S827

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本文編號：1615819