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Comparative Transcriptome Analysis of Sucrose Biosynthesis-Associated Gene Expression Using RNA-Seq at Various Growth Periods in Sugar Beet (Beta vulgaris L.)

  • Baul Yang;Ye-Jin Lee;Dong-Gun Kim;Sang Hoon Kim;Woon Ji Kim;Jae Hoon Kim;So Hyeon Baek;Joon-Woo Ahn;Chang-Hyu Bae;Jaihyunk Ryu
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
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    • pp.63-63
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    • 2023
  • Sugar beet (Beta vulgaris L.) is one of the most important sugar crops and provides up to 30% of the world's sugar production. In this study, we mainly performed RNA-sequencing to obtain identify putative genes involved in biosynthesis pathway of sucrose in sugar beet and comparative transcriptomic analyses in the four developmental stages (50, 90, 160 and 330 days after seedling). As a result of the sugar content analysis, it was increased significantly from 50 to 160 days after seedling (DAS), and then decreased at 330 DAS. On the other hand, the taproot weight, length, and width were increased during all the growth periods. Out of 21,451 genes with expressed value, 21,402 (99.77%) genes had functional descriptions. Among the three comparisons, S1 (50 DAS) vs. S2 (90 DAS), S1 vs. S3 (160 DAS), and S1 vs. S4 (330 DAS), expression profiling of the transcripts was identified 4,991 with differentially expressed genes (DEGs). By comparing the top 20 enriched gene ontology (GO) terms as three comparisons, the top GO terms were commonly confirmed with external encapsulating structure, cell wall, and extracellular regions. In addition, the 38 enriched candidate genes related to sucrose biosynthetic pathway were screened from the entire DEG pool, and the candidate genes might be providing a basic data for further sugar metabolism studies in development of sugar beet taproot.

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중등 과학 수업의 참여구조 사례 연구: '혼성적 의미 창출 공간'의 형성 가능성 탐색 (Case Studies of the Participation Structures in Secondary Science Classrooms: Exploring the Possibility to Develop the 'Space for Hybrid Meaning Making')

  • 유은정;이선경;오필석;신명경;김찬종
    • 한국과학교육학회지
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    • 제28권6호
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    • pp.603-617
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    • 2008
  • 본 연구에서는 과학 교수-학습에 관한 사회문화적 관점을 바탕으로 과학 수업에서 교사와 학생들이 어떻게 의사소통하고 의미를 창출하는지 살펴보았다. 자료는 10학년 과학 수업 4개분이며, 수업 녹화물과 전사본을 분석하였다. 자료 분석은 중등 과학 수업에서 발견되는 참여구조와 혼성적 의미 창출 공간의 형성 가능성을 중심으로 이루어졌다. 연구에서 확인된 참여구조들은 대체로 교사 주도적이었으며, 새로운 의미 창출을 위해 학생들이 능동적으로 담화를 선도하는 경우는 드물었다. 하지만, 몇 가지 참여구조들은 혼성적 의미 창출 공간으로 발전할 가능성을 다소간 내포하고 있음을 논의하였다. 연구 결과를 토대로 보다 바람직한 교육적 담화와 장래 연구에 대한 시사점을 제시하였다.

시베리아 철갑상어(Acipenser baerii) 트랜스페린(Transferrin) 유전자의 발생 배 및 초기 자어에서의 발현 특징 (Expression Characteristics of Transferrin Gene During Embryo Development and Prolarval Ontogeny in Siberian Sturgeon Acipenser baerii)

  • 김은정;남윤권
    • 한국수산과학회지
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    • 제57권1호
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    • pp.41-52
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    • 2024
  • Serotransferrin cDNA from the Siberian sturgeon Acipenser baerii was isolated and its expression patterns during early life intervals were characterized. It contained an ORF encoding a 708-aa-long polypeptide, including a 19-aa signal peptide. Bioinfomatic analysis and 3D modeling indicated a typical bi-lobal structure with conserved iron-coordinating residues. During embryonic development, the potential transition of maternally provisioned transcripts to zygotically de novo transcribed ones occurred around blastula stage. The transferrin mRNA levels peaked at stages responsible for pronephros, heart and erythropoietic component differentiation. After hatching, the transferrin mRNA expression gradually increased at early ontogenic phases (0 to 3 DPH) corresponding to the periods in which prolarvae exhibited increased blood circulation and liver differentiation. The expression decreased at subsequent stages in which prolarvae exhibited benthic movement. The tissue distribution assay indicated liver-predominant expression at fingerling stage. From the microinjection-based challenge with Aeromonas hydrophila at day-0 and day-7, the transcriptional response was modulated toward upregulation, in which the amounts induced at 6, 12 and 24 HPI were greater in prolarvae injected at day-7 than at day-0. Therefore, transferrin plays important roles in both early development and host protective responses to pathogens in the Siberian sturgeon.

Single-cell RNA sequencing reveals the heterogeneity of adipose tissue-derived mesenchymal stem cells under chondrogenic induction

  • Jeewan Chun;Ji-Hoi Moon;Kyu Hwan Kwack;Eun-Young Jang;Saebyeol Lee;Hak Kyun Kim;Jae-Hyung Lee
    • BMB Reports
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    • 제57권5호
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    • pp.232-237
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    • 2024
  • This study investigated how adipose tissue-derived mesenchymal stem cells (AT-MSCs) respond to chondrogenic induction using droplet-based single-cell RNA sequencing (scRNA-seq). We analyzed 37,219 high-quality transcripts from control cells and cells induced for 1 week (1W) and 2 weeks (2W). Four distinct cell clusters (0-3), undetectable by bulk analysis, exhibited varying proportions. Cluster 1 dominated in control and 1W cells, whereas clusters (3, 2, and 0) exclusively dominated in control, 1W, and 2W cells, respectively. Furthermore, heterogeneous chondrogenic markers expression within clusters emerged. Gene ontology (GO) enrichment analysis of differentially expressed genes unveiled cluster-specific variations in key biological processes (BP): (1) Cluster 1 exhibited up-regulation of GO-BP terms related to ribosome biogenesis and translational control, crucial for maintaining stem cell properties and homeostasis; (2) Additionally, cluster 1 showed up-regulation of GO-BP terms associated with mitochondrial oxidative metabolism; (3) Cluster 3 displayed up-regulation of GO-BP terms related to cell proliferation; (4) Clusters 0 and 2 demonstrated similar up-regulation of GO-BP terms linked to collagen fibril organization and supramolecular fiber organization. However, only cluster 0 showed a significant decrease in GO-BP terms related to ribosome production, implying a potential correlation between ribosome regulation and the differentiation stages of AT-MSCs. Overall, our findings highlight heterogeneous cell clusters with varying balances between proliferation and differentiation before, and after, chondrogenic stimulation. This provides enhanced insights into the single-cell dynamics of AT-MSCs during chondrogenic differentiation.

Trypanosoma cruzi Dysregulates piRNAs Computationally Predicted to Target IL-6 Signaling Molecules During Early Infection of Primary Human Cardiac Fibroblasts

  • Ayorinde Cooley;Kayla J. Rayford;Ashutosh Arun;Fernando Villalta;Maria F. Lima;Siddharth Pratap;Pius N. Nde
    • IMMUNE NETWORK
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    • 제22권6호
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    • pp.51.1-51.20
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    • 2022
  • Trypanosoma cruzi, the etiological agent of Chagas disease, is an intracellular protozoan parasite, which is now present in most industrialized countries. About 40% of T. cruzi infected individuals will develop severe, incurable cardiovascular, gastrointestinal, or neurological disorders. The molecular mechanisms by which T. cruzi induces cardiopathogenesis remain to be determined. Previous studies showed that increased IL-6 expression in T. cruzi patients was associated with disease severity. IL-6 signaling was suggested to induce pro-inflammatory and pro-fibrotic responses, however, the role of this pathway during early infection remains to be elucidated. We reported that T. cruzi can dysregulate the expression of host PIWI-interacting RNAs (piRNAs) during early infection. Here, we aim to evaluate the dysregulation of IL-6 signaling and the piRNAs computationally predicted to target IL-6 molecules during early T. cruzi infection of primary human cardiac fibroblasts (PHCF). Using in silico analysis, we predict that piR_004506, piR_001356, and piR_017716 target IL6 and SOCS3 genes, respectively. We validated the piRNAs and target gene expression in T. cruzi challenged PHCF. Secreted IL-6, soluble gp-130, and sIL-6R in condition media were measured using a cytokine array and western blot analysis was used to measure pathway activation. We created a network of piRNAs, target genes, and genes within one degree of biological interaction. Our analysis revealed an inverse relationship between piRNA expression and the target transcripts during early infection, denoting the IL-6 pathway targeting piRNAs can be developed as potential therapeutics to mitigate T. cruzi cardiomyopathies.

TCF4-Targeting miR-124 is Differentially Expressed amongst Dendritic Cell Subsets

  • Sun Murray Han;Hye Young Na;Onju Ham;Wanho Choi;Moah Sohn;Seul Hye Ryu;Hyunju In;Ki-Chul Hwang;Chae Gyu Park
    • IMMUNE NETWORK
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    • 제16권1호
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    • pp.61-74
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    • 2016
  • Dendritic cells (DCs) are professional antigen-presenting cells that sample their environment and present antigens to naïve T lymphocytes for the subsequent antigen-specific immune responses. DCs exist in a range of distinct subpopulations including plasmacytoid DCs (pDCs) and classical DCs (cDCs), with the latter consisting of the cDC1 and cDC2 lineages. Although the roles of DC-specific transcription factors across the DC subsets have become understood, the posttranscriptional mechanisms that regulate DC development are yet to be elucidated. MicroRNAs (miRNAs) are pivotal posttranscriptional regulators of gene expression in a myriad of biological processes, but their contribution to the immune system is just beginning to surface. In this study, our in-house probe collection was screened to identify miRNAs possibly involved in DC development and function by targeting the transcripts of relevant mouse transcription factors. Examination of DC subsets from the culture of mouse bone marrow with Flt3 ligand identified high expression of miR-124 which was able to target the transcript of TCF4, a transcription factor critical for the development and homeostasis of pDCs. Further expression profiling of mouse DC subsets isolated from in vitro culture as well as via ex vivo purification demonstrated that miR-124 was outstandingly expressed in CD24+ cDC1 cells compared to in pDCs and CD172α+ cDC2 cells. These results imply that miR-124 is likely involved in the processes of DC subset development by posttranscriptional regulation of a transcription factor(s).

Yeast Small Ubiquitin-Like Modifier (SUMO) Protease Ulp2 is Involved in RNA Splicing

  • Jeong-Min Park;Seungji Choi;Dong Kyu Choi;Hyun-Shik Lee;Dong-Hyung Cho;Jungmin Choi;Hong-Yeoul Ryu
    • 한국발생생물학회지:발생과생식
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    • 제28권2호
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    • pp.47-54
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    • 2024
  • In eukaryotes, RNA splicing, an essential biological process, is crucial for precise gene expression. Inaccurate RNA splicing can cause aberrant mRNA production, disrupting protein synthesis. To regulate splicing efficiency, some splicing factors are reported to undergo Ubiquitin-like Modifier (SUMO)ylation. Our data indicate that in Saccharomyces cerevisiae, the SUMO protease, Ulp2, is involved in splicing. In the ulp2Δ mutant, some ribosomal protein (RP) transcripts exhibited a significant increase in the levels of intron-containing pre-mRNA because of improper splicing. Moreover, we confirmed Ulp2 protein binding to the intronic regions of RP genes. These findings highlight a critical Ulp2 role in RP transcript splicing.

CircZNF609 Aggravated Myocardial Ischemia Reperfusion Injury via Mediation of miR-214-3p/PTGS2 Axis

  • Wen-Qiang Tang;Feng-Rui Yang;Ke-Min Chen;Huan Yang;Yu Liu;Bo Dou
    • Korean Circulation Journal
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    • 제52권9호
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    • pp.680-696
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    • 2022
  • Background and Objectives: Circular RNAs were known to play vital role in myocardial ischemia reperfusion injury (MIRI), while the role of CircZNF609 in MIRI remains unclear. This study was aimed to investigate the function of CircZNF609 in MIRI. Methods: Hypoxia/reoxygenation (H/R) model was established to mimic MIRI in vitro. Quantitative polymerase chain reaction was performed to evaluate gene transcripts. Cellular localization of CircZNF609 and miR-214-3p were visualized by fluorescence in situ hybridization. Cell proliferation was determined by CCK-8. TUNEL assay and flow cytometry were applied to detect apoptosis. Lactate dehydrogenase was determined by commercial kit. ROS was detected by DCFH-DA probe. Direct interaction of indicated molecules was determined by RIP and dual luciferase assays. Western blot was used to quantify protein levels. In vivo model was established to further test the function of CircZNF609 in MIRI. Results: CircZNF609 was upregulated in H/R model. Inhibition of CircZNF609 alleviated H/R induced apoptosis, ROS generation, restored cell proliferation in cardiomyocytes and human umbilical vein endothelial cells. Mechanically, CircZNF609 directly sponged miR-214-3p to release PTGS2 expression. Functional rescue experiments showed that miR-214-3p/PTGS2 axis was involved in the function of circZNG609 in H/R model. Furthermore, data in mouse model revealed that knockdown of CircZNF609 significantly reduced the area of myocardial infarction and decreased myocardial cell apoptosis. Conclusions: CircZNF609 aggravated the progression of MIRI via targeting miR-214-3p/PTGS2 axis, which suggested CircZNF609 might act as a vital modulator in MIRI.

Molecular cloning, identification, transcriptional analysis, and silencing of enolase on the life cycle of Haemaphysalis longicornis (Acari, Ixodidae) tick

  • Md. Samiul Haque;Md. Khalesur Rahman;Mohammad Saiful Islam;Myung-Jo You
    • Parasites, Hosts and Diseases
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    • 제62권2호
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    • pp.226-237
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    • 2024
  • Ticks, blood-sucking ectoparasites, spread diseases to humans and animals. Haemaphysalis longicornis is a significant vector for tick-borne diseases in medical and veterinary contexts. Identifying protective antigens in H. longicornis for an anti-tick vaccine is a key tick control strategy. Enolase, a multifunctional protein, significantly converts D-2-phosphoglycerate and phosphoenolpyruvate in glycolysis and gluconeogenesis in cell cytoplasm. This study cloned a complete open reading frame (ORF) of enolase from the H. longicornis tick and characterized its transcriptional and silencing effect. We amplified the full-length cDNA of the enolase gene using rapid amplification of cDNA ends. The complete cDNA, with an ORF of 1,297 nucleotides, encoded a 432-amino acid polypeptide. Enolase of the Jeju strain H. longicornis exhibited the highest sequence similarity with H. flava (98%), followed by Dermacentor silvarum (82%). The enolase motifs identified included N-terminal and C-terminal regions, magnesium binding sites, and several phosphorylation sites. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that enolase mRNA transcripts were expressed across all developmental stages of ticks and organs such as salivary gland and midgut. RT-PCR showed higher transcript levels in syn-ganglia, suggesting that synganglion nerves influence enolase's role in tick salivary glands. We injected enolase double-stranded RNA into adult unfed female ticks, after which they were subsequently fed with normal unfed males until they spontaneously dropped off. RNA interference significantly (P<0.05) reduced feeding and reproduction, along with abnormalities in eggs (no embryos) and hatching. These findings suggest enolase is a promising target for future tick control strategies.

마우스 대식 세포주 RAW264.7에 있어서 LPS처리에 의해 유도되는 염증반응에 대한 식용피(Echinochloa crus-galli var. frumentacea)의 저해효과 (Anti-inflammatory Effect of Flavonoids Kaempferol and Biochanin A-enriched Extract of Barnyard Millet (Echinochloa crus-galli var. frumentacea) Grains in LPS-stimulated RAW264.7 Cells)

  • 이지영;전도연;윤영호;고지연;우관식;우미희;김영호
    • 생명과학회지
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    • 제24권11호
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    • pp.1157-1167
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    • 2014
  • 잡곡은 대사성 질환의 예방과 치료에 우수한 기능성 식품소재로 인식되고 있다. 국내산 잡곡들(기장, 황금찰수수, 노랑차조, 율무 및 식용피)을 대상으로 항염증 활성을 비교조사하고자, 이들 잡곡의 80% 에탄올 추출물을 이용하여 마우스 대식세포주 RAW264.7을 지질다당류(LPS)로 자극할 때 생성되는 염증 매개인자인 산화질소(NO)의 생성에 미치는 저해능을 조사하였다. 그 결과, 식용피(Echinochloa crus-galli var. frumentacea) 유래의 에탄올 추출물이 LPS에 의해 유도되는 NO의 생성을 저해하는 항염증 활성에 있어서 가장 우수함을 확인하였다. 식용피의 에탄올 추출물로부터 항염증 관련 성분을 규명하기 위해, 80%에탄올 추출물을 물에 녹인 후 n-hexane, methylene chloride (MC), ethyl acetate (EtOAc), n-butanol (BuOH)과 물 분획들로 단계별 분획하고, 각 분획의 항염증 활성을 조사하였다. RAW264.7 세포에 MC 분획($100{\mu}g/ml$)을 전처리하였을 때, LPS처리에 의해 유도되는 iNOS, COX-2, 그리고 친염증성 사이토카인들(IL-$1{\beta}$, IL-6, 및 TNF-${\alpha}$)의 발현이 현저하게 저해되는 것으로 나타났다. 또한, LPS처리에 의해 유도되는 p38MAPK, JNK 및 ERK의 활성화의 경우도 MC 분획에 의해 농도-의존적으로 저해되는 것으로 나타났다. HPLC분석을 통해, 식용피 유래 MC 분획의 항염증 활성은 식용피의 MC 분획 속의 주요성분인 kaempferol과 biochanin A에 기인함을 확인하였다. 이상의 연구 결과는 식용피 및 식용피의 MC 분획이 염증성 질환과 이와 관련된 대사성 질환의 예방과 예후개선에 유리한 기능성 식품소재 개발에 활용될 수 있음을 시사한다