Expression patterns of MIH, EcR2, and RXR3 in the moult cycle of the red swamp crayfish, Procambarus clarkii (Girard, 1852) (Decapoda, Cambaridae)

Crustaceana ◽  
2019 ◽  
Vol 92 (9) ◽  
pp. 1113-1139
Author(s):  
J. B. Li ◽  
J. G. Ding ◽  
W. Xie ◽  
Y. B. Lin ◽  
H. Li ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Procambarus Clarkii ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Moult Cycle ◽  
Double Stranded Rna ◽  
Eyestalk Ablation ◽  
Red Swamp Crayfish ◽  
Polymerase Chain ◽  
Specific Mrna

Abstract In arthropods, the moult-inhibiting hormone (MIH), the ecdysone receptor (EcR), and the retinoid X receptor (RXR) are key regulators in moulting. In the present study, the full-length cDNAs of the MIH, EcR2, and RXR3 genes from the red swamp crayfish, Procambarus clarkii (Girard, 1852) (denoted as PcMIH, PcEcR2, and PcRXR3) were cloned. Tissue-specific and moult stage-specific mRNA expression patterns of these genes were detected by real-time quantitative polymerase chain reaction. PcMIH was detected only in the eyestalk, whereas PcEcR2 and PcRXR3 mRNA were expressed in all tissues tested. The highest levels of PcEcR2 and PcRXR3 were detected in the gill and hepatopancreas. Expression of PcMIH mRNA in the eyestalk increased from postmoult to peak in intermoult and then decreased in premoult. Expression of PcEcR2 mRNA in the eyestalk, hepatopancreas, and muscle increased from postmoult to peak in early premoult and then decreased. However, expression of PcEcR2 mRNA in the gill increased from postmoult to reach a maximum in intermoult and then decreased in premoult. Expression of PcRXR3 mRNA also fluctuated in the eyestalk, hepatopancreas, muscle, and gill, with a decrease from postmoult to late premoult. Expression of PcEcR2 and PcRXR3 mRNA increased relative to the control in the hepatopancreas and gill after unilateral and bilateral eyestalk ablation, which suggested that PcMIH can inhibit their mRNA expression. Double-stranded RNA-mediated RNA interference of PcRXR3 caused different changes in mRNA expression of these genes in different tissues and resulted in decreased expression of PcEcR2 mRNA, which suggested a collaborative relationship between PcEcR2 and PcRXR3.

scholarly journals Aberrant Whole Blood Gene Expression in the Lumen of Human Intracranial Aneurysms

Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1442
Author(s):  
Vincent M. Tutino ◽  
Yongjun Lu ◽  
Daizo Ishii ◽  
Kerry E. Poppenberg ◽  
Hamidreza Rajabzadeh-Oghaz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Whole Blood ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Transcriptome Profiling ◽  
Circulating Biomarkers ◽  
Post Contrast ◽  
Blood Gene Expression ◽  
Transcriptional Changes ◽  
Polymerase Chain

The rupture of an intracranial aneurysm (IA) causes devastating hemorrhagic strokes. Yet, most IAs remain asymptomatic and undetected until they rupture. In the search for circulating biomarkers of unruptured IAs, we previously performed transcriptome profiling on whole blood and identified an IA-associated panel of 18 genes. In this study, we seek to determine if these genes are also differentially expressed within the IA lumen, which could provide a mechanistic link between the disease and the observed circulating gene expression patterns. To this end, we collected blood from the lumen of 37 IAs and their proximal parent vessels in 31 patients. The expression levels of 18 genes in the lumen and proximal vessel were then measured by quantitative polymerase chain reaction. This analysis revealed that the expression of 6/18 genes (CBWD6, MT2A, MZT2B, PIM3, SLC37A3, and TNFRSF4) was significantly higher in intraluminal blood, while the expression of 3/18 genes (ST6GALNAC1, TCN2, and UFSP1) was significantly lower. There was a significant, positive correlation between intraluminal and proximal expression of CXCL10, MT2A, and MZT2B, suggesting local increases of these genes is reflected in the periphery. Expression of ST6GALNAC1 and TIFAB was significantly positively correlated with IA size, while expression of CCDC85B was significantly positively correlated with IA enhancement on post-contrast MRI, a metric of IA instability and risk. In conclusion, intraluminal expression differences in half of the IA-associated genes observed in this study provide evidence for IA tissue-mediated transcriptional changes in whole blood. Additionally, some genes may be informative in assessing IA risk, as their intraluminal expression was correlated to IA size and aneurysmal wall enhancement.

Wnt/β-Catenin and Hippo Pathway Deregulation in Mammary Tumors of Humans, Dogs, and Cats

2020 ◽  
Vol 57 (6) ◽  
pp. 774-790
Author(s):  
Alessandro Sammarco ◽  
Chiara Gomiero ◽  
Roberta Sacchetto ◽  
Giorgia Beffagna ◽  
Silvia Michieletto ◽  
...  
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Mrna Level ◽  
Hippo Pathway ◽  
Mammary Tumors ◽  
Breast Cancers ◽  
Tumor Tissues ◽  
Mrna And Protein Expression ◽  
Polymerase Chain

Mammary cancer is a common neoplasm in women, dogs, and cats that still represents a therapeutic challenge. Wnt/β-catenin and Hippo pathways are involved in tumor progression, cell differentiation, and metastasis. The aim of this study was to evaluate mRNA and protein expression of molecules involved in these pathways in human (HBC), canine (CMT), and feline mammary tumors (FMT). Real-time quantitative polymerase chain reaction (qPCR) for β-catenin, CCND1, YAP, TAZ, CTGF, and ANKRD1, western blotting for YAP, TAZ, and β-catenin, and immunohistochemistry for estrogen receptor (ER), progesterone receptor (PR), ERBB2, β-catenin, and YAP/TAZ were performed on mammary tumor tissues. The protein expression of active β-catenin was higher in tumors than in healthy tissues in all 3 species. The mRNA expression of the downstream gene CCND1 was increased in HBC ER+ and CMTs compared to healthy tissues. Membranous and cytoplasmic protein expression of β-catenin were strongly negatively correlated in all 3 species. Tumors showed an increased protein expression of YAP/TAZ when compared to healthy tissues. Notably, YAP/TAZ expression was higher in triple negative breast cancers when compared to HBC ER+ and in FMTs when compared to CMTs. The mRNA expression of β-catenin, YAP, TAZ, CTGF, and ANKRD1 was not different between tumors and healthy mammary gland in the 3 species. This study demonstrates deregulation of Wnt/β-catenin and Hippo pathways in mammary tumors, which was more evident at the protein rather than the mRNA level. Wnt/β-catenin and Hippo pathways seem to be involved in mammary carcinogenesis and therefore represent interesting therapeutic targets that should be further investigated.

Gene and protein expression of connexins 37 and 43 in cumulus–oocytes complexes throughout the canine oestrous cycle

2020 ◽  
Vol 32 (11) ◽  
pp. 976
Author(s):  
Monica De los Reyes ◽  
Jaime Palomino ◽  
Carola Gallegos ◽  
Roberto Espinoza ◽  
Phillipe Dettleff ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Cumulus Cells ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Antral Follicles ◽  
Gene And Protein Expression ◽  
Before And After ◽  
Polymerase Chain

The aim of this study was to evaluate the expression of connexin (Cx) 37 and Cx43 in canine cumulus–oocyte complexes (COCs) during the oestrous cycle. Cx localisation was analysed by immunohistochemistry and immunofluorescence, whereas protein and gene expression was evaluated by western blotting and quantitative polymerase chain reaction respectively; comparisons were made using analysis of variance. Both Cx37 and Cx43 were expressed in all follicular stages; Cx43 was identified in cumulus cells and Cx37 was identified in cumulus cells, zonae pellucida and oocytes. Immunofluorescence analyses showed that Cx37 remained unchanged during the preovulatory stage but decreased after ovulation, whereas Cx43 remained unchanged before and after ovulation. Cx43 transcripts increased (P<0.05) during anoestrus and dioestrus in medium-sized follicles but remained unaltered during the pro-oestrus and antral stages during oestrus, before and after ovulation. Cx37 mRNA levels decreased in ovulated COCs (P<0.05). The highest levels of Cx37 protein (P<0.05) were detected in the preantral stage during anoestrus. In contrast, strong Cx43 signals were detected in oestrus and in medium-sized antral follicles in dioestrus (P<0.05). Overall, we demonstrated that Cx37 and Cx43 exhibit different expression patterns, suggesting specific roles throughout growth. Maintenance of Cx expression before ovulation indicates the involvement of Cx37 and Cx43 in the prolonged meiotic arrest.

scholarly journals Ultradian glucocorticoid exposure directs gene-dependent and tissue-specific mRNA expression patterns in vivo

2017 ◽  
Vol 439 ◽  
pp. 46-53 ◽  
Author(s):  
Charlotte L. George ◽  
Matthew T. Birnie ◽  
Benjamin P. Flynn ◽  
Yvonne M. Kershaw ◽  
Stafford L. Lightman ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Expression Patterns ◽  
Tissue Specific ◽  
Specific Mrna

scholarly journals Downregulating a Type I Metacaspase in Petunia Accelerates Flower Senescence

2017 ◽  
Vol 142 (5) ◽  
pp. 405-414 ◽  
Author(s):  
Laura J. Chapin ◽  
Youyoun Moon ◽  
Michelle L. Jones
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Critical Role ◽  
Structural Similarity ◽  
Cysteine Proteases ◽  
Flower Senescence ◽  
Type I ◽  
Environmental Signals ◽  
Petal Development ◽  
Polymerase Chain

Metacaspases are cysteine proteases from plants, fungi, and protozoans that have structural similarity to metazoan caspases. They play a critical role in programmed cell death (PCD) induced by developmental cues and environmental signals. In this study, a type I metacaspase (PhMC1) was identified and characterized from Petunia ×hybrida ‘Mitchell Diploid’ (MD) (petunia). The recombinant PhMC1 had activity against the metacaspase substrate Boc-GRR-AMC (GRR). Activity was highest at pH 7–9 and was dependent on the active site C237. Quantitative polymerase chain reaction (qPCR) showed that PhMC1 transcripts increased at a later stage of petal development, when corollas were visibly senescent in both pollinated and unpollinated flowers. Gene expression patterns were similar to that of the senescence-related gene PhCP10, a homolog of Arabidopsis thaliana (arabidopsis) AtSAG12. PhMC1 transcripts were upregulated in the petals by ethylene treatment. This ethylene regulation did not require protein synthesis, indicating that PhMC1 is a primary ethylene response gene. Metacaspase-like activity against Boc-GRR-AMC increased in protein extracts from senescing petals. RNAi was used to knock down the expression of PhMC1. Transgenic PhMC1 petunias had no abnormal, vegetative growth phenotypes under normal greenhouse conditions, but flower senescence was accelerated by an average of 2 days.

scholarly journals Identification of 118 Arabidopsis Transcription Factor and 30 Ubiquitin-Ligase Genes Responding to Chitin, a Plant-Defense Elicitor

2007 ◽  
Vol 20 (8) ◽  
pp. 900-911 ◽  
Author(s):  
Marc Libault ◽  
Jinrong Wan ◽  
Tomasz Czechowski ◽  
Michael Udvardi ◽  
Gary Stacey
Keyword(s):  
Transcription Factor ◽  
Plant Defense ◽  
Ubiquitin Ligase ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Defense Responses ◽  
Plant Defense Responses ◽  
Genome Array ◽  
Polymerase Chain ◽  
Specific Regulation

Chitin, found in the cell walls of true fungi and the exoskeleton of insects and nematodes, is a well-established elicitor of plant defense responses. In this study, we analyzed the expression patterns of Arabidopsis thaliana transcription factor (TF) and ubiquitin-ligase genes in response to purified chitooctaose at different treatment times (15, 30, 60, 90, and 120 min after treatment), using both quantitative polymerase chain reaction and the Affymetrix Arabidopsis whole-genome array. A total of 118 TF genes and 30 ubiquitin-ligase genes were responsive to the chitin treatment. Among these genes, members from the following four TF families were overrepresented: APETALA2/ethylene-reponsive element binding proteins (27), C2H2 zinc finger proteins (14), MYB domain-containing proteins (11), and WRKY domain transcription factors (14). Transcript variants from a few of these genes were found to respond differentially to chitin, suggesting transcript-specific regulation of these TF genes.

scholarly journals Identification and Characterization of Osmoregulation Related MicroRNAs in Gills of Hybrid Tilapia Under Three Types of Osmotic Stress

2021 ◽  
Vol 12 ◽  
Author(s):  
Huanhuan Su ◽  
Jiajia Fan ◽  
Dongmei Ma ◽  
Huaping Zhu
Keyword(s):  
Polymerase Chain Reaction ◽  
Osmotic Stress ◽  
Expression Profiles ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
Protein Translation ◽  
Control Group ◽  
Alkali Stress ◽  
Chain Reaction ◽  
Polymerase Chain

Researchers have increasingly suggested that microRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression and protein translation in organs and respond to abiotic and biotic stressors. To understand the function of miRNAs in osmotic stress regulation of the gills of hybrid tilapia (Oreochromis mossambicus ♀ × Oreochromis urolepis hornorum ♂), high-throughput Illumina deep sequencing technology was used to investigate the expression profiles of miRNAs under salinity stress (S, 25‰), alkalinity stress (A, 4‰) and salinity–alkalinity stress (SA, S: 15‰, A: 4‰) challenges. The results showed that 31, 41, and 27 upregulated and 33, 42, and 40 downregulated miRNAs (P < 0.05) were identified in the salt stress, alkali stress, and saline–alkali stress group, respectively, which were compared with those in the control group (C). Fourteen significantly differently expressed miRNAs were selected randomly and then validated by a quantitative polymerase chain reaction. On the basis of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, genes related to osmoregulation and biosynthesis were enriched in the three types of osmotic stress. In addition, three miRNAs and three predicted target genes were chosen to conduct a quantitative polymerase chain reaction in the hybrid tilapia and its parents during 96-h osmotic stress. Differential expression patterns of miRNAs provided the basis for research data to further investigate the miRNA-modulating networks in osmoregulation of teleost.

scholarly journals Biosensor of inflammation biomarkers based on electrical bioimpedance analysis on immobilized DNA without chemical modification

2020 ◽  
Vol 11 (1) ◽  
pp. 31-37
Author(s):  
Modesto Gómez-López ◽  
Ángel Miliar-García ◽  
Nadia Mabel Pérez-Vielma ◽  
Eleazar Lara-Padilla ◽  
César Antonio González-Díaz
Keyword(s):  
Gene Expression ◽  
Dna Sequences ◽  
Deoxyribonucleic Acid ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Patterns ◽  
New Techniques ◽  
Detection Techniques ◽  
Electrical Bioimpedance ◽  
Polymerase Chain ◽  
Inexpensive Technique

AbstractThe development of biosensors to identify molecular markers or specific genes is fundamental for the implementation of new techniques that allow the detection of specific Deoxyribonucleic acid (DNA) sequences in a fast, economic and simple way. Different detection techniques have been proposed in the development of biosensors. Electrical Bioimpedance Spectroscopy (EBiS) has been used for diagnosis and monitoring of human pathologies, and is recognized as a safe, fast, reusable, easy and inexpensive technique. This study proves the development of a complementary DNA (cDNA) biosensor based on measurements of EBiS and DNA's immobilization with no chemical modifications. The evaluation of its potential utility in the detection of the gene expression of three inflammation characteristic biomarkers (NLRP3, IL-1β and Caspase 1) is presented. The obtained results demonstrate that EBiS can be used to identify different gene expression patterns, measurements that were validated by Quantitative Polymerase Chain Reaction (qPCR). These results indicate the technical feasibility for a biosensor of specific genes through bioimpedance measurements on the immobilization of cDNA.

scholarly journals Expression of the Multidrug Resistance-Associated Protein Gene in Refractory Lymphoma: Quantitation by a Validated Polymerase Chain Reaction Assay

Blood ◽  
1997 ◽  
Vol 89 (10) ◽  
pp. 3795-3800 ◽  
Author(s):  
Zhirong Zhan ◽  
Victor A. Sandor ◽  
Erick Gamelin ◽  
Joanna Regis ◽  
Bruce Dickstein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multidrug Resistance ◽  
Mrna Expression ◽  
Quantitative Polymerase Chain Reaction ◽  
Newly Diagnosed ◽  
Chain Reaction ◽  
Significant Difference ◽  
Polymerase Chain ◽  
Refractory Lymphoma ◽  
Mdr 1

Abstract Previous work investigating the role of MDR-1 overexpression in relapsed and refractory lymphoma led us to investigate a possible role for multidrug resistance-associated protein (MRP) as a cause of resistance in patients who did not overexpress MDR-1. A quantitative polymerase chain reaction (PCR) method for measuring MRP expression was validated. Immunoblot analysis suggested that no major discrepancy was present between mRNA expression and protein levels. MRP levels were found to be independent of sample tumor content by immunophenotyping, suggesting that the presence of normal cells had no significant impact on measurements of MRP expression. We evaluated MRP in 55 biopsy samples from 40 patients with refractory lymphoma enrolled on a trial of infusional chemotherapy (EPOCH). Pre- and post-EPOCH samples were available from 15 patients. MRP levels were also evaluated in 16 newly diagnosed, untreated lymphoma patient samples. No significant difference in MRP mRNA expression was noted between pre- and post-EPOCH groups. Also, MRP levels in the newly diagnosed patient samples were not significantly different from either pre- or post-EPOCH groups. Two of 15 paired pre- and post-EPOCH patient samples exhibited overexpression of MRP after EPOCH chemotherapy, with measured increases of 10-fold and 18-fold. We conclude that MRP overexpression is not responsible for non–P-glycoprotein (Pgp)–mediated drug resistance in the majority of these patients, although it may be important in a subset of patients. Defining this subset prospectively could aid in the development of clinical trials of MRP modulation in drug-resistant lymphoma.

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