scholarly journals Transcriptome Profiling of the Retained Fetal Membranes—An Insight in the Possible Pathogenesis of the Disease

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 675
Author(s):  
Joanna Jaworska ◽  
Katarzyna Ropka-Molik ◽  
Katarzyna Piórkowska ◽  
Tomasz Szmatoła ◽  
Ilona Kowalczyk-Zięba ◽  
...  
Keyword(s):  
Post Partum ◽  
Transcriptome Profiling ◽  
Biological Pathways ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Fetal Membranes ◽  
Gene Ontology Analysis ◽  
Rna Seq ◽  
Matrix Metabolism ◽  
Profound Insight

Retained fetal membranes (RFM) is one of the most common post-partum diseases of a complex etiology. Moreover, its pathogenesis is still not elucidated. Detailed transcriptomic analysis of physiological and retained placenta may bring profound insight in the pathogenesis of the disease. The aim of the study was to compare the transcriptome of the retained and physiologically released placenta as well as biological pathways and processes in order to determine the possible pathogenesis of the disease. Samples of the endometrium and the allantochorion were taken within 2 h after parturition from control mares (n = 3) and mares with RFM (n = 3). RNA sequencing was performed with the use of all samples and mRNA expression of chosen genes was validated with Real Time PCR. Analysis of RNA-seq identified 487 differentially expressed genes in the allantochorion and 261 in the endometrium of control and RFM mares (p < 0.0001). Within genes that may be important in the release of fetal membranes and were differentially expressed, our report pinpointed BGN, TIMP1, DRB, CD3E, C3, FCN3, CASP3, BCL2L1. Gene ontology analysis showed possible processes which were altered in RFM that are apoptosis, inflammatory-related processes, and extracellular matrix metabolism and might be involved in the pathogenesis of RFM. This is the first report on the transcriptome of RFM and physiologically released placenta in mares.

Identification of Differentially Expressed and Spliced Genes with RNA Sequencing Analysis of CLL Specimens

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4582-4582
Author(s):  
Wei Liao ◽  
Gwen Jordaan ◽  
Artur Jaroszewicz ◽  
Matteo Pellegrini ◽  
Sanjai Sharma
Keyword(s):  
B Cells ◽  
Rna Sequencing ◽  
Differentially Expressed Genes ◽  
Fold Change ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Sequencing Analysis ◽  
Rna Seq ◽  
Mrna Isoforms ◽  
Core Set

Abstract Abstract 4582 High throughput sequencing of cellular mRNA provides a comprehensive analysis of the transcriptome. Besides identifying differentially expressed genes in different cell types, it also provides information of mRNA isoforms and splicing alterations. We have analyzed two CLL specimens and a normal peripheral blood B cells mRNA by this approach and performed data analysis to identify differentially expressed and spliced genes. The result showed CLLs specimens express approximately 40% more transcripts compared to normal B cells. The FPKM data (fragment per kilobase of exon per million) revealed a higher transcript expression on chromosome 12 in CLL#1 indicating the presence of trisomy 12, which was confirmed by fluorescent in-situ hybridization assay. With a two-fold change in FPKM as a cutoff and a p value cutoff of 0.05 as compared to the normal B cell control, 415 genes and 174 genes in CLL#1 and 676 and 235 genes in CLL#2 were up and downregulated or differentially expressed. In these two CLL specimens, 45% to 75% of differentially expressed genes are common to both the CLL specimens indicating that genetically disparate CLL specimens have a high percentage of a core set of genes that are potentially important for CLL biology. Selected differentially expressed genes with increased expression (selectin P ligand, SELPLG, and adhesion molecule interacts with CXADR antigen 1, AMICA) and decreased (Fos, Jun, CD69 and Rhob) expression based on the FPKM from RNA-sequencing data were also analyzed in additional CLL specimens by real time PCR analysis. The expression data from RNA-seq closely matches the fold-change in expression as measured by RT-PCR analysis and confirms the validity of the RNA-seq analysis. Interestingly, Fos was identified as one of the most downregulated gene in CLL. Using the Cufflinks and Cuffdiff software, the splicing patterns of genes in CLL specimens and normal B cells were analyzed. Approximately, 1100 to 1250 genes in the two CLL specimens were significantly differentially spliced as compared to normal B cells. In this analysis as well, there is a core set of 800 common genes which are differentially spliced in the two CLL specimens. The RNA-sequencing analysis accurately identifies differentially expressed novel genes and splicing variations that will help us understand the biology of CLL. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Transcriptome profiling provides new insights into florets number difference of inflorescence in lavandula angustifolia

2020 ◽  
Author(s):  
haiping hao ◽  
xiao pei zhu ◽  
Hui Li ◽  
Jian Gong ◽  
Amjad Farooq ◽  
...  
Keyword(s):  
Essential Oil ◽  
Differentially Expressed Genes ◽  
Transcriptome Profiling ◽  
Extraction Methods ◽  
Essential Oil Composition ◽  
Differentially Expressed ◽  
Rna Seq ◽  
Oil Composition ◽  
Aba Metabolism ◽  
Aba Content

Abstract Background Lavender flowers essential oil had been for a variety of therapeutic and cosmetic purposes, and had been popular for centuries. The previous studies of lavender mainly focused on essential oil composition and extraction methods, ignoring the factors which affected the production of essential oils, such as the floret number. This study aims to get a deeper insight into florets number difference mechanism. Results Hormone profile showed positive correlation between ABA content and the number of florets while IAA was negatively correlated. RNA-Seq results showed that 2848 differentially expressed genes screened by comparing different florets samples in one plant. By analyzing dynamic changes of differentially expressed genes, many potentially interesting genes were identified that encoded putative regulators or key components of ABA metabolism and signaling transduction, such as NCED, PYL, PP2C, SnRK2. These genes were highlighted to reveal their importance in regulation of florets numbers. Conclusions 1. The different ABA concentrations lead to florets difference in the Lavandula angustifolia “JX-2” clusters; 2. ABA may affect the florets number by regulating IAA transport and accumulation. The results will be useful for a better understanding of the molecular mechanism on florets number difference that could be laid the foundation for molecular breeding of muti-flortes varieties.

scholarly journals Transcriptome Analysis Reveals Key Genes and Pathways Associated with Mummify Piglets

Genome ◽  
2021 ◽  
Author(s):  
Shujie Wang ◽  
Pingxian Wu ◽  
Kai Wang ◽  
Xiang Ji ◽  
Dong Chen ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Transcriptome Profiling ◽  
Differentially Expressed ◽  
Reproductive Capacity ◽  
Signal Pathways ◽  
Comparative Transcriptome ◽  
Rna Seq ◽  
Promising Candidate ◽  
Pork Consumption ◽  
Differential Genes

China is the country with the largest pork consumption in the world. However, the incidence of high mummify piglets (3-5%) is one of the important factors that cause the slow improvement of pig reproductive capacity, and the genetic mechanism is still unclear. This study aimed to identify candidate genes related to high mummify piglets. RNA-seq technology was used to comparative transcriptome profiling of blood from high piglets mummified and healthy sow at different stages of pregnancy (35d, 56d, 77d and 98d). A total of 137 to 420 DEGs were detected in each stage. Seven differentially expressed genes were significantly differentially expressed at various stages. IL-9R, TLR8, ABLIM3, FSH-α, ASCC1, PRKCZ, and GCK may play an important role in course of mummify piglets. The differential genes we identified between the groups were mainly enriched in immune and inflammation regulation, and others were mainly enriched in reproduction. Considering the function of candidate genes, IL-9R and TLR8 were suggested as the most promising candidate genes involved in mummify piglet traits. We speculate that during pregnancy, it may be the combined effects of the above-mentioned inflammation, immune response, and reproduction-related signal pathways that affect the occurrence of mummifying piglets, and further affect pig reproduction.

scholarly journals Transcriptome analysis of two Pogostemon cablin chemotypes reveals genes related to patchouli alcohol biosynthesis

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12025
Author(s):  
Wuping Yan ◽  
Zhouchen Ye ◽  
Shijia Cao ◽  
Guanglong Yao ◽  
Jing Yu ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Differentially Expressed ◽  
Perennial Herb ◽  
Pcr Analysis ◽  
Biosynthetic Pathways ◽  
Rna Seq ◽  
Patchouli Alcohol ◽  
Pogostemon Cablin ◽  
Polymerase Chain ◽  
Distribution Frequency

Pogostemon cablin, a medicinally and economically important perennial herb, is cultivated around the world due to its medicinal and aromatic properties. Different P. cablin cultivars exhibit different morphological traits and patchouli oil components and contents (especially patchouli alcohol (PA) and pogostone (PO)). According to the signature constituent of the leaf, P. cablin was classified into two different chemotypes, including PA-type and PO-type. To better understand the molecular mechanisms of PA biosynthesis, the transcriptomes of Chinese-cultivated P. cablin cv. PA-type “Nanxiang” (NX) and PO-type “Paixiang” (PX) were analyzed and compared with ribonucleic acid sequencing (RNA-Seq) technology. We obtained a total of 36.83 G clean bases from the two chemotypes, compared them with seven databases and revealed 45,394 annotated unigenes. Thirty-six candidate unigenes participating in the biosynthesis of PA were found in the P. cablin transcriptomes. Overall, 8,390 differentially expressed unigenes were identified between the chemotypes, including 2,467 upregulated and 5,923 downregulated unigenes. Furthermore, six and nine differentially expressed genes (DEGs) were mapped to the terpenoid backbone biosynthetic and sesquiterpenoid and triterpenoid biosynthetic pathways, respectively. One key sesquiterpene synthase gene involved in the sesquiterpenoid and triterpenoid biosynthetic pathways, encoding patchoulol synthase variant 1, was significantly upregulated in NX. Additionally, GC-MS analysis of the two chemotypes in this study showed that the content of PA in NX was significantly higher than that of PX, while the content of PO showed the opposite phenotype. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that the DEG expression tendency was consistent with the transcriptome sequencing results. Overall, 23 AP2/ERF, 13 bHLH, 11 MYB, 11 NAC, three Trihelix, 10 WRKY and three bZIP genes that were differentially expressed may act as regulators of terpenoid biosynthesis. Altogether, 8,314 SSRs were recognized within 6,825 unigenes, with a distribution frequency of 18.32%, among which 1,202 unigenes contained more than one SSR. The transcriptomic characteristics of the two P. cablin chemotypes are comprehensively reported in this study, and these results will contribute to a better understanding of the molecular mechanism of PA biosynthesis. Our transcriptome data also provide a valuable genetic resource for further studies on P. cablin.

scholarly journals Comparative Transcriptome Analysis of a Fan-shaped Inflorescence in Pineapple Using RNA-seq

2020 ◽  
Author(s):  
Tao Xie ◽  
Zhiquan Cai ◽  
Aiping Luan ◽  
Wei Zhang ◽  
Jing Wu ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Soluble Sugar ◽  
Differentially Expressed ◽  
Soluble Solids ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Stem Apex ◽  
Inflorescence Axis ◽  
Flower Stem

Abstract Background: Pineapple plant usually has a capitulum. However, a fan-shaped inflorescence was evolved in an exceptional material, having multiple crown buds. In order to reveal the molecular mechanisms of the formation of the fan-shaped inflorescence, fruit traits and the transcriptional differences between a fan-shaped inflorescence (FI) and a capitulum inflorescence (CI) pineapples were analyzed in the three tissues, i.e., the flower stem apex (FIs and CIs), the base of the inflorescence (FIb and CIb), and the inflorescence axis (FIa and CIa).Results: Except for a clear differentiation of inflorescence morphology, no significant differences in the structure of inflorescence organs and the main nutritional components (soluble solids, soluble sugar, titratable acid, and VC) in fruits were found between the two pineapples. Between the fan- and capitulum-shaped inflorescences, a total of 5370 differentially expressed genes (DEGs) were identified across the three tissues; and 3142, 2526 and 2255 DEGs were found in the flower stem apex, the base of the inflorescence, and the inflorescence axis, respectively. Of these genes, there were 489 overlapping DEGs in all three tissue comparisons. In addition, 5769 DEGs were identified between different tissues within each pineapple. Functional analysis indicated between the two pineapples that 444 transcription factors (TFs) and 206 inflorescence development related genes (IDGs) were differentially expressed in at least one tissue comparison, while 45 TFs and 21 IDGs were overlapped across the 3 tissues. Among the 489 overlapping DEGs in the 3 tissue comparisons between the two pineapples, excluding the IDGs and TFs, 80 of them revealed a higher percentage of involvement in the biological processes relating to response to auxin, and reproductive processes. RNA-seq value and real-time quantitative PCR analysis exhibited the same gene expression patterns in the three tissues. Conclusions: Our result provided novel cues for understanding the molecular mechanisms of the formation of fan-shaped inflorescence in pineapple, making a valuable resource for the study of plant breeding and the speciation of the pineapples.

scholarly journals Physiological Characterization and Transcriptome Analysis of Camellia oleifera Abel. during Leaf Senescence

Forests ◽  
2020 ◽  
Vol 11 (8) ◽  
pp. 812
Author(s):  
Shiwen Yang ◽  
Kehao Liang ◽  
Aibin Wang ◽  
Ming Zhang ◽  
Jiangming Qiu ◽  
...  
Keyword(s):  
Molecular Mechanism ◽  
Leaf Senescence ◽  
Transcriptome Analysis ◽  
Unsaturated Fatty Acids ◽  
De Novo ◽  
Transcriptome Assembly ◽  
Economic Value ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Rna Seq

Camellia (C.) oleifera Abel. is an evergreen small arbor with high economic value for producing edible oil that is well known for its high level of unsaturated fatty acids. The yield formation of tea oil extracted from fruit originates from the leaves, so leaf senescence, the final stage of leaf development, is an important agronomic trait affecting the production and quality of tea oil. However, the physiological characteristics and molecular mechanism underlying leaf senescence of C. oleifera are poorly understood. In this study, we performed physiological observation and de novo transcriptome assembly for annual leaves and biennial leaves of C. oleifera. The physiological assays showed that the content of chlorophyll (Chl), soluble protein, and antioxidant enzymes including superoxide dismutase, peroxide dismutase, and catalase in senescing leaves decreased significantly, while the proline and malondialdehyde concentration increased. By analyzing RNA-Seq data, we identified 4645 significantly differentially expressed unigenes (DEGs) in biennial leaves with most associated with flavonoid and phenylpropanoid biosynthesis and phenylalanine metabolism pathways. Among these DEGs, 77 senescence-associated genes (SAGs) including NOL, ATAF1, MDAR, and SAG12 were classified to be related to Chl degradation, plant hormone, and oxidation pathways. The further analysis of the 77 SAGs based on the Spearman correlation algorithm showed that there was a significant expression correlation between these SAGs, suggesting the potential connections between SAGs in jointly regulating leaf senescence. A total of 162 differentially expressed transcription factors (TFs) identified during leaf senescence were mostly distributed in MYB (myeloblastosis), ERF (Ethylene-responsive factor), WRKY, and NAC (NAM, ATAF1/2 and CUCU2) families. In addition, qRT-PCR analysis of 19 putative SAGs were in accordance with the RNA-Seq data, further confirming the reliability and accuracy of the RNA-Seq. Collectively, we provide the first report of the transcriptome analysis of C. oleifera leaves of two kinds of age and a basis for understanding the molecular mechanism of leaf senescence.

scholarly journals Identification of key pseudogenes in nasopharyngeal carcinoma based on RNA-Seq analysis

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xiujuan Zhang ◽  
Xiaole Song ◽  
Yuting Lai ◽  
Bijun Zhu ◽  
Jiqin Luo ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Cell Line ◽  
Enrichment Analysis ◽  
Arachidonic Acid Metabolism ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Epithelial Cell Line ◽  
Rna Seq ◽  
Multiple Tumor ◽  
East And Southeast Asia

Abstract Background Nasopharyngeal carcinoma (NPC) is a malignant head and neck tumor, and more than 70% of new cases are in East and Southeast Asia. However, association between NPC and pseudogenes playing important roles in genesis of multiple tumor types is still not clear and needs to be investigated. Methods Using RNA-Sequencing (RNA-seq) technology, we analyzed pseudogene expression in 13 primary NPC and 6 recurrent NPC samples as well as their paracancerous counterparts. Quantitative PCR was used to validate the differentially expressed pseudogenes. Results We found 251 differentially expressed pseudogenes including 73 up-regulated and 178 down-regulated ones between primary NPC and paracancerous tissues. Enrichment analysis of gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were conducted to filter out the key pseudogenes. We reported that pseudogenes from cytochrome P450 (CYP) family, such as CYP2F2P, CYP2G1P, CYP4F24P, CYP2B7P and CYP2G2P were significantly down-regulated in NPC compared to paracancerous tissues, while IGHV1OR15–2, IGHV3–11, FCGR1CP and IGHV3–69-1 belonging to Fc gamma receptors were significantly up-regulated. CYP2B7P, CYP2F2P and CYP4F26P were enriched in arachidonic acid metabolism pathway. The qRT-PCR analysis validated the lower expression of pseudogenes CYP2F2P and CYP2B7P in NPC tissues and cell lines compared to paracancerous tissues and normal human nasopharyngeal epithelial cell line. CYP2B7P overexpression weakened migratory and invasive capacity of NPC cell line. Moreover, the expression pattern of those pseudogenes in recurrent NPC tissues was different from the primary NPC. Conclusion This study suggested the role of pseudogenes in tumorigenesis and progression, potentially functioning as therapeutic targets to NPC.

scholarly journals Transcriptome profiling and comparison of Rhinanthus major and Rhinanthus minor reciprocal F1 hybrids during seed stratification and germination

2021 ◽  
Author(s):  
Khaled Mirzaei ◽  
Renate A. Wesselingh
Keyword(s):  
Transcriptome Profiling ◽  
Differentially Expressed ◽  
F1 Hybrids ◽  
Rna Seq ◽  
Seed Formation ◽  
Maternal Parent ◽  
Expression Of Genes ◽  
Fitness Consequences ◽  
Hybrid Seeds ◽  
Rhinanthus Minor

Germination is a vital stage in a plants life cycle, and a different germination behavior of offspring in comparison to their parents can have fitness consequences. In studies on hybridization between Rhinanthus minor and R. major, low germination rates of F1 hybrids with R. major as the maternal parent have often been reported. In contrast, the F1m hybrid, with R. minor as the maternal parent, germinates readily and rapidly. In order to find the cause of this difference, we used RNA-Seq to obtain transcriptome profiles of F1a and F1m seeds during stratification at 4C and just after germination, after 40 days of stratification for the F1m seeds and 60 days for the F1a seeds. A comparison of the transcriptome of F1a seeds that had just germinated (60 days) with non-germinated F1a seeds after 40 and 60 days revealed 2918 and 1349 differentially expressed (DE) genes, respectively. For F1m seeds, 958 genes showed differential expression in germinated and non-germinated seeds after 40 days. The DE genes of F1a and F1m hybrids clustered into two separate groups, even though they had the same parents, and no differentially expression was found for plastid genes. Non-germinated F1a seeds had an abundance of enzymes and proteins associated with peroxidase activity, peroxiredoxin activity and nutrient reservoir activity. Expression of genes related to seed germination and seed development increased in non-germinated F1a hybrid seeds between 40 and 60 days of cold stratification. F1a seeds that had germinated showed an upregulation of genes related to the gibberellic acid-mediated signaling pathway and response to gibberellin, along with a low expression of DELLA superfamily. Although the results demonstrated strong differences in gene expression during stratification between the reciprocal hybrids, we could not identify its cause, since no plastid genes were differentially expressed. It is possible that differences in embryo development after seed formation and before stratification play a role, including epigenetic imprinting.

scholarly journals LncRNAs Function as Critical Metabolic Regulators in yak Liver by Integrative Transcriptome Analysis

2020 ◽  
Author(s):  
Wei Xia ◽  
Fang Fu ◽  
Li Wang ◽  
Xiaolin Luo ◽  
Jiuqiang Guan
Keyword(s):  
Tibetan Plateau ◽  
Correlation Analysis ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Qrt Pcr ◽  
The People ◽  
Plateau Area ◽  
Metabolic Regulators

Abstract Background: The yak (Bos grunniens) is a crucial resource to supply meat and milk to the people localized in Qinghai-Tibetan plateau area. To identify lncRNAs regulating metabolism in yak, this work adopted transcriptome method to simultaneously profile mRNAs and lncRNAs of liver in yak under three representative age (LD: Liver 1 Day, LM: Liver 15 Months, LY: Liver 5 Years) conditions.Result: Of 288 differentially expressed lncRNAs, function-oriented selection yield 88 regulated metabolically related lncRNAs that were differentially expressed at least two age conditions. These lncRNAs predicted by lncRNA-mRNA correlation analysis to function in various aspects of metabolism. Selected regulations of liver metabolically related lncRNAs were further verified by qRT-PCR.Conclusion: Combining high throughput RNA-seq screening screens, bioinformatics predictions, lncRNA-mRNA correlation analysis and qRT-PCR analysis, this study supports that a class of lncRNAs function as important metabolic regulators and establishes a foundation for further investigating the role of lncRNAs in yak.

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