光照强度对樱桃谷肉鸭c-fos、生物钟基因表达 及褪黑激素的影响

doi:10.3864/j.issn.0578-1752.2020.04.016

摘要/Abstract

摘要：

【目的】研究持续黑暗下给予樱桃谷肉鸭不同强度的瞬时光照,对下丘脑、间脑、垂体和中脑中c-fos基因及相关生物钟基因的表达及血浆和肝脏中褪黑激素含量的影响。【方法】选用144只健康且体重接近的1日龄樱桃谷肉鸭饲养至21日龄,22日龄时随机分为2组,每组6个重复,持续黑暗7 d,期间自由采食和饮水,分别采用10 Lx和80 Lx的LED白光刺激1.5 h,暗适应1.5 h后立即从每个重复选取1只接近平均体重的肉鸭,检测下丘脑、间脑、垂体和中脑c-fos和生物钟基因的表达量,检测血浆和肝脏褪黑激素含量。【结果】10 Lx组的下丘脑和中脑c-fos表达量显著高于80 Lx光照组（P＜0.05）,间脑中c-fos的表达量显著低于80 Lx光照组（P＜0.05）,两组垂体组织中c-fos表达无显著影响（P＞0.05）。与80 Lx组相比,10 Lx组下丘脑生物钟基因Bmal1、Bmal2、Clock和Per2表达量均显著提高（P＜0.05）,Cry1表达量也有提高的趋势。10 Lx组间脑Bmal1的表达量显著提高（P＜0.05）,Bmal2、Clock、Per2和Cry1的表达量无显著差异（P＞0.05）。10 Lx组中脑Clock和Per2的表达量显著高于80 Lx组（P＜0.05）,两组Bmal1、Bmal2和Cry1表达无显著影响（P＞0.05）。不同强度的瞬时光照对垂体中生物钟基因的表达无显著影响（P＞0.05）。与80 Lx组相比,10 Lx组肉鸭血浆中褪黑激素含量显著降低（P＜0.05）,肝脏中褪黑激素含量显著升高（P＜0.05）。【结论】持续黑暗下,短暂高光强刺激可抑制下丘脑和中脑c-fos的表达,而提高间脑c-fos基因的表达,同时减弱了肉鸭下丘脑和间脑生物钟基因表达的振幅。

关键词: 樱桃谷肉鸭, 持续黑暗, 光照强度, c-fos, 生物钟

Abstract:

【Objective】 The objective of this study was to investigate the effects of different intensities of light stimulation on c-fos and related clock genes in hypothalamus, diencephalon, pituitary and mesencephalon, as well as melatonin levels in plasma and liver of cherry valley meat duck under continuous darkness. 【Method】 A total of one hundred and forty four 1-day-old cherry valley meat ducks were raised to the age of 21 days. 22-day-old cherry valley meat ducks were randomly divided into 2 groups. Each group had 6 replicates, with 12 ducks per replicate, and kept dark for 7 days, the ducks were given ad libitum access to water and feed. After 7 days lasting darkness, 28-day-old ducks received a LED white light stimulation of different intensities for 1.5 h and then a dark adaptation for 1.5 h. One duck from each replicate was slaughted for sample collection, the expression levels of c-fos and circadian clock genes in the hypothalamus, diencephalon, pituitary and mesencephalon were measured, and plasma and liver melatonin levels were measured. 【Result】 The expression level of c-fos in hypothalamus and mesencephalon of the 10 Lx group was significantly higher than that in the 80 Lx group (P＜0.05). The level of c-fos in the diencephalon was significantly lower than that in the 80 Lx group (P＜0.05), and there was no significant difference of c-fos expression in pituitary in the two groups(P＞0.05). Compared with the 80 Lx group, the expression of clock gene clock, Bmal1, Bmal2 and Per2 in the 10 Lx group in the hypothalamus was significantly increased (P＜0.05), and the expression of Cry1 was little increased in the 10 Lx group (P=0.06) .The expression level of Bmal1 in the diencephalon was significantly higher in the 10 Lx group (P＜0.05), while Bmal2, Clock, Per2 and Cry1 showed no significant difference (P＞0.05) in two groups. The expression of Clock and Per2 in the mesencephalon was significantly higher in the 10 Lx group than that in the 80 Lx group (P＜0.05). The expression of Bmal1, Bmal2 and Cry1 in the two groups had no significant difference (P＞0.05). The expression of clock genes in pituitary was not significant affect by light intensity (P＞0.05). Compared with the 80 Lx, the plasma level of melatonin in 10 Lx group was significantly lower (P＜0.05), while the melatonin levels in the liver were significantly higher. 【Conclusion】 In the continuous darkness, transient high intensity light stimulation could inhibit the expression of c-fos in the hypothalamus and mesencephalon, and increase the expression of c-fos in the diencephalon, while the stimulation could attenuate the amplitude of gene expression in the hypothalamus and diencephalon of the meat duck.

Key words: cherry valley meat duck, continuous darkness, light intensity, c-fos, biological clock

崔家杰,谢强,翟双双,龚涛,朱勇文,杨琳,王文策. 光照强度对樱桃谷肉鸭c-fos、生物钟基因表达及褪黑激素的影响[J]. 中国农业科学, 2020, 53(4): 848-856.

JiaJie CUI,Qiang XIE,ShuangShuang ZHAI,Tao GONG,YongWen ZHU,Lin YANG,WenCe WANG. Effects of Light Intensity on c-fos, Biological Clock Gene Expression and Melatonin in Cherry Valley Meat Ducks[J]. Scientia Agricultura Sinica, 2020, 53(4): 848-856.

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基因 Gene	序列号 Accession No.	引物序列（5'-3'） Primer sequence (5′-3′)	大小 Product (bp)
c-fos	NM_205508.1	F: CAGCCCACCCTCATCTCCTC	144
c-fos	NM_205508.1	R: CCTTCGTCCGATGCTCTGC	144
Clock	XM_005012718.3	F: GCACTCCTCCAAGGCAAAG	169
Clock	XM_005012718.3	R: AACACTGGCTGGGACACG	169
Cry1	XM_005011518.2	F: GGCTGTTTGTCTTGTCGT	145
Cry1	XM_005011518.2	R: ACCGTGGATTGTTAGTCG	145
Bmal1	XM_013106919.2	F: TGTTTCCACCAGCGTTCT	133
Bmal1	XM_013106919.2	R: AATGCCAGGGACAGTAGG	133
Bmal2	NM-204133.1	F: CGGCGTTCCTTCTTCTGTC	165
Bmal2	NM-204133.1	R: TTCCTCTTCCACTCCCACC	165
Per2	XM_005028592.3	F: AAGCGTTGTATCACTCAC	106
Per2	XM_005028592.3	R: ACTCGGACCATCTTCTAT	106
β-actin	NM_001310421.1	F: TACGCCAACACGGTGCTG	215
β-actin	NM_001310421.1	R: GATTCATCATACTCCTGCTTG	215

	10 Lx	80 Lx	标准误 Sem	P值 P value
下丘脑Hypothalamus	1.15A	0.87B	0.05	＜0.01
间脑Diencephalon	1.43A	0.76B	0.08	＜0.01
垂体Pituitary	1.20	1.04	0.19	0.60
中脑Mesencephalon	1.26	1.20	0.20	0.87

	10 Lx	80 Lx	标准误 Sem	P值 P value
下丘脑Hypothalamus	1.03A	0.51B	0.09	＜0.01
间脑Diencephalon	1.03	0.72	0.12	0.08
垂体Pituitary	1.05	0.98	0.12	0.74
中脑Mesencephalon	0.89	0.76	0.12	0.45

	10 Lx	80 Lx	标准误 Sem	P值 P value
下丘脑Hypothalamus	0.96a	0.44b	0.13	0.02
间脑Diencephalon	1.09	0.65	0.35	0.19
垂体Pituitary	0.78	0.54	0.10	0.14
中脑Mesencephalon	0.81a	0.50b	0.07	0.02

	10 Lx	80 Lx	标准误 Sem	P值 P value
下丘脑Hypothalamus	0.93	0.48	0.14	0.06
间脑Diencephalon	0.90	0.62	0.10	0.09
垂体Pituitary	1.00	0.76	0.12	0.23
中脑Mesencephalon	1.01	0.86	0.08	0.18