乏情和发情初产母猪下丘脑-垂体-卵巢轴中lincRNAs表达谱比较分析

doi:10.16288/j.yczz.19-347

遗传 ›› 2020, Vol. 42 ›› Issue (4): 388-402.doi: 10.16288/j.yczz.19-347

乏情和发情初产母猪下丘脑-垂体-卵巢轴中lincRNAs表达谱比较分析

任巧玲¹, 张家庆¹, 陆东锋², 王璟¹, 陈俊峰¹, 马强¹, 白献晓¹, 郭红霞¹, 高彬文¹, 邢宝松¹()

^1. 河南省农业科学院畜牧兽医研究所,河南省畜禽繁育与营养调控重点实验室,郑州 450002
^2. 河南花花牛实业总公司,郑州 450000

收稿日期:2019-11-14 修回日期:2020-02-29 出版日期:2020-04-20 发布日期:2020-03-26
通讯作者: 邢宝松 E-mail:baosong@126.com
作者简介:任巧玲,硕士,副研究员,研究方向：猪的育种与营养。E-mail: renql76@163.com;
基金资助:
河南省农业科学院科技创新创意项目编号(2020CX06);河南省重点研发专项编号(182102110063);河南省财政预算科研专项项目编号(2019CY015);河南省农业科学院自主创新项目资助编号(2019ZC42)

Comparison and analysis of lincRNAs expression profile in the hypothalamic-pituitary-ovarian axis of anestrous and estrous primiparous sows

Qiaoling Ren¹, Jiaqing Zhang¹, Dongfeng Lu², Jing Wang¹, Junfeng Chen¹, Qiang Ma¹, Xianxiao Bai¹, Hongxiao Guo¹, Binwen Gao¹, Baosong Xing¹()

^1. Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation , Institute of Animal Husbandry and Veterinary Science, Henan Academy of Agricultural Sciences, Zhengzhou 450002,China
^2. Henan Huahuaniu Industrial Corporation, Zhengzhou 450000, China

Received:2019-11-14 Revised:2020-02-29 Online:2020-04-20 Published:2020-03-26
Contact: Xing Baosong E-mail:baosong@126.com
Supported by:
Supported by Science and Technology Innovation Program No(2020CX06);Key Special Rresearch and Development Program of Henan Province No(182102110063);Henan Province Financial Budget for Scientific Research Program No(2019CY015);the Program for Independent Innovative Research in Henan Academy of Agricultural Sciences No(2019ZC42)

摘要/Abstract

摘要：

初产母猪断奶后能否正常发情对养猪生产影响重大,也是初产母猪被淘汰的主要原因。本研究以乏情和发情初产母猪为研究对象,首次利用RNA-seq技术对其下丘脑-垂体-卵巢轴中的基因间长链非编码RNAs(long intergenic noncoding RNAs, lincRNAs)进行筛选比较,得到lincRNAs的表达图谱,并对其特征和功能进行了初步分析。结果显示,在乏情和发情初产母猪下丘脑-垂体-卵巢轴中鉴定得到3519个lincRNAs,以发情组为对照共有17个lincRNAs存在差异表达,其中12个表达上调,5个表达下调(FC≥2, P<0.05)。选择4个差异表达的lincRNAs经qRT-PCR验证,其表达水平与测序结果基本一致。对这17个差异表达的lincRNAs进行GO分析、KEGG通路分析及lincRNA-mRNA共表达网络分析,发现这些lincRNAs主要与猪卵母细胞减数分裂成熟、卵巢细胞分化及颗粒细胞凋亡等生殖活动相关。本研究结果丰富了猪lincRNAs数据资源,为进一步深入研究初产母猪的生殖机能提供了理论依据。

关键词: lincRNAs, 乏情, 发情, 初产母猪, 下丘脑-垂体-卵巢轴

Abstract:

The normal estrus in weaned primiparous sows has a great impact on pig production and abnormal estrus is the main reason for the elimination of primiparous sows. In this study, we studied the long intergenic noncoding RNAs (lincRNAs) in the hypothalamic-pituitary-ovarian axis of anestrous and estrous primiparous sows. These long intergenic noncoding RNAs (lincRNAs) were screened and compared through RNA-seq analysis. The expression profiles of lincRNAs were obtained and their characteristics and functions were preliminarily analyzed. There are 3519 novel lincRNAs identified in the hypothalamic-pituitary-ovarian axis of anestrous and estrous primiparous sows. Compared with estrous primiparous sows, 17 differentially expressed lincRNAs were indentified, including 12 up-regulated lincRNAs and 5 down-regulated lincRNAs (FC≥2, P<0.05). The four lincRNA transcripts obtained through selection were verified by qRT-PCR, which are consistent with the RNA-seq results. The GO, KEGG pathway, and lincRNA-mRNA co-expression network analysis of these 17 lincRNAs revealed that these lincRNAs were mainly involved in reproductive activities, such as oocyte meiosis mature, ovarian cells differentiation and granulosa cells apoptosis. The results enriched the data resources of pig lincRNAs and provided useful information for further research about the reproductive performance of primiparous sows.

Key words: lincRNAs, anestrus, estrus, primiparous sows, hypothalamic-pituitary-ovarian axis

任巧玲, 张家庆, 陆东锋, 王璟, 陈俊峰, 马强, 白献晓, 郭红霞, 高彬文, 邢宝松. 乏情和发情初产母猪下丘脑-垂体-卵巢轴中lincRNAs表达谱比较分析[J]. 遗传, 2020, 42(4): 388-402.

Qiaoling Ren, Jiaqing Zhang, Dongfeng Lu, Jing Wang, Junfeng Chen, Qiang Ma, Xianxiao Bai, Hongxiao Guo, Binwen Gao, Baosong Xing. Comparison and analysis of lincRNAs expression profile in the hypothalamic-pituitary-ovarian axis of anestrous and estrous primiparous sows[J]. Hereditas(Beijing), 2020, 42(4): 388-402.

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样品	Raw reads	Clean reads	Q30 (%)	Mapped reads	Mapping ratio (%)
乏情下丘脑	53,306,278	52,516,724	97.14	43,261,322	82.38
发情下丘脑	53,268,878	52,432,800	97.05	41,257,875	78.69
乏情垂体	53,334,742	52,476,820	96.96	43,582,111	83.05
发情垂体	5,331,090	52,573,730	97.14	42,844,655	81.49
乏情卵巢	53,216,626	52,195,962	96.70	41,861,577	80.20
发情卵巢	53,222,368	52,209,252	96.70	41,381,612	79.26

lincRNA	引物序列(5′→3′)	复性温度(℃)	扩增长度(bp)
TCONS_00049640	F：TTCTCATAGTCCCCACACAGCC	60	177
TCONS_00049640	R：CTTCTTCTTTCTCTTCCTTCCCC	60	177
TCONS_00001098	F：TGAGGAAGAGCAGATAGCCAGTC	60	224
TCONS_00001098	R：AGGTATTCAGGGAGGGCATCTA	60	224
TCONS_ 00005732	F：TCCTTTCAGATTGGCTGGTGTT	60	230
TCONS_ 00005732	R：CCACAGTCCAGGTCATCAAGC	60	230
TCONS_00012288	F：GCCCAGTGGATTTCATTTGC	60	185
TCONS_00012288	R：TGATACCATTCCTTCCTGTTCTCC	60	185
GAPDH	F：GTGAAGGTCGGAGTGAACGGA	60	252
GAPDH	R：CCATTTGATGTTGGCGGGAT	60	252

转录本ID	基因ID	平均表达量		P值	\|log₂FC\|	调控	位置
转录本ID	基因ID	乏情组	发情组	P值	\|log₂FC\|	调控	位置
TCONS_00035245	XLOC_018088	51.963	0.001	0.003	15.665	上调	Chr.4(+):139486869~139488925
TCONS_00006245	XLOC_003209	38.671	0.001	0.012	15.239	上调	Chr.10(+):76375911~76390174
TCONS_00038863	XLOC_019956	31.061	0.001	0.014	14.923	上调	Chr.5(-):46693872~46740946
TCONS_00039679	XLOC_020371	29.799	0.001	0.017	14.863	上调	Chr.6(+):4250343~:4255208
TCONS_00049640	XLOC_025373	68.476	3.245	0.024	4.399	上调	Chr.8(-):31318960~31504408
TCONS_00041695	XLOC_021350	36.005	0.611	0.026	5.881	上调	Chr.6(+):91775209~:91809105
TCONS_00041696	XLOC_021350	36.005	0.611	0.026	5.881	上调	Chr.6(+):91802418~91809105
TCONS_00040987	XLOC_020986	24.830	0.001	0.027	14.600	上调	Chr.6(+):57220929~57255522
TCONS_00031805	XLOC_016327	63.453	3.241	0.029	4.291	上调	Chr.3(+):140652352~140654672
TCONS_00027978	XLOC_014327	60.319	3.346	0.034	4.172	上调	Chr.2(-):8241783~8244938
TCONS_00001098	XLOC_000568	103.316	9.467	0.042	3.448	上调	Chr.1(+):137503074~137507662
TCONS_00029995	XLOC_015378	26.051	0.306	0.045	6.414	上调	Chr.3(+):4643111~:4645116
TCONS_00032971	XLOC_016898	0.001	75.785	0.001	16.210	下调	Chr.3(-):60770406~:60792096
TCONS_00025231	XLOC_012981	0.317	30.704	0.036	6.600	下调	Chr.18(-):57407991~57423170
TCONS_00009672	XLOC_005062	0.001	26.692	0.040	14.704	下调	Chr.12(-):952204~:953646
TCONS_00005732	XLOC_002919	102.234	672.631	0.048	2.718	下调	Chr.10(+):22597153~22603044
TCONS_00012288	XLOC_006336	13.226	78.923	0.049	2.577	下调	Chr.13(+):128701400~128715093

转录本ID	基因名称	平均表达量		P值	\|log₂FC\|	调控
转录本ID	基因名称	乏情组	发情组	P值	\|log₂FC\|	调控
ENSSSCT00000031961		125.320	3.532	0.001	5.149	上调
ENSSSCT00000036099	TRAPPC9-002	142.211	1.300	0.012	6.744	上调
ENSSSCT00000022861	UQCC3-201	655.643	62.058	0.006	3.401	上调
ENSSSCT00000011345		404.656	37.820	0.007	3.420	上调
ENSSSCT00000015759		202.943	14.773	0.008	3.780	上调
ENSSSCT00000001351	PPP1R11-001	820.812	96.141	0.011	3.094	上调
ENSSSCT00000028568	C14H10orf116-201	1129.122	18.993	0.031	5.894	上调
ENSSSCT00000025288		202.590	5.782	0.038	5.131	上调
ENSSSCT00000036373	TSPAN6-002	602.708	21.985	0.042	4.777	上调
ENSSSCT00000001615	SLA-DRB1-201	229.961	4.358	0.046	5.722	上调
ENSSSCT00000016351	MMP13-201	3706.129	5.799	0.049	9.320	上调
ENSSSCT00000012778		18.317	803.462	7.06E-05	5.455	下调
ENSSSCT00000022502		7.887	220.445	0.001	4.805	下调
ENSSSCT00000000207	TUBA1C-201	552.531	7980.832	0.002	3.852	下调
ENSSSCT00000032751	TXLNG-002	16.643	238.873	0.003	3.843	下调
ENSSSCT00000033602	PGRMC1-003	22.350	184.633	0.020	3.046	下调
ENSSSCT00000029673		75.697	434.230	0.035	2.520	下调

乏情和发情初产母猪下丘脑-垂体-卵巢轴中lincRNAs表达谱比较分析

Comparison and analysis of lincRNAs expression profile in the hypothalamic-pituitary-ovarian axis of anestrous and estrous primiparous sows

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