scholarly journals Effects of Species Differences on Oocyte Regulation of Granulosa Cell Proliferation

2021 ◽  
Author(s):  
◽  
Jia Yi Lin
Keyword(s):  
Gene Expression ◽  
Growth Factors ◽  
Incubation Period ◽  
Species Differences ◽  
Thymidine Uptake ◽  
Mrna Levels ◽  
Expression Levels ◽  
Spent Media ◽  
Over Time ◽  
Stimulation Of

<p>The role of oocytes in regulating ovulation quota between species is not fully understood. In humans, sheep, and rodents the oocyte-derived growth factors, bone morphogenetic protein 15 (BMP15) and/or growth differentiation factor 9 (GDF9) have profound effects on ovarian follicular development and ovulation quota. The aim of these studies was to compare the ability of oocytes from sheep (low ovulation-rate) and rat (poly-ovulator) to stimulate radiolabelled ³H-thymidine uptake by granulosa cells (GC) both within and between the two species, and to assess species differences of GDF9 and BMP15 in co-incubations. For these experiments, oocytes denuded of cumulus-cells (DO) were co-incubated with a fixed number of GC from either species. Rat or sheep DO stimulated ³H-thymidine uptake by GC from the same species (P<0.005). Sheep oocytes also stimulated ³H-thymidine uptake by rat GC (P<0.001) but not vice versa. To investigate this further, oocytes and GC were co-incubated with monoclonal antibodies specific to GDF9 or BMP15 or to a hydatids antigen (control). Both sheep and rat oocyte stimulation of ³H-thymidine uptake by GC was inhibited with the GDF9 antibody (P<0.05) but not the control antibody, irrespective of the species of GC. Sheep DO stimulation of rat GC was also inhibited using an antibody to BMP15 (P<0.05). However, when using the BMP15 antibody to block the effects of rat DO on rat GC, no inhibition of ³H-thymidine uptake was observed (P=0.988). The molecular forms of GDF9 and BMP15 protein in oocyte lysates and spent media were examined by Western blotting under reducing conditions. For both species, GDF9 protein was present in the mature form in both the lysate and the spend media. For sheep oocytes, BMP15 protein was present as pro-mature and monomeric mature forms in the lysate and in the spent media, whereas from rats pro-mature forms were detected in the oocyte lysate but there was minimal evidence for the mature form in the spent media. The mRNA levels of GDF9, BMP15 and several cumulus cell (CC) genes were measured at 8h intervals over a 24h incubation period. For both sheep and the rat, GDF9 and BMP15 mRNA expression levels were highly correlated (R²=0.99). In the rat, the relative mean expression level of Gdf9 mRNA was four times higher than Bmp15 mRNA, whereas in sheep the relative mean expression ratio of GDF9 to BMP15 was approximately one. For these studies, two types of incubations were performed, namely COC alone or co-incubations of DO with GC. In co-incubations of DO with GC, the relative GDF9 and BMP15 mRNA levels did not change significantly over time, whereas incubations of COC showed that the relative levels of GDF9 and BMP15 mRNA declined significantly during the incubation period. The CC genes (FSHR, LHR, KITL, CX43, AROM and CYCD2) were measured during the 24h incubation period for the COC but not the DO experiment. In rats, none of the CC genes showed any changes in mRNA levels over time except Kitl, where levels at 8h and 16h mRNA levels were significantly lower compared to those at 0h and 24h. In sheep, there were no significant mRNA changes in any of the CC genes except AROM. The level of AROM mRNA reduced to near zero by 8h and remained at low levels at both 16h and 24h. These findings suggest that, under the incubation conditions used, that the gene expression levels for most CC genes were maintained but that the oocyte growth factors alone were unable to maintain AROM gene expression in sheep. In conclusion, major differences were observed in GDF9 and BMP15 expression and regulation between rats and sheep. The lower expression levels of Bmp15 mRNA and protein are likely the reason why rat DO failed to stimulate ³H-thymidine uptake by sheep GC. The evidence suggests that oocyte-derived GDF9 is sufficient to stimulate ³H-thymidine incorporation and presumably DNA synthesis in rat GC whereas in sheep both BMP15 and GDF9 are required. These findings raise the possibility that in species with a low ovulation rate phenotype there is a higher level of GDF9 mRNA and protein synthesis and that BMP15 is a major factor in restricting ovulation quota in mammals.</p>

scholarly journals Effects of Species Differences on Oocyte Regulation of Granulosa Cell Proliferation

2021 ◽  
Author(s):  
◽  
Jia Yi Lin
Keyword(s):  
Gene Expression ◽  
Growth Factors ◽  
Incubation Period ◽  
Species Differences ◽  
Thymidine Uptake ◽  
Mrna Levels ◽  
Expression Levels ◽  
Spent Media ◽  
Over Time ◽  
Stimulation Of

<p>The role of oocytes in regulating ovulation quota between species is not fully understood. In humans, sheep, and rodents the oocyte-derived growth factors, bone morphogenetic protein 15 (BMP15) and/or growth differentiation factor 9 (GDF9) have profound effects on ovarian follicular development and ovulation quota. The aim of these studies was to compare the ability of oocytes from sheep (low ovulation-rate) and rat (poly-ovulator) to stimulate radiolabelled ³H-thymidine uptake by granulosa cells (GC) both within and between the two species, and to assess species differences of GDF9 and BMP15 in co-incubations. For these experiments, oocytes denuded of cumulus-cells (DO) were co-incubated with a fixed number of GC from either species. Rat or sheep DO stimulated ³H-thymidine uptake by GC from the same species (P<0.005). Sheep oocytes also stimulated ³H-thymidine uptake by rat GC (P<0.001) but not vice versa. To investigate this further, oocytes and GC were co-incubated with monoclonal antibodies specific to GDF9 or BMP15 or to a hydatids antigen (control). Both sheep and rat oocyte stimulation of ³H-thymidine uptake by GC was inhibited with the GDF9 antibody (P<0.05) but not the control antibody, irrespective of the species of GC. Sheep DO stimulation of rat GC was also inhibited using an antibody to BMP15 (P<0.05). However, when using the BMP15 antibody to block the effects of rat DO on rat GC, no inhibition of ³H-thymidine uptake was observed (P=0.988). The molecular forms of GDF9 and BMP15 protein in oocyte lysates and spent media were examined by Western blotting under reducing conditions. For both species, GDF9 protein was present in the mature form in both the lysate and the spend media. For sheep oocytes, BMP15 protein was present as pro-mature and monomeric mature forms in the lysate and in the spent media, whereas from rats pro-mature forms were detected in the oocyte lysate but there was minimal evidence for the mature form in the spent media. The mRNA levels of GDF9, BMP15 and several cumulus cell (CC) genes were measured at 8h intervals over a 24h incubation period. For both sheep and the rat, GDF9 and BMP15 mRNA expression levels were highly correlated (R²=0.99). In the rat, the relative mean expression level of Gdf9 mRNA was four times higher than Bmp15 mRNA, whereas in sheep the relative mean expression ratio of GDF9 to BMP15 was approximately one. For these studies, two types of incubations were performed, namely COC alone or co-incubations of DO with GC. In co-incubations of DO with GC, the relative GDF9 and BMP15 mRNA levels did not change significantly over time, whereas incubations of COC showed that the relative levels of GDF9 and BMP15 mRNA declined significantly during the incubation period. The CC genes (FSHR, LHR, KITL, CX43, AROM and CYCD2) were measured during the 24h incubation period for the COC but not the DO experiment. In rats, none of the CC genes showed any changes in mRNA levels over time except Kitl, where levels at 8h and 16h mRNA levels were significantly lower compared to those at 0h and 24h. In sheep, there were no significant mRNA changes in any of the CC genes except AROM. The level of AROM mRNA reduced to near zero by 8h and remained at low levels at both 16h and 24h. These findings suggest that, under the incubation conditions used, that the gene expression levels for most CC genes were maintained but that the oocyte growth factors alone were unable to maintain AROM gene expression in sheep. In conclusion, major differences were observed in GDF9 and BMP15 expression and regulation between rats and sheep. The lower expression levels of Bmp15 mRNA and protein are likely the reason why rat DO failed to stimulate ³H-thymidine uptake by sheep GC. The evidence suggests that oocyte-derived GDF9 is sufficient to stimulate ³H-thymidine incorporation and presumably DNA synthesis in rat GC whereas in sheep both BMP15 and GDF9 are required. These findings raise the possibility that in species with a low ovulation rate phenotype there is a higher level of GDF9 mRNA and protein synthesis and that BMP15 is a major factor in restricting ovulation quota in mammals.</p>

scholarly journals Methylobacterium oryzae Influences Isoepoxydon Dehydrogenase Gene Expression and Patulin Production by Penicillium expansum

Water ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1427
Author(s):  
Tiago Barros Afonso ◽  
Lúcia Chaves Simões ◽  
Nelson Lima
Keyword(s):  
Gene Expression ◽  
Distribution Systems ◽  
Water Distribution ◽  
Penicillium Expansum ◽  
Transcriptional Level ◽  
Positive Trend ◽  
Mrna Levels ◽  
Expression Levels ◽  
Production Values ◽  
Methylobacterium Oryzae

Biofilms can be considered the main source of microorganisms in drinking water distribution systems (DWDS). The ecology of a biofilm is dependent on a variety of factors, including the presence of microbial metabolites excreted by its inhabitants. This study reports the effect of the Gram-negative bacteria Methylobacterium oryzae on the idh gene expression levels and patulin production of Penicillium expansum mature biofilms. For this purpose, a RT-qPCR method to quantify idh mRNA levels was applied. In addition, the idh expression levels were compared with the patulin production. The results obtained revealed that the effect of the bacterium on pre-established P. expansum biofilms is dependent on the time of interaction. More mature P. expansum biofilms appear to be more resistant to the inhibitory effect that M. oryzae causes towards idh gene expression and patulin production. A positive trend was observed between the idh expression and patulin production values. The results indicate that M. oryzae affects patulin production by acting at the transcriptional level of the idh gene.

scholarly journals Gene Expression of Nucleic Acid-Sensing Pattern Recognition Receptors in Children Hospitalized for Respiratory Syncytial Virus-Associated Acute Bronchiolitis

2009 ◽  
Vol 16 (6) ◽  
pp. 816-823 ◽  
Author(s):  
Carolina Scagnolari ◽  
Fabio Midulla ◽  
Alessandra Pierangeli ◽  
Corrado Moretti ◽  
Enea Bonci ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pattern Recognition ◽  
Viral Load ◽  
Respiratory Syncytial Virus ◽  
Pattern Recognition Receptors ◽  
Mrna Levels ◽  
Acute Bronchiolitis ◽  
Expression Levels ◽  
Rsv Infection ◽  
Syncytial Virus

ABSTRACT Given the critical role of pattern recognition receptors (PRRs) in acid nucleic recognition in the initiation of innate immunity and the orchestration of adaptive immunity, the aim of this study was to determine whether any heterogeneity of PRR expression in the airway tracts of infants with respiratory syncytial virus (RSV) infection might explain the broad clinical spectrum of RSV-associated bronchiolitis in infants. For this purpose, the levels of melanoma differentiation-associated protein-5 (MDA-5), retinoic acid inducible gene-1 (RIG-1), and Toll-like receptor 3 (TLR-3), TLR-7, TLR-8, and TLR-9 mRNAs were evaluated, using TaqMan quantitative reverse transcription-PCR, in cells from nasopharyngeal washes collected from 157 infants suffering from acute bronchiolitis whether or not they were associated with respiratory viruses. High interindividual variability was observed in both virus-positive and -negative infants; however, the relative gene expression levels of MDA-5, RIG-1, TLR-7, and TLR-8 were significantly higher in the virus-infected group, whereas the expression levels of TLR-3 and TLR-9 were not significantly different. The differences in the gene expression of MDA-5, RIG-1, TLR-7, and TLR-8 were more evident in infants with RSV infection than in those with bocavirus or rhinovirus infection. In RSV-infected infants, PRR-mRNA levels also were analyzed in relation to interferon protein levels, viral load, clinical severity, days of hospitalization, age, and body weight. A significant positive correlation was observed only between RSV viral load and RIG-1 mRNA levels. These findings provide the first direct evidence that, in infants with respiratory virus-associated bronchiolitis, especially RSV, there are substantial changes in PRR gene expression; this likely is an important determinant of the clinical outcome of bronchiolitis.

scholarly journals Jonckheere–Terpstra–Kendall-based non-parametric analysis of temporal differential gene expression

2021 ◽  
Vol 3 (1) ◽  
Author(s):  
Hitoshi Iuchi ◽  
Michiaki Hamada
Keyword(s):  
Gene Expression ◽  
Time Series ◽  
Time Course ◽  
Time Series Data ◽  
Expression Patterns ◽  
Detection Methods ◽  
Series Data ◽  
Expression Levels ◽  
Over Time ◽  
Non Parametric

Abstract Time-course experiments using parallel sequencers have the potential to uncover gradual changes in cells over time that cannot be observed in a two-point comparison. An essential step in time-series data analysis is the identification of temporal differentially expressed genes (TEGs) under two conditions (e.g. control versus case). Model-based approaches, which are typical TEG detection methods, often set one parameter (e.g. degree or degree of freedom) for one dataset. This approach risks modeling of linearly increasing genes with higher-order functions, or fitting of cyclic gene expression with linear functions, thereby leading to false positives/negatives. Here, we present a Jonckheere–Terpstra–Kendall (JTK)-based non-parametric algorithm for TEG detection. Benchmarks, using simulation data, show that the JTK-based approach outperforms existing methods, especially in long time-series experiments. Additionally, application of JTK in the analysis of time-series RNA-seq data from seven tissue types, across developmental stages in mouse and rat, suggested that the wave pattern contributes to the TEG identification of JTK, not the difference in expression levels. This result suggests that JTK is a suitable algorithm when focusing on expression patterns over time rather than expression levels, such as comparisons between different species. These results show that JTK is an excellent candidate for TEG detection.

scholarly journals Heterogeneous recruitment abilities to RNA polymerases generate nonlinear scaling of gene expression with cell volume

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Qirun Wang ◽  
Jie Lin
Keyword(s):  
Gene Expression ◽  
Cell Volume ◽  
Direct Consequence ◽  
Rna Polymerases ◽  
Mrna Levels ◽  
Expression Levels ◽  
Promoter Sequences ◽  
Biophysical Mechanism ◽  
Expression Model ◽  
Cycle Regulation

AbstractWhile most genes’ expression levels are proportional to cell volumes, some genes exhibit nonlinear scaling between their expression levels and cell volume. Therefore, their mRNA and protein concentrations change as the cell volume increases, which often have crucial biological functions such as cell-cycle regulation. However, the biophysical mechanism underlying the nonlinear scaling between gene expression and cell volume is still unclear. In this work, we show that the nonlinear scaling is a direct consequence of the heterogeneous recruitment abilities of promoters to RNA polymerases based on a gene expression model at the whole-cell level. Those genes with weaker (stronger) recruitment abilities than the average ability spontaneously exhibit superlinear (sublinear) scaling with cell volume. Analysis of the promoter sequences and the nonlinear scaling of Saccharomyces cerevisiae’s mRNA levels shows that motifs associated with transcription regulation are indeed enriched in genes exhibiting nonlinear scaling, in concert with our model.

scholarly journals Codon usage is an important determinant of gene expression levels largely through its effects on transcription

2016 ◽  
Vol 113 (41) ◽  
pp. E6117-E6125 ◽  
Author(s):  
Zhipeng Zhou ◽  
Yunkun Dang ◽  
Mian Zhou ◽  
Lin Li ◽  
Chien-hung Yu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Codon Usage ◽  
Important Determinant ◽  
Mrna Translation ◽  
Mrna Levels ◽  
Protein Coding ◽  
Expression Levels ◽  
Highly Expressed Genes ◽  
The Impact ◽  
Gene Expression Levels

Codon usage biases are found in all eukaryotic and prokaryotic genomes, and preferred codons are more frequently used in highly expressed genes. The effects of codon usage on gene expression were previously thought to be mainly mediated by its impacts on translation. Here, we show that codon usage strongly correlates with both protein and mRNA levels genome-wide in the filamentous fungus Neurospora. Gene codon optimization also results in strong up-regulation of protein and RNA levels, suggesting that codon usage is an important determinant of gene expression. Surprisingly, we found that the impact of codon usage on gene expression results mainly from effects on transcription and is largely independent of mRNA translation and mRNA stability. Furthermore, we show that histone H3 lysine 9 trimethylation is one of the mechanisms responsible for the codon usage-mediated transcriptional silencing of some genes with nonoptimal codons. Together, these results uncovered an unexpected important role of codon usage in ORF sequences in determining transcription levels and suggest that codon biases are an adaptation of protein coding sequences to both transcription and translation machineries. Therefore, synonymous codons not only specify protein sequences and translation dynamics, but also help determine gene expression levels.

scholarly journals Induction of c-ets and c-fos gene expression upon antigenic stimulation of a T cell hybridoma with inducible cytolytic capacity.

1987 ◽  
Vol 166 (3) ◽  
pp. 810-815 ◽  
Author(s):  
Y Kaufmann ◽  
T Silverman ◽  
B Z Levi ◽  
K Ozato
Keyword(s):  
Gene Expression ◽  
T Cell ◽  
T Cell Activation ◽  
Cell Activation ◽  
Specific Antigen ◽  
Functional Differentiation ◽  
Mrna Levels ◽  
Cellular Oncogenes ◽  
Differentiation Of Lymphocytes ◽  
Stimulation Of

Expression of cellular oncogenes was studied in a T cell hybridoma that undergoes cytolytic activation when stimulated by specific antigen or by anti-Thy-1 antibody. The activation occurs without induction of hybridoma proliferation, providing a model to examine oncogene expression during functional differentiation of lymphocytes. We found that c-fos and c-ets-1 mRNAs were transiently induced at high levels in the hybridoma 30 min and 4 h after stimulation, respectively. c-myc and c-ets-2 oncogenes were constitutively expressed in the hybridoma and their mRNA levels were unaffected during 4 h of stimulation, although c-myc expression was reduced in the later stage of stimulation. Inhibitors of T cell activation, cyclosporin A and anti-LFA-1 antibody, blocked the induction of c-fos and c-ets-1 mRNAs without reducing the levels of c-myc and c-ets-2. The results indicate that the functional activation of the CTL hybridoma is associated with induction of c-fos and c-ets-1 genes.

ERCC1 and RRM1 but not EGFR gene expression is predictive of shorter survival in advanced non-small cell lung cancer (NSCLC)

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 10030-10030
Author(s):  
G. Selvaggi ◽  
P. Ceppi ◽  
M. Volante ◽  
S. Saviozzi ◽  
S. Novello ◽  
...  
Keyword(s):  
Gene Expression ◽  
Excision Repair ◽  
Growth Factor Receptor ◽  
Needle Aspiration ◽  
Differential Sensitivity ◽  
Mrna Levels ◽  
Female Ratio ◽  
Expression Levels ◽  
Egfr Expression ◽  
Formalin Fixed Paraffin Embedded

10030 Background: Pivotal studies indicate a role of excision repair cross-complementation 1 (ERCC1) and ribonucleotide reductase M1 (RRM1) in conferring a differential sensitivity to cytotoxic chemotherapy and epidermal growth factor receptor (EGFR) has been recently deeply investigated in NSCLC. Methods: We retrospectively collected 70 formalin-fixed paraffin-embedded (FFPE) bronchoscopic/fine needle aspiration biopsies of NSCLC to investigate the expression levels of ERCC1, RRM1 and EGFR by Real-Time PCR (Lord R et al. Clin Cancer Research 2002, 8:2286–91). Results were correlated with survival using the Kaplan-Meier method. Results: Sixty-one (87%) specimens were successfully amplified. Median age was 62 years (range 26–75), male/ female ratio 44/17, stage III/IV 20/41; 43 patients received cisplatin-based chemotherapy; overall median survival (MS) was 13.3 months over a median follow-up time of 45 months. ERCC1 expression level ranged from 0.70 to 15.12, RRM1 0.60–17.82. By adopting cut-off values according to median expression levels, we found a strong correlation between ERCC1 and RRM1 mRNA levels (r=0.410; p<0.001). MS in patients with low ERCC1 was significantly longer (16.9 vs 11.3 months, p<0.006) as well as in patients with low RRM1 (13.9 vs 10.9 months, p<0.03). Concomitant high expression levels of ERCC1 and RRM1 (n=26) are predictive of a worse outcome (13.9 vs 10.9 months, p<0.05). Among patients treated with cisplatin-based regimens, low ERCC1 levels were also predictive of a significantly longer MS (23.0 vs 11.6 months, p<0.002). A lower median ERCC1 level (3.2 vs 4.7) and a correlation with a better outcome were also observed in females vs males. No correlation between gene expression levels and histology was reported. No significant correlation between EGFR expression levels (range 0.5–85.8) and survival was found, even when different cut-off values were tested. Conclusions: This retrospective study further validates ERCC1 and RRM1 as good candidates genes to customize chemotherapy. Prospective studies based on the selection of patients according to genes expression levels are a research priority in early and advanced stages of NSCLC. [Table: see text]

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