• Title/Summary/Keyword: Transcriptome Sequencing

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Identification of G Protein Coupled Receptors Expressed in Fat Body of Plutella Xylostella in Different Temperature Conditions (온도 차이에 따른 배추좀나방 유충 지방체에서 발현되는 G 단백질 연관 수용체의 동정)

  • Kim, Kwang Ho;Lee, Dae-Weon
    • Korean Journal of Environmental Agriculture
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    • v.40 no.1
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    • pp.1-12
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    • 2021
  • BACKGROUND: G protein-coupled receptors (GPCRs) are widely distributed in various organisms. Insect GPCRs shown as in vertebrate GPCRs are membrane receptors that coordinate or involve in various physiological processes such as learning/memory, development, locomotion, circadian rhythm, reproduction, etc. This study aimed to identify GPCRs expressed in fat body and compare the expression pattern of GPCRs in different temperature conditions. METHODS AND RESULTS: To identify GPCRs genes and compare their expression in different temperature conditions, total RNAs of fat body in Plutella xylostella larva were extracted and the transcriptomes have been analyzed via next generation sequencing method. From the fat body transcriptomes, genes that belong to GPCR Family A, B, and F were identified such as opsin, gonadotropin-releasing hormone receptor, neuropeptide F (NPF) receptor, muthuselah (Mth), diuretic hormone receptor, frizzled, etc. Under low temperature, expressions of GPCRs such as C-C chemokine receptor (CCR), opsin, prolactin-releasing peptide receptor, substance K receptor, Mth-like receptor, diuretic hormone receptor, frizzled and stan were higher than those at 25℃. They are involved in immunity, feeding, movement, odorant recognition, diuresis, and development. In contrast to the control (25℃), at high temperature GPCRs including CCR, gonadotropin-releasing hormone receptor, moody, NPF receptor, neuropeptide B1 receptor, frizzled and stan revealed higher expression whose biological functions are related to immunity, blood-brain barrier formation, feeding, learning, and reproduction. CONCLUSION: Transcriptome of fat body can provide understanding the pools of GPCRs. Identifications of fat body GPCRs may contribute to develop new targets for the control of insect pests.

Transcriptome-wide analysis reveals gluten-induced suppression of small intestine development in young chickens

  • Darae, Kang;Donghyun, Shin;Hosung, Choe;Doyon, Hwang;Andrew Wange, Bugenyi;Chong-Sam, Na;Hak-Kyo, Lee;Jaeyoung, Heo;Kwanseob, Shim
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.752-769
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    • 2022
  • Wheat gluten is an increasingly common ingredient in poultry diets but its impact on the small intestine in chicken is not fully understood. This study aimed to identify effects of high-gluten diets on chicken small intestines and the variation of their associated transcriptional responses by age. A total of 120 broilers (Ross Strain) were used to perform two animal experiments consisting of two gluten inclusion levels (0% or 25%) by bird's age (1 week or 4 weeks). Transcriptomics and histochemical techniques were employed to study the effect of gluten on their duodenal mucosa using randomly selected 12 broilers (3 chicks per group). A reduction in feed intake and body weight gain was found in the broilers fed a high-gluten containing diet at both ages. Histochemical photomicrographs showed a reduced villus height to crypt depth ratio in the duodenum of gluten-fed broilers at 1 week. We found mainly a significant effect on the gene expression of duodenal mucosa in gluten-fed broilers at 1 week (289 differentially expressed genes [DEGs]). Pathway analyses revealed that the significant DEGs were mainly involved in ribosome, oxidative phosphorylation, and peroxisome proliferator-activated receptor (PPAR) signaling pathways. These pathways are involved in ribosome protein biogenesis, oxidative phosphorylation and fatty acid metabolism, respectively. Our results suggest a pattern of differential gene expression in these pathways that can be linked to chronic inflammation, suppression of cell proliferation, cell cycle arrest and apoptosis. And via such a mode of action, high-gluten inclusion levels in poultry diets could lead to the observed retardation of villi development in the duodenal mucosa of young broiler chicken.

Momordicine I alleviates isoproterenol-induced cardiomyocyte hypertrophy through suppression of PLA2G6 and DGK-ζ

  • Hongming Li;Yumei Qiu;Mengdie Xie;Changsheng Ouyang;Xiaoyun Ding;Hao Zhang;Wei Dong;Yinhua Xiong;Xilan Tang
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.1
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    • pp.75-84
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    • 2023
  • This study aimed to observe the protective effect of momordicine I, a triterpenoid compound extracted from momordica charantia L., on isoproterenol (ISO)-induced hypertrophy in rat H9c2 cardiomyocytes and investigate its potential mechanism. Treatment with 10 μM ISO induced cardiomyocyte hypertrophy as evidenced by increased cell surface area and protein content as well as pronounced upregulation of fetal genes including atrial natriuretic peptide, βmyosin heavy chain, and α-skeletal actin; however, those responses were markedly attenuated by treatment with 12.5 ㎍/ml momordicine I. Transcriptome experiment results showed that there were 381 and 447 differentially expressed genes expressed in comparisons of model/control and momordicine I intervention/model, respectively. GO enrichment analysis suggested that the anti-cardiomyocyte hypertrophic effect of momordicine I may be mainly associated with the regulation of metabolic processes. Based on our transcriptome experiment results as well as literature reports, we selected glycerophospholipid metabolizing enzymes group VI phospholipase A2 (PLA2G6) and diacylglycerol kinase ζ (DGK-ζ) as targets to further explore the potential mechanism through which momordicine I inhibited ISO-induced cardiomyocyte hypertrophy. Our results demonstrated that momordicine I inhibited ISO-induced upregulations of mRNA levels and protein expressions of PLA2G6 and DGK-ζ. Collectively, momordicine I alleviated ISO-induced cardiomyocyte hypertrophy, which may be related to its inhibition of the expression of glycerophospholipid metabolizing enzymes PLA2G6 and DGK-ζ

Transcriptome Profiling Identifies Genes of Waterlogging-Tolerant and -Sensitive Rapeseeds Differentially Respond to Waterlogging Stress at the Flowering Stage

  • Ji-Eun Lee;Da-Hee An;Kwang-Soo Kim;Young-Lok Cha;Dong-Chil Chang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.229-229
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    • 2022
  • Rapeseed is a crop that is waterlogging sensitive, and it is necessary to breed waterlogging tolerance varieties. Our study presents the comparative transcriptome changes in two rapeseed lines, i.e., waterlogging-tolerant (tJ8634-B-30,) and - sensitive ('EMS26') lines under control and waterlogging stress treatments at the flowering stage. RNA-sequencing analysis revealed 13,279 differentially expressed genes (DEGs) for 'J8634-B-30' and 8,682 DEGs for 'EMS26' under waterlogging stress condition compared to control. Among DEGs of 'J8634-B-30', 6,818 were up-regulated and 6,461 were down-regulated. On the other hand, among the DEGs of 'EMS26', the number of down-regulated genes (5,240) were higher than that of up-regulated genes (3,442). Gene ontology enrichment analysis showed that DEGs related to glucan metabolic, cell wall, and oxidoreductase activity were significantly changed in 'J8634-B-30'. Kyoto Encyclopedia of Genes and Genomes (KEGG)-based analysis in 'J8634-B-30' identified up-regulated DEGs being involved in MAPK signaling pathways. In addition, the DEGs belonging to mechanisms responding to waterlogging stress, i.e., plant hormones, carbon metabolism, Reactive oxygen species (ROS), Nitric oxide (NO) etc. were compared in rapeseed lines. Several DEGs including ethylene-responsive transcription factor (ERF), constitutive triple response (CTR) (in ethylene signaling pathway), monodehydroascorbate Reductase (MDAR), NADPH oxidase (in ROS pathway), cytochrome c oxidase assembly protein (COX) (in NO pathway) up-regulated in 'J8634-B-30'. These outcomes provided the valuable information for further exploring the genetic mechanism of waterlogging tolerance in rapeseed.

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Transcriptome profiling of rubber tree (Hevea brasiliensis) discovers candidate regulators of the cold stress response

  • Gong, Xiao-Xiao;Yan, Bing-Yu;Hu, Jin;Yang, Cui-Ping;Li, Yi-Jian;Liu, Jin-Ping;Liao, Wen-Bin
    • Genes and Genomics
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    • v.40 no.11
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    • pp.1181-1197
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    • 2018
  • Tropical plant rubber tree (Hevea brasiliensis) is the sole source of commercial natural rubber and low-temperature stress is the most important limiting factor for its cultivation. To characterize the gene expression profiles of H. brasiliensis under the cold stress and discover the key cold stress-induced genes. Three cDNA libraries, CT (control), LT2 (cold treatment at $4^{\circ}C$ for 2 h) and LT24 (cold treatment at $4^{\circ}C$ for 24 h) were constructed for RNA sequencing (RNA-Seq) and gene expression profiling. Quantitative real time PCR (qRT-PCR) was conducted to validate the RNA-Seq and gene differentially expression results. A total of 1457 and 2328 differentially expressed genes (DEGs) in LT2 and LT24 compared with CT were respectively detected. Most significantly enriched KEGG pathways included flavonoid biosynthesis, phenylpropanoid biosynthesis, plant hormone signal transduction, cutin, suberine and wax biosynthesis, Pentose and glucuronate interconversions, phenylalanine metabolism and starch and sucrose metabolism. A total of 239 transcription factors (TFs) were differentially expressed following 2 h or/and 24 h of cold treatment. Cold-response transcription factor families included ARR-B, B3, BES1, bHLH, C2H, CO-like, Dof, ERF, FAR1, G2-like, GRAS, GRF, HD-ZIP, HSF, LBD, MIKC-MADS, M-type MADS, MYB, MYB-related, NAC, RAV, SRS, TALE, TCP, Trihelix, WOX, WRKY, YABBY and ZF-HD. The genome-wide transcriptional response of rubber tree to the cold treatments were determined and a large number of DEGs were characterized including 239 transcription factors, providing important clues for further elucidation of the mechanisms of cold stress responses in rubber tree.

Anti-proliferative Properties of p-Coumaric Acid in SNU-16 Gastric Cancer Cells (SNU-16 위암 세포주에서 p-coumaric acid의 세포성장 억제 효과)

  • Jang, Mi Gyeong;Ko, Hee Chul;Kim, Se-Jae
    • Journal of Life Science
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    • v.29 no.7
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    • pp.809-816
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    • 2019
  • The ubiquitous plant metabolite p-coumaric acid (p-CA) has antioxidant and anti-inflammatory properties, but its anti-cancer activity has not been established in gastric cancer cell lines. In this study, we investigated the effects of p-CA on the proliferation and transcriptome profile of SNU16 gastric cancer cells. Treatment with p-CA induced apoptosis of the SNU-16 cells by regulating the expression of pro-apoptotic and anti-apoptotic proteins, such as Bcl-2, poly (ADP-ribose) polymerase (PARP), Bax, procaspase-3, and cleaved-caspase-3. The genes differentially expressed in response to p-CA treatment of the SNU-16 cells were identified by RNA sequencing analysis. Genes regulated by p-CA were involved mainly in the inflammatory response, apoptotic processes, cell cycle, and immune response. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that the phosphatidylinositol-3-kinase-Akt and cancer signaling pathways were altered by p-CA. Protein-protein interaction (PPI) network analysis also revealed that p-CA treatment was correlated with differential expression of genes associated with the inflammatory response and cancer. Collectively, these results suggest that p-CA has potential utility in gastric cancer prevention.

Protective effects of Sasa quelpaertensis Leaf Residue Extract against Potassium Oxonate-induced Hyperuricemia in Mice (생쥐에서 제주조릿대 잎 잔사 추출물의 고요산 혈증 저감 효과)

  • Jang, Mi Gyeong;Song, Hana;Lee, Ju Yeop;Ko, Hee Chul;Hur, Sung-Pyo;Kim, Se Jae
    • Journal of Life Science
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    • v.29 no.1
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    • pp.37-44
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    • 2019
  • Leaves of Sasa quelpaertensis Nakai are used in folk medicine for their anti-inflammatory, antipyretic, and diuretic properties. To ensure efficient utilization of S. quelpaertensis leaf, we previously reported a preparation method for phytochemical-rich extract (PRE) using the leaf residue, which was produced after hot water extraction. This study was undertaken to evaluate the hypouricemic potential of S. quelpaertensis leaf PRE in potassium oxonate (PO)-induced hyperuricemic mice. The administration of PRE significantly reduced serum uric acid (UA), blood urea nitrogen (BUN), and serum creatinine levels and increased urine UA and creatinine levels in the PO-induced hyperuricemic mice. It also reduced liver UA levels and xanthine oxidase (XA) activity. A histological analysis revealed that PRE administration protected against PO-induced liver damage, pointing to anti-inflammatory and cytoprotective effects in PO-induced hyperuricemic mice. We analyzed the transcriptome response to PRE administration in PO-induced hyperuricemic mice using RNA sequencing (RNA-Seq) in kidney tissues. The administration of PRE mainly enriched genes involved in mediating immune and inflammatory responses and the metabolic pathway. A Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the metabolic pathway, purine metabolism, and antibody biosynthesis were the major pathways altered in the PRE and PO groups. These results suggest a potential role for PRE in the prevention and treatment of hyperuricemia with inflammation.

Potential Risk of Choline Alfoscerate on Isoflurane-Induced Toxicity in Primary Human Astrocytes

  • Hyun Jung Lee;Hye Rim Cho;Minji Bang;Yeo Song Lee; Youn Jin Kim; Kyuha Chong
    • Journal of Korean Neurosurgical Society
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    • v.67 no.4
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    • pp.418-430
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    • 2024
  • Objective : Isoflurane, a widely used common inhalational anesthetic agent, can induce brain toxicity. The challenge lies in protecting neurologically compromised patients from neurotoxic anesthetics. Choline alfoscerate (L-α-Glycerophosphorylcholine, α-GPC) is recognized for its neuroprotective properties against oxidative stress and inflammation, but its optimal therapeutic window and indications are still under investigation. This study explores the impact of α-GPC on human astrocytes, the most abundant cells in the brain that protect against oxidative stress, under isoflurane exposure. Methods : This study was designed to examine changes in factors related to isoflurane-induced toxicity following α-GPC administration. Primary human astrocytes were pretreated with varying doses of α-GPC (ranging from 0.1 to 10.0 µM) for 24 hours prior to 2.5% isoflurane exposure. In vitro analysis of cell morphology, water-soluble tetrazolium salt-1 assay, quantitative real-time polymerase chain reaction, proteome profiler array, and transcriptome sequencing were conducted. Results : A significant morphological damage to human astrocytes was observed in the group that had been pretreated with 10.0 mM of α-GPC and exposed to 2.5% isoflurane. A decrease in cell viability was identified in the group pretreated with 10.0 µM of α-GPC and exposed to 2.5% isoflurane compared to the group exposed only to 2.5% isoflurane. Quantitative real-time polymerase chain reaction revealed that mRNA expression of heme-oxygenase 1 and hypoxia-inducible factor-1α, which were reduced by isoflurane, was further suppressed by 10.0 µM α-GPC pretreatment. The proteome profiler array demonstrated that α-GPC pretreatment influenced a variety of factors associated with apoptosis induced by oxidative stress. Additionally, transcriptome sequencing identified pathways significantly related to changes in isoflurane-induced toxicity caused by α-GPC pretreatment. Conclusion : The findings suggest that α-GPC pretreatment could potentially enhance the vulnerability of primary human astrocytes to isoflurane-induced toxicity by diminishing the expression of antioxidant factors, potentially leading to amplified cell damage.

Current status and prospects of citrus genomics (감귤 유전체 연구 동향 및 전망)

  • Kim, Ho Bang;Lim, Sanghyun;Kim, Jae Joon;Park, Young Cheol;Yun, Su-Hyun;Song, Kwan Jeong
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.326-335
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    • 2015
  • Citrus is an economically important fruit tree with the largest amount of fruit production in the world. It provides important nutrition such as vitamin C and other health-promoting compounds including its unique flavonoids for human health. However, it is classified into the most difficult crops to develop new cultivars through conventional breeding approaches due to its long juvenility and some unique reproductive biological features such as gamete sterility, nucellar embryony, and high level of heterozygosity. Due to global warming and changes in consumer trends, establishing a systematic and efficient breeding programs is highly required for sustainable production of high quality fruits and diversification of cultivars. Recently, reference genome sequences of sweet orange and clementine mandarin have been released. Based on the reference whole-genome sequences, comparative genomics, reference-guided resequencing, and genotyping-by-sequencing for various citrus cultivars and crosses could be performed for the advance of functional genomics and development of traits-related molecular markers. In addition, a full understanding of gene function and gene co-expression networks can be provided through combined analysis of various transcriptome data. Analytic information on whole-genome and transcriptome will provide massive data on polymorphic molecular markers such as SNP, INDEL, and SSR, suggesting that it is possible to construct integrated maps and high-density genetic maps as well as physical maps. In the near future, integrated maps will be useful for map-based precise cloning of genes that are specific to citrus with major agronomic traits to facilitate rapid and efficient marker-assisted selection.

Transcriptome Profiling and Characterization of Drought-Tolerant Potato Plant (Solanum tuberosum L.)

  • Moon, Ki-Beom;Ahn, Dong-Joo;Park, Ji-Sun;Jung, Won Yong;Cho, Hye Sun;Kim, Hye-Ran;Jeon, Jae-Heung;Park, Youn-il;Kim, Hyun-Soon
    • Molecules and Cells
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    • v.41 no.11
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    • pp.979-992
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    • 2018
  • Potato (Solanum tuberosum L.) is the third most important food crop, and breeding drought-tolerant varieties is vital research goal. However, detailed molecular mechanisms in response to drought stress in potatoes are not well known. In this study, we developed EMS-mutagenized potatoes that showed significant tolerance to drought stress compared to the wild-type (WT) 'Desiree' cultivar. In addition, changes to transcripts as a result of drought stress in WT and drought-tolerant (DR) plants were investigated by de novo assembly using the Illumina platform. One-week-old WT and DR plants were treated with -1.8 Mpa polyethylene glycol-8000, and total RNA was prepared from plants harvested at 0, 6, 12, 24, and 48 h for subsequent RNA sequencing. In total, 61,100 transcripts and 5,118 differentially expressed genes (DEGs) displaying up- or down-regulation were identified in pairwise comparisons of WT and DR plants following drought conditions. Transcriptome profiling showed the number of DEGs with up-regulation and down-regulation at 909, 977, 1181, 1225 and 826 between WT and DR plants at 0, 6, 12, 24, and 48 h, respectively. Results of KEGG enrichment showed that the drought tolerance mechanism of the DR plant can mainly be explained by two aspects, the 'photosynthetic-antenna protein' and 'protein processing of the endoplasmic reticulum'. We also divided eight expression patterns in four pairwise comparisons of DR plants (DR0 vs DR6, DR12, DR24, DR48) under PEG treatment. Our comprehensive transcriptome data will further enhance our understanding of the mechanisms regulating drought tolerance in tetraploid potato cultivars.