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1.
J Anim Sci ; 1012023 Jan 03.
Artículo en Inglés | MEDLINE | ID: mdl-36331073

RESUMEN

Reproductive seasonality is a limiting factor in sheep production. Sexual behavior is a key element in reproductive efficiency, and this function is regulated by the hypothalamus-pituitary-gonadal (HPG) axis. To understand the mechanisms of sexual behavior, transcriptomic sequencing technology was used to identify differentially expressed genes (DEGs) in the hypothalamus (HT), pars tuberalis (PT) and pineal gland (PG) in Rasa Aragonesa rams with different sexual behavior. Bioinformatics analysis of the 16,401 identified genes by RNA-Seq revealed 103 and 12 DEGs in the HT and the PG, respectively, at a false discovery rate (FDR) of 5% with an absolute value of expression ≥ 1 (log2FC). However, no DEGs were found in the PT. Functional annotation and pathway enrichment analysis showed that DEGs of HT were enriched mainly in neuroactive ligand-receptor interactions and signaling pathways, including notable candidate genes such as MTNR1A, CHRNA2, FSHB, LHB, GNRHR, AVP, PRL, PDYN, CGA, GABRD, and TSHB, which play a crucial role in sexual behavior. The GnRH and cAMP signaling pathways were also highlighted. In addition, gene set enrichment analysis (GSEA) identified potential pathways, dominated mainly by biological process category, that could be responsible for the differences in sexual behavior observed in rams. The intracellular protein transport and pattern specification process were enriched within the PT and the transcription factor binding and protein ubiquitination pathways for the PG. Thus, these pathways together may play an important role in the regulation of the sexual behavior in Rasa Aragonesa rams through the hypothalamic-pituitary-gonadal axis. The validation of 5 DEGs using reverse transcription quantitative polymerase chain reaction (RT-qPCR) showed expression patterns like the found with RNA-Seq. Overall, these results contribute to understanding the genomic basis of sexual behavior in rams. Our study demonstrates that multiple networks and pathways orchestrate sexual behavior in sheep.


Male sexual behavior is a key factor in reproduction, especially in seasonal breeders such as sheep. The identification of differentially expressed genes (DEGs) in brain regions involved in male reproduction and sexual behavior between rams with different sexual activity by RNA high-throughput sequencing can provide useful information to the sheep meat industry. This work aimed to determine the possible molecular mechanisms underlying the sexual behavior of Rasa Aragonesa rams by investigating transcriptional changes in the hypothalamus (HT), pars tuberalis (PT) and pineal gland (PG) between active (A) and nonactive (NA) rams. Comparative analysis revealed 103 and 12 DEGs between the A vs. NA comparison in the HT and the PG, respectively, but no DEGs were found in the PT. Gene ontology (GO) enrichment analysis of DEGs in HT samples revealed significant pathways, associated mainly with neuroactive ligand-receptor interactions, and the GnRH and cAMP signaling pathways. Furthermore, gene set enrichment analysis (GSEA) detected many overrepresented pathways related to sexual behavior via an interaction network within the hypothalamic-pituitary-gonadal axis. These data will be helpful for further investigations to look for mutations or functional single nucleotide polymorphisms (SNPs) that may be used for genetic assisted selection to improve sexual behavior in sheep.


Asunto(s)
Glándula Pineal , Transcriptoma , Ovinos/genética , Animales , Masculino , RNA-Seq/veterinaria , Hipotálamo/metabolismo , Oveja Doméstica , Fenotipo , Perfilación de la Expresión Génica/veterinaria
2.
J Anim Sci ; 100(5)2022 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-35552417

RESUMEN

Low birth weight (LBW) is associated with metabolic disorders in early life. While dietary l-tryptophan (Trp) can ameliorate postprandial plasma triglycerides (TG) disposal in LBW piglets, the genetic and biological basis underlying Trp-caused alterations in lipid metabolism is poorly understood. In this study, we collected 24 liver samples from 1-mo-old LBW and normal birth weight (NBW) piglets supplemented with different concentrations of dietary Trp (NBW with 0% Trp, N0; LBW with 0% Trp, L0; LBW with 0.4% Trp, L4; LBW with 0.8% Trp, L8; N = 6 in each group.) and conducted systematic, transcriptome-wide analysis using RNA sequencing (RNA-seq). We identified 39 differentially expressed genes (DEG) between N0 and L0, and genes within "increased dose effect" clusters based on dose-series expression profile analysis, enriched in fatty acid response of gene ontology (GO) biological process (BP). We then identified RNA-binding proteins including SRSF1, DAZAP1, PUM2, PCBP3, IGF2BP2, and IGF2BP3 significantly (P < 0.05) enriched in alternative splicing events (ASE) in comparison with L0 as control. There were significant positive and negative relationships between candidate genes from co-expression networks (including PID1, ANKRD44, RUSC1, and CYP2J34) and postprandial plasma TG concentration. Further, we determined whether these candidate hub genes were also significantly associated with metabolic and cardiovascular traits in humans via human phenome-wide association study (Phe-WAS), and analysis of mammalian orthologs suggests a functional conservation between human and pig. Our work demonstrates that transcriptomic changes during dietary Trp supplementation in LBW piglets. We detected candidate genes and related BP that may play roles on lipid metabolism restoration. These findings will help to better understand the amino acid support in LBW metabolic complications.


Low birth weight (LBW) has been associated with higher rate of mortality and morbidity and the development of metabolic complications, leaving burdens on livestock production and human health care. The feasibility of LBW metabolic restoration via postnatal nutrition compensation has been verified and the role of one of essential amino acids, l-tryptophan (Trp), on rescuing lipid metabolism in LBW was determined, while the underlying molecular mechanism and key gene regulation is little known. Our study was conducted to identify the unique molecular mechanisms between LBW and normal birth weight (NBW), and to identify the metabolic restoration related genes and biological processes after dietary Trp supplementation in LBW piglet model. We found that differentially expressed genes (DEG) between LBW and NBW were related to fatty acid response based on gene ontology enrichment analysis, and LBW piglets supplemented with Trp showed lower postprandial plasma triglycerides (TG) level as NBW, with similar expression feature of lipid metabolism related genes.


Asunto(s)
Suplementos Dietéticos , Triptófano , Animales , Peso al Nacer , Humanos , Mamíferos/metabolismo , Proteínas de Unión al ARN , RNA-Seq/veterinaria , Análisis de Secuencia de ARN/veterinaria , Factores de Empalme Serina-Arginina , Porcinos , Triglicéridos , Triptófano/metabolismo , Triptófano/farmacología
3.
Vet Med Sci ; 8(1): 367-376, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34822225

RESUMEN

The conditions for sex reversal in vertebrate species have been extensively studied, and the results highlighted numerous key factors involved in sex differentiation. However, the transcriptomes in hypothalamic and pituitary tissues from intersex goats have rarely been studied. The aim of this study was to screen candidate genes and signalling pathways related to sex reversal in Huai goats by analyzing gene expression in hypothalamic and pituitary tissues via transcriptome sequencing and bioinformatics analyses. In total, 612 and 139 differentially expressed genes (DEGs) were identified between the intersex and non-intersex groups in the hypothalamus and pituitary, respectively. The DEGs in the hypothalamus and pituitary were significantly enriched in 41 and 16 signalling pathways, respectively, including the calcium signalling pathway, neuroactive ligand-receptor interaction signalling pathway, and oestrogen signalling pathway, which might be related to intersex sex development disorders. A candidate gene from the tachykinin family (TACR1) was significantly enriched in the calcium signalling pathway. Thirty-one DEGs were shared between these two comparisons and were enriched in several acetyl-CoA-related processes and the oestrogen signalling pathway. The results of the real-time PCR analysis show that the transcriptome sequencing results were reliable. The transcriptome data indicate that the regulation of various physiological systems is involved in intersex goat development. Therefore, these results provide helpful data enhancing our understanding of the molecular mechanisms underlying intersex syndrome in goats.


Asunto(s)
Trastornos del Desarrollo Sexual , Enfermedades de las Cabras , Animales , Trastornos del Desarrollo Sexual/genética , Trastornos del Desarrollo Sexual/veterinaria , Perfilación de la Expresión Génica/veterinaria , Cabras/genética , Hipotálamo , RNA-Seq/veterinaria , Transcriptoma
4.
Trop Anim Health Prod ; 53(1): 131, 2021 Jan 19.
Artículo en Inglés | MEDLINE | ID: mdl-33462736

RESUMEN

Taiping chicken is indigenous chickens (Gallus gallus domesticus), which was one of China's excellent poultry species. As the problems caused by the overuse of antibiotics become more and more concern, people begin to look for ways to replace them. Among them, probiotics and their preparations are the research hotspot to replace antibiotics. Probiotics can promote the absorption of nutrients, improve the ratio of meat to meat, resist and prevent diseases, and improve the intestinal tissue morphology. Here, we performed transcriptome profiling of Taiping chicken ileum which was given probiotics by using the Hiseq™ 2500 sequencing platform. A total of 18 genes were differentially expressed in the ileum under control group and probiotics group. Thirteen genes were upregulated with a range of fold change from 1.02 to 8.61, and 5 were downregulated with a range of fold change from - 1.06 to - 2.29. Ten of the differently expressed genes were further validated by qRT-PCR. In addition, Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that these differentially expressed genes were mainly enriched to bile secretion, alpha-linolenic acid metabolism, linoleic acid metabolism, ether lipid metabolism, and arachidonic acid metabolism. This study will help us to understand the role of probiotics in indigenous chicken production and provide theoretical basis for the genetic development of indigenous chickens.


Asunto(s)
Pollos , Íleon , Probióticos , RNA-Seq/veterinaria , Animales , Pollos/genética , Perfilación de la Expresión Génica/veterinaria , Transcriptoma
5.
Theriogenology ; 157: 449-457, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32882647

RESUMEN

Oocytes of better quality and developmental competence are highly demanded, which is affected by many intrinsic and external factors, including environmental pollutants. We have previously demonstrated that 7, 12-dimethylbenz [a]anthracene (DMBA) reduces the developmental competence of porcine oocytes, by desynchronizing nuclear and ooplasmic maturation. However, the underlying molecular mechanism remains obscure. Here we performed single cell RNA-seq to study the transcriptome changes in DMBA-treated porcine MII oocytes, and identified 19 protein-coding genes and 156 novel long non-coding RNAs (lncRNAs) with abundance to be significantly different (P < 0.05), which enriched in signaling pathways such as glycosphingolipid biosynthesis, nicotine addiction, basal transcription factors and nucleotide excision repair. RT-qPCR on oocyte pools confirmed ornithine aminotransferase (Oat) and serine/arginine-rich splicing factor 4 (Srsf4) to be significantly up- and down-regulated, respectively (P < 0.05). Treating porcine COCs with MAPK and PLC pathway inhibitors suppressed DMBA's effects on increasing PB1 extrusion rate. In addition, DMBA co-incubation with 250 µM vitamin C derivative (l-ascorbic acid 2-phosphate sesquimagnesium salt hydrate, AA2P) and 100 µM co-enzyme Q10 (CoQ10) could significantly reduce the DMBA-induced high ROS level, and partially alleviate the DMBA-induced high PB1 rate, whereas the cleavage and blastocyst rates of parthenotes derived from treated mature oocytes remained to be low. Collectively, our findings indicate that single cell RNA-seq can help reveal the dynamics of molecular signaling pathways for porcine oocytes treated by DMBA, and supplement of anti-oxidative reagents could not sufficiently rescue DMBA-induced defects of porcine oocytes.


Asunto(s)
Células del Cúmulo , Oocitos , Animales , Antracenos , Femenino , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oogénesis , RNA-Seq/veterinaria , Porcinos
6.
Fish Shellfish Immunol ; 106: 8-20, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32717323

RESUMEN

Functional ingredients such as Bacillus subtilis are used in aquaculture to improve fish condition, modulate microbiota and promote a healthy intestinal system. However, the underlying mechanisms of grass carp treated with B. subtilis are not fully characterized. This study investigated the gut microbes of grass carp after treated with B. subtilis H2 (106 CFU/mL) and Aeromonas hydrophila (106 CFU/mL). The intestinal flora was found that the dominant bacterial phyla identified in all samples were Proteobacteria, Actinobacteria, Fusobacteria, Bacteroidetes and Acidobacteria. Compared with the control group, the relative abundance of Proteobacteria and Bacteroidetes in B. subtilis group were significantly increased. In addition, the relative abundances of Aeromonas and Shewanella in A. hydrophila group were more than the control group. For the intestinal transcriptomic profiling of the grass carp treated with B. subtilis H2, 824 different expressed genes (DEGs) between the B. subtilis H2 treated and non-treated groups were detected, including 365 up-regulated and 459 down-regulated genes. Six DEGs were randomly selected for further validation by quantitative real-time RT-PCR (qRT-PCR) and the results were consistent with the RNA-seq data. Additionally, eight immunomodulatory genes (IL-4, IL-11, IFN-α, CSF, FOSB, MAPK12b, IGHV3-11 and IGHV3-21) were significantly up-regulated after treated with B. subtilis H2. Furthermore, almost all the lipid metabolism-associated genes were significantly up-regulated after treated with B. subtilis H2 according to the lipid metabolism pathways. Eleven lipid metabolism-associated genes were selected by qRT-PCR, which showed that the expressions of almost all the selected genes were increased, especially Apob-48, ABCG8 and DGAT. Taken together, our results support that B. subtilis could modulate the immune response, fat metabolism and bacterial assembly in the gut of grass carp.


Asunto(s)
Bacillus subtilis/química , Carpas/inmunología , Enfermedades de los Peces/inmunología , Proteínas de Peces/genética , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Metabolismo de los Lípidos , Aeromonas hydrophila/fisiología , Alimentación Animal/análisis , Animales , Bacterias/aislamiento & purificación , ADN Bacteriano/análisis , Dieta/veterinaria , Suplementos Dietéticos/análisis , Relación Dosis-Respuesta a Droga , Enfermedades de los Peces/genética , Proteínas de Peces/inmunología , Microbioma Gastrointestinal , Perfilación de la Expresión Génica/veterinaria , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/veterinaria , Probióticos/administración & dosificación , Probióticos/metabolismo , ARN Ribosómico 16S/análisis , RNA-Seq/veterinaria , Distribución Aleatoria
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