Urine differential metabolites and functional prediction of male Bactrian camel (Camelus ferus) in estrus and anestrus

doi:10.16829/j.slxb.150951

Abstract

Abstract:

The aim of this study was to explore the differences of urine metabolism in male Bactrian camels (Camelus ferus) during estrus, in order to provide theoretical basis for the reproductive regulation mechanism of Bactrian camels. Urine samples from breeding season (BS) and non-breeding season (NBS) male Bactrian camels were collected using Gas/Liquid chromatography-mass spectrometry (GC/LC-MS) techniques. Screening for Differentially expressed metabolites (DEMs) in urine; Based on the Human Metabolome Database (HMDB) and the Kyoto Encyclopedia of Genes and Genomes, KEGG database to identify key functional substances in urine that influence seasonal estrus in Bactrian camels and their pathways. The results showed that 468 metabolites were identified by GC-MS and 6 193 by LC-MS. Compared with NBS, there were 205 DEMs and 601 DEMs identified by GC/LC-MS in BS group, respectively (P < 0.05), mainly of which were benzene and its substituted derivatives, fatty acyl, indole and its derivatives, and organic nitrogen compounds. Twelve identical pathways were obtained by GC/LC-MS KEGG enrichment analysis, and five common DEMs were obtained by GC/LC-MS Wien: L-histidine, 5′ -methyl-adenosine, 3-methylhistidine, L-methionine and triacetin. The results showed that L-histidine and L-methionine could affect spermatogenesis and reproductive health through amino acid tRNA biosynthesis and histidine metabolism pathway, and further affect the estrus of male Bactrian camels. In addition, based on the roles of L-histidine and L-methionine in female animals, it is speculated that these DEMs may act as pheromones to affect the synthesis of sex hormones in female camels, and thus induce estrus in female camels. KEGG enrichment analysis showed that male Bactrian camel estrus was also related to sperm motility and reproductive health regulated by TCA cycle, arginine biosynthesis pathway, GABA signaling and pyruvate metabolism. The above results provide an important reference for further analysis of the reproductive regulation mechanism of Bactrian camels.

Key words: Bactrian camel (Camelus ferus), Seasonal estrus, Urine, GC/LC-MS, Differentially expressed metabolites, Functional predictions, Pheromone

摘要：

为探究发情期雄性双峰驼 (Camelus ferus) 尿液的代谢差异，以期为双峰驼繁殖调控机制提供理论依据。本研究采集发情期 (Breeding season, BS) (n = 6) 和非发情期 (Non‑breeding season, NBS) (n = 6) 雄性双峰驼尿液，利用气相/液相色谱-质谱 (Gas/Liquid chromatography‑mass spectrometry, GC/LC‑MS) 技术，筛选尿液中差异表达代谢物 (Differentially expressed metabolites, DEMs)；基于人类代谢组数据库 (Human Metabolome Database, HMDB) 和京都基因与基因组百科全书 (Kyoto Encyclopedia of Genes and Genomes, KEGG) 数据库鉴定尿液中影响双峰驼季节性发情的关键功效物质及其通路。结果显示，GC‑MS鉴定代谢物468个，LC‑MS鉴定代谢物6 193个。以NBS为对照，BS组GC/LC‑MS分别鉴定到205个和601个DEMs (P < 0.05)，主要为苯及其取代衍生物、脂肪酰、吲哚及其衍生物和有机氮化合物等。GC/LC‑MS KEGG富集分析鉴定获得12条相同通路，GC/LC‑MS维恩鉴定获得L-组氨酸、5'-甲硫基腺苷、3-甲基组氨酸、L-蛋氨酸和三醋精5种共有DEMs。结果表明，L-组氨酸和L-蛋氨酸可通过氨基酸-tRNA的生物合成和组氨酸代谢通路，影响精子发生和生殖健康，从而进一步影响雄性双峰驼发情。此外，基于L-组氨酸和L-蛋氨酸在雌性动物中的作用，推测这两种DEMs可能作为信息素影响雌性双峰驼的性激素合成，进而诱导母驼发情。KEGG富集分析表明，雄性双峰驼发情还与TCA循环、精氨酸的生物合成途径、GABA能信号传导及丙酮酸代谢调控的精子活力和生殖健康有关。上述结果为深入解析双峰驼繁殖调控机制研究提供了重要参考。

关键词: 双峰驼, 季节性发情, 尿液, 气相/液相色谱-质谱, 差异代谢物, 功能预测, 信息素

CLC Number:

Q592.2

Bao YUAN, Bohao ZHANG, Jianfu LI, Wenli CHEN, Yong ZHANG, Xingxu ZHAO, Quanwei ZHANG. Urine differential metabolites and functional prediction of male Bactrian camel (Camelus ferus) in estrus and anestrus[J]. ACTA THERIOLOGICA SINICA, 2025, 45(3): 399-408.

袁宝, 张博皓, 李建富, 陈文丽, 张勇, 赵兴绪, 张全伟. 发情和非发情期雄性双峰驼尿液差异代谢物及其功能预测[J]. 兽类学报, 2025, 45(3): 399-408.

Figures/Tables 5

Fig. 1 Urine analysis of GC-MS data from Camelus ferus. A: TIC overlay diagram of QC sample; B: Boxplot of metabolite strength; C: PCA score plots for all samples; D: OPLS-DA model replacement test plot. R2: Coefficient of determination; Q2: Predictability. QC: Quality control; BS: Breeding season; NBS: Non-breeding season

Fig. 2 Urine analysis of LC-MS data from Camelus ferus. A: PCA score plots for all samples; B: Boxplot of metabolite strength; C: Splot diagram; D: OPLS-DA model replacement test plot; R2 Coefficient of determination; Q2: Predictability. QC: Quality control; BS: Breeding season; NBS: Non-breeding season

Fig. 3 DEMs identified of urine from Camelus ferus by GC/LC-MS and classification. A, B, C: DEMs in GC-MS; D, E, F: DEMs in LC-MS. BS: Breeding season; NBS: Non-breeding season

Fig. 4 Urine analysis of GC/LC-MS common DEMs from Camelus ferus. A: the common DEMs in GC/LC-MS; B: the common DEMs in GC-MS; C: the common DEMs in LC-MS.BS: Breeding season; NBS: Non-breeding season

Fig. 5 Metabolic pathway enrichment analysis of urine DEMs from Camelus ferus. A: GC-MS top20 pathways; B: LC-MS top20 pathways; C: LC-MS top20 pathways; D: the common DEMs pathway

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