湖羊不同发育期瘤胃形态学变化及基因表达差异研究

doi:10.11843/j.issn.0366-6964.2025.08.019

摘要/Abstract

摘要：

旨在阐明不同发育阶段湖羊瘤胃组织形态发生与基因表达调控的协同规律，初探瘤胃功能成熟的分子基础。因此，本研究以健康母湖羊及其所产母羔羊为研究对象，按胚胎早期(85±5 d)、胎儿期(125±5 d)、新生期、成熟期(120±5 d)分4组，每组5只。经标准化饲养管理后，电休克处死羔羊并采集瘤胃背囊组织，部分样本经固定染色进行HE观察，分析形态学差异；其余样本冻存用于RNA提取及转录组测序，探究基因表达模式。结果表明，虽然瘤胃壁厚度随着日龄增加会逐渐增厚，但瘤胃乳头发育关键期是新生期至成熟期间。转录组结果也表明，成熟期的瘤胃基因表达模式与其他3个时期有显著差异，差异基因的表达模式可以分为21个典型模块，而成熟期的部分模块主要参与了脂肪酸代谢、免疫屏障构建、矿物质吸收等生物学过程与信号通路。PIP基因在成熟后瘤胃中的表达水平较其他3个发育阶段显著上升，进而调控ErbB2与MAPK等信号通路以完成免疫屏障构建，在成熟期MCT1等溶质载体家族成员表达水平显著上调，促进短链脂肪酸及其代谢物的跨膜运输并维持细胞内pH，完善营养物质转运的分子基础。因此，本研究发现瘤胃成熟的关键过程发生在出生后，基因表达模式的转变支撑了瘤胃形态的成熟，为揭示羔羊瘤胃发育的分子机制提供了一种新的思路。

关键词: 湖羊羔羊, 瘤胃发育, 转录组测序

Abstract:

This study aimed to elucidate the synergistic interplay between rumen tissue morphogenesis and gene expression regulation in Hu sheep across distinct developmental stages, and to preliminarily explore the molecular basis underlying rumen functional maturation. Therefore, this study used healthy female Hu sheep and their female lambs as research subjects, which were divided into 4 groups according to the embryonic period (85±5 d), fetal period (125±5 d), neonatal period, and mature period (120±5 d), with 5 individuals in each group. After standardized feeding management, the lambs were euthanized by electroshock, and rumen dorsal sac tissues were collected. Part of the samples were fixed and stained for HE observation to analyze morphological differences; the remaining samples were frozen for RNA extraction and transcriptome sequencing to explore gene expression patterns. The results showed that although the thickness of the rumen wall gradually increased with age, the critical period for rumen papilla development occurred from the neonatal period to the mature stage. Transcriptome results also indicated that the rumen gene expression pattern at the mature stage was significantly different from that of the other 3 stages. The expression patterns of differentially expressed genes could be divided into 21 typical modules, among which some modules in the mature stage were mainly involved in biological processes and signaling pathways such as fatty acid metabolism, immune barrier construction, and mineral absorption. The expression level of the PIP gene in the rumen at maturity was significantly higher than that in the other 3 developmental stages, thereby regulating signaling pathways such as ErbB2 and MAPK to complete immune barrier construction. At the mature stage, the expression levels of solute carrier family members such as MCT1 were significantly upregulated, promoting the transmembrane transport of short-chain fatty acids and their metabolites, maintaining intracellular pH, and improving the molecular basis of nutrient transport. Therefore, the present study found that the key process of rumen maturation occurs after birth, and the transformation of gene expression pattern supports the maturation of rumen morphology, which provides a new way of thinking to reveal the molecular mechanism of rumen development in lambs.

Key words: Hu lamb, rumen development, transcriptome sequencing

中图分类号:

S826.2

范婧, 李伟, 朱妍, 勿都巴拉, 史佳慧, 胡斯乐, 吴江鸿. 湖羊不同发育期瘤胃形态学变化及基因表达差异研究[J]. 畜牧兽医学报, 2025, 56(8): 3773-3786.

FAN Jing, LI Wei, ZHU Yan, Wudubala , SHI Jiahui, Husile , WU Jianghong. Study on Rumen Morphological Changes and Gene Expression Differences in Hu Sheep at Different Developmental Stages[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3773-3786.

图/表 11

表 1

表 2

图 1

表 3

表 4

图 2

图 3

表 5

KEGG、GO注释结果"

分类 Category	编号 ID	条目 Term	模块 ME
脂肪酸代谢相关功能注释Functional annotations related to fatty acid metabolism
BP	GO: 0033539	酰基辅酶A脱氢酶介导的脂肪酸β -氧化	7
BP	GO: 0006635	脂肪酸β -氧化	7
BP	GO: 0019367	脂肪酸延长	1
CC	GO: 0005739	线粒体	1、7、20
CC	GO: 0005783	内质网	1、4、20
CC	GO: 0031410	胞质囊泡	4、7
MF	GO: 0009922	脂肪酸延长酶活性	1
MF	GO: 0070403	NAD⁺结合	1、4
MF	GO: 0004497	单加氧酶活性	16
KEGG	oas01212	脂肪酸代谢	1、7
KEGG	oas00071	脂肪酸降解	1、7
KEGG	oas00062	脂肪酸延长	1、7
KEGG	oas01240	辅因子生物合成	1、4、7、16
KEGG	oas00640	丙酸代谢	7
免疫屏障相关功能注释Functional annotations related to immune barrier
BP	GO: 0071222	细胞对脂多糖的应答	4
BP	GO: 0030855	上皮细胞分化	4
BP	GO: 0050829	对革兰氏阴性菌的防御反应	4
CC	GO: 0031012	细胞外基质	1、4
CC	GO: 0005765	溶酶体膜	4、16
CC	GO: 0008305	整合素复合体	15
MF	GO: 0005149	白细胞介素-1受体结合	4
MF	GO: 0003796	溶菌酶活性	4
MF	GO: 0050839	细胞黏附分子结合	1
KEGG	oas00980	细胞色素P450对外源物的代谢	1
KEGG	oas04520	黏着连接	1
KEGG	oas04530	紧密连接	4
KEGG	oas00480	谷胱甘肽代谢	1、15
KEGG	oas05150	金黄色葡萄球菌感染	4
物质转运相关功能注释Functional annotations related to substance transport
BP	GO: 0006730	一碳代谢	1、4、15
BP	GO: 0055085	跨膜运输	4
BP	GO: 0006814	钠离子转运	4
CC	GO: 0005794	高尔基体	1
CC	GO: 0005922	连接蛋白复合体	1
CC	GO: 0016324	顶质膜	4
MF	GO: 1903763	参与电耦联细胞通讯的缝隙连接通道活性	1
MF	GO: 0005246	钙通道调节活性	15
MF	GO: 0046872	金属离子结合	1、7、16
KEGG	oas04976	胆汁分泌	1
KEGG	oas04978	矿物质吸收	4
KEGG	oas04530	紧密连接	4
KEGG	oas04520	黏着连接	1
KEGG	oas01200	碳代谢	4

表 5

图 4

图 5

图 6

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项目 Item	妊娠母羊 Pregnant ewe	断奶羔羊 Weaned lamb
原料组成 Ingredient
玉米 Maize	55.50	55
豆粕 Bean meal	20	26
DDGS	8	8
预混料 Premix	5	5
小苏打 NaHCO₃	1	1
玉米胚芽粕 Corn germ oil meal	10	-
食盐 Salt	0.5	-
棉籽粕 Cotton seed meal	-	5
合计 Total	100	100

基因 Gene	登录号 Accession number	引物序列(5′→3′) Primer sequence	产物大小/bp Size	退火温度/℃ Annealing temperature
PRDM1	XM_004011238.5	F: CACCACTTCATTGATGGCTTTA	146	60
		R: TTGGCAGGGATGGGCTTA
PRDM10	XM_042237978.1	F: TTCAAGTGCCGCCTCTGC	134	60
		R: TCACGGTTGGGCTTTATG
PRDM11	XM_042233404.2	F: GATGGTGACGGTGGTGAA	146	60
		R: TTTTGGGCATGATGAGGT
HDAC2	XM_004011189.4	F: TTATTACTACGGACAGGG	125	58
		R: CTTCAGCAGTGGCTTTAT
LOC101122351	XM_027968888.2	F: CCCTGCTTCTGATTCTTTG	110	60
		R: CTCTTCACTCCCGTCTATTT
SH3RF2	XM_004008919.5	F: TCAAGAAACAGCAGCGAGAA	170	63
		R: CGGTGGGCGAACAAGATG
IGF1	NM_001009774.3	F: AATCAGCAGTCTTCCAACCCAA	120	62
		R: AAGGCGAGCAAGCACAGG
SLC16A1	XM_046946303.1	F: ACCAGTTTTAGGTCGTCTCA	207	60
		R: GGCTTCTCAGCAACATCTACA
β-Actin	NM_001009784.3	F: TGGACTTCGAGCAGGAGATG	139	60
		R: AGGAAGGAAGGCTGGAAGAG

项目 Item	胚胎期 Embryonic period	胎儿期 Fetal period	新生期 Newborn period	成熟期 Mature period
乳头长度/mm Nipple length	0.09±0.001^Cc	0.13±0.003^Cc	0.32±0.008^Bb	2.46±0.027^Aa
乳头宽度/mm Nipple width	0.03±0.003^Cd	0.06±0.003^Bc	0.08±0.004^Bb	0.71±0.007^Aa
乳头密度/(个·cm^-2) Nipple density	471.33±4.097^Aa	394.67±3.180^Bb	292.67±2.428^Cc	42.67±2.603^Dd
乳头表面积/(mm²·cm^-2) Nipple surface area	2.37±0.142^Bb	6.47±0.183^Bb	16.70±0.177^Bb	148.48±9.84^Aa
总厚度/μm Total thickness	30.86±0.820^Dd	50.78±1.066^Cc	70.55±1.676^Bb	247.60±2.52^Aa
角质层/μm Stratum corneum	4.52±0.366^Bc	8.19±0.189^Bb	9.11±0.166^Bb	41.82±1.477^Aa
颗粒层/μm Granular layer	8.47±0.303^Cc	13.83±1.138^Bb	16.28±0.263^Bb	68.60±1.168^Aa
棘层和颗粒层/μm Spinous layer and granular layer	17.94±0.511^Dd	28.76±0.382^Cc	45.16±1.517^Bb	137.18±1.07^Aa

样本名 Sample	高质量序列/Mb Clean read	高质量碱基数/Gb Clean base	Q20/%	Q30/%	GC/%
ER1	21.67	6.33	97.51	93.13	50.78
ER2	22.96	6.68	97.35	92.65	50.56
ER3	22.07	6.44	97.60	93.30	51.52
ER4	21.75	6.36	96.75	90.97	50.05
ER5	20.99	6.14	96.62	90.56	49.81
LR1	21.01	6.15	96.61	90.58	49.70
LR2	21.28	6.22	96.69	90.70	50.74
LR3	23.22	6.80	96.65	90.60	49.86
LR4	22.98	6.72	96.87	91.38	50.46
LR5	21.19	6.21	96.71	90.85	50.32
FR1	22.66	7.12	97.25	92.70	48.20
FR2	25.55	8.00	97.09	92.28	46.51
FR3	20.56	6.46	97.00	92.02	49.01
FR4	21.13	6.64	97.65	93.40	49.05
FR5	26.50	8.31	97.41	93.13	49.33
AR1	21.34	6.24	96.79	90.96	48.22
AR2	21.66	6.34	96.95	91.39	49.23
AR3	20.18	5.90	96.80	90.96	49.08
AR4	20.40	5.97	96.74	90.87	49.94
AR5	20.92	6.12	96.45	90.10	47.59

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