scholarly journals Gonadotropin Stimulation Has Only a Limited Effect on the Concentration of Follicular Fluid Signalling Proteins: An Antibody Array Analysis

2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Nick A. Bersinger ◽  
Markus Eisenhut ◽  
Petra Stute ◽  
Michael von Wolff
Keyword(s):  
Map Kinase ◽  
Follicular Fluid ◽  
Caspase 3 ◽  
Differentially Expressed ◽  
Matched Pair ◽  
In Vitro Fertilisation ◽  
Array Analysis ◽  
Gonadotropin Stimulation ◽  
Cystatin A ◽  
Follicular Fluids

Objective. The follicular fluid (FF) plays an essential role in the physiology of the follicle and the oocyte. Gonadotropin stimulation affects the FF steroid hormone and anti-Mullerian hormone (AMH) concentrations, which has been suggested to be the reason for lower oocyte competence in conventional gonadotropin stimulated in vitro fertilisation (cIVF) compared to natural cycle IVF (NC-IVF). To analyse the effect of gonadotropin stimulation on a broad spectrum of signalling proteins, we ran proteomic antibody arrays on FF of women undergoing both treatments NC-IVF and cIVF. Method. Twenty women underwent one NC-IVF and one cIVF treatment cycle. Follicular fluids of the first aspirated follicle were compared between the two groups using a protein microarray which included antibodies against 224 proteins related to cell signalling and reference proteins. Each of the 40 albumin-stripped, matched-pair samples was labelled in the reverse-dye (Cy3/Cy5) procedure before undergoing array hybridisation. Signal analysis was performed using normalisation algorithms in dedicated software. Five proteins yielding a value of P < 0.05 in the array experiment (Cystatin A, Caspase-3, GAD65/67, ERK-1, and ERK-2) were then submitted to quantitative determination by ELISA in the same follicular fluids. Results. Array analysis yielded only a small number of differentially expressed signalling markers by unadjusted P values. Adjustment as a consequence of multiple determinations resulted in the absence of any significant differential marker expression on the array. Five unadjusted differentially expressed proteins were quantified immunometrically with antibodies from different sources. Follicular fluid concentrations of Cystatin A and MAP kinase ERK-1 concentrations were significantly higher in the cIVF than in the NC-IVF follicles, while GAD-2 (GAD65/67) did not differ. The assays for Caspase-3 and MAP kinase ERK-2 did not have the required sensitivities. Conclusion. In contrast to FF steroid hormones and AMH, FF concentrations of signalling proteins are not or only marginally altered by gonadotropin stimulation.

scholarly journals Significance of disorders of proteomic profile of follicular fluid for predicting the effectiveness of in vitro fertilisation in women with infertility

2018 ◽  
Vol 99 (3) ◽  
pp. 496-503
Author(s):  
O S Zolotykh ◽  
S V Lomteva ◽  
K Yu Sagamonova
Keyword(s):  
Assisted Reproductive Technology ◽  
Follicular Fluid ◽  
Molecular Mechanisms ◽  
Reproductive Technologies ◽  
Reproductive Technology ◽  
In Vitro Fertilisation ◽  
Proteomic Profile ◽  
Technology Programs ◽  
Group 2

Aim. To study the proteomic profile of follicular fluid in patients with infertility in assisted reproductive technology programs. Methods. The study included women with infertility included in assisted reproductive technology programs: 15 women who had in vitro fertilisation which resulted in pregnancy (group 1) and 16 women with a negative result of this program (group 2). Fractionation of the follicular fluid samples was performed using the sets of special magnetic beads. Proteomic profiling was performed by tandem MALDI-mass-spectrometry. The anti-Müllerian hormone level was measured by ELISA. Results. The study revealed differences in the detectability of follicular fluid proteins with different regulatory properties in patients of groups 1 and 2. With the negative outcome of in vitro fertilisation, expression of a number of proteins involved in the processes of folliculogenesis, ovulation, selection of the dominant follicle, as well as proteins necessary for the development of the zygote and blastula was reduced in females' follicular fluid. Increased expression in women from group 2 was registered for proteins enhancing proteolytic reactions, cell apoptosis, including oocytes, which disrupt the positive action of activin and damage structural and functional state of mitochondria. A definite relationship was found between the level of anti-Müllerian hormone and rate of detection of a number of proteins, in particular protocadherin-2α, cystatin C, betaglycan, prostatic acid phosphatase, and dermicidin. Conclusion. The revealed changes in proteomic profile of the follicular fluid obviously play an important role in the molecular mechanisms that determine the effectiveness of assisted reproductive technologies; the identified differentially expressed proteins can serve as objective markers for predicting the outcomes of in vitro fertilisation.

Role of F-actin organization in p38 MAP kinase-mediated apoptosis and necrosis in neonatal rat cardiomyocytes subjected to simulated ischemia and reoxygenation

2005 ◽  
Vol 289 (6) ◽  
pp. H2310-H2318 ◽  
Author(s):  
Takayuki Okada ◽  
Hajime Otani ◽  
Yue Wu ◽  
Shiori Kyoi ◽  
Chiharu Enoki ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Map Kinase ◽  
Dna Fragmentation ◽  
P38 Mapk ◽  
Caspase 3 ◽  
Neonatal Rat ◽  
Cytochrome C Release ◽  
Actin Reorganization ◽  
Ldh Release ◽  
Sb 203580

Activation of p38 mitogen-activated protein (MAP) kinase (MAPK) has been implicated in the mechanism of cardiomyocyte (CMC) protection and injury. The p38 MAPK controversy may be related to differential effects of this kinase on apoptosis and necrosis. We have hypothesized that p38 MAPK-mediated F-actin reorganization promotes apoptotic cell death, whereas it protects from osmotic stress-induced necrotic cell death. Cultured neonatal rat CMCs were subjected to 2 h of simulated ischemia followed by reoxygenation. p38 MAPK activity measured by phosphorylation of MAP kinase-activated protein (MAPKAP) kinase 2 was increased during simulated ischemia and reoxygenation. This was associated with translocation of heat shock protein 27 (HSP27) from the cytosolic to the cytoskeletal fraction and F-actin reorganization. Cytochrome c release from mitochondria, caspase-3 activation, and DNA fragmentation were increased during reoxygenation. Robust lactate dehydrogenase (LDH) release was observed under hyposmotic (140 mosM) reoxygenation. The p38 MAPK inhibitor SB-203580 abrogated activation of p38 MAPK, translocation of HSP27, and F-actin reorganization and prevented cytochrome c release, caspase-3 activation, and DNA fragmentation. Conversely, SB-203580 enhanced LDH release during hyposmotic reoxygenation. The F-actin disrupting agent cytochalasin D inhibited F-actin reorganization and prevented cytochrome c release, caspase-3 activation, and DNA fragmentation, whereas it enhanced LDH release during hyposmotic reoxygenation. When CMCs were incubated under the isosmotic condition for the first 15 min of reoxygenation, SB-203580 and cytochalasin D increased ATP content of CMCs and prevented LDH release after the conversion to the hyposmotic condition. These results suggest that F-actin reorganization mediated by activation of p38 MAPK plays a differential role in apoptosis and protection against osmotic stress-induced necrosis during reoxygenation in neonatal rat CMCs; however, the sarcolemmal fragility caused by p38 MAPK inhibition can be reversed during temporary blockade of physical stress during reoxygenation.

scholarly journals Proteomic analysis of follicular fluid in carriers and non-carriers of the Trio allele for high ovulation rate in cattle

2018 ◽  
Vol 30 (12) ◽  
pp. 1643 ◽  
Author(s):  
Mamat H. Kamalludin ◽  
Alvaro Garcia-Guerra ◽  
Milo C. Wiltbank ◽  
Brian W. Kirkpatrick
Keyword(s):  
Follicular Fluid ◽  
Transforming Growth Factor ◽  
Fluid Collection ◽  
Transforming Growth Factor Β ◽  
Initial Study ◽  
Ovulation Rate ◽  
Differentially Expressed ◽  
Masking Effect ◽  
Joint Analysis ◽  
Abundance Proteins

This study was conducted to characterise differences in follicular fluid proteins between carriers and non-carriers of a bovine allele for high ovulation rate. A total of four non-carrier and five carrier females were used in an initial study with four and six additional non-carriers and carriers respectively used in a validation study. Emergence of the follicular wave was synchronised and the ovaries containing the dominant follicle(s) were extracted by ovariectomy for follicular fluid collection. A hexapeptide ligand library was used to overcome the masking effect of high-abundance proteins and to increase detection of low-abundance proteins in tandem mass spectrometry. After correcting for multiple comparisons, only two proteins, glia-derived nexin precursor (SERPINE2) and inhibin β B chain precursor (INHBB), were significantly differentially expressed (false-discovery rate <0.05). In a replicate study of analogous design differential expression was confirmed (P < 0.05). Joint analysis of results from the two studies indicated that three additional proteins were consistently differentially expressed between genotypes. For three of these five, previous studies have indicated that expression is increased by transforming growth factor-β–bone morphogenetic protein signalling; their reduction in follicular fluid from carrier animals is consistent with the ~9-fold overexpression of SMAD family member 6 (SMAD6) in carriers that is inhibitory to this pathway.

scholarly journals Differential expression of CART in ewes with differing ovulation rates

Reproduction ◽  
2017 ◽  
Vol 153 (4) ◽  
pp. 471-479 ◽  
Author(s):  
Jennifer L Juengel ◽  
Michelle C French ◽  
Laurel D Quirke ◽  
Alexia Kauff ◽  
George W Smith ◽  
...  
Keyword(s):  
Differential Expression ◽  
Expression Pattern ◽  
Follicular Fluid ◽  
Follicular Phase ◽  
Distinct Pattern ◽  
Ovulation Rate ◽  
Differentially Expressed ◽  
Antral Follicles ◽  
Cart Peptide

We hypothesised that cocaine- and amphetamine-regulated transcript (CARTPT) would be differentially expressed in ewes with differing ovulation rates. Expression of mRNA forCARTPT, as well asLHCGR,FSHR,CYP19A1andCYP17A1was determined in antral follicles ≥1 mm in diameter collected during the follicular phase in ewes heterozygous for the Booroola and Inverdale genes (I+B+; average ovulation rate 4) and ++ contemporaries (++; average ovulation rate 1.8). In ++ ewes (n = 6),CARTPTwas expressed in small follicles (1 to <3 mm diameter), where 18.8 ± 2.5% follicles expressedCARTPT. CART peptide was also detected in follicular fluid of some follicles of ++ ewes. In I+B+ ewes, 5/6 ewes did not have any follicles that expressedCARTPT, and no CART peptide was detected in any follicle examined. Expression pattern ofCYP19A1differed between I+B+ and ++ ewes with an increased percentage of small and medium follicles (3 to <4.5 mm diameter) but decreased percentage of large follicles (≥4.5 mm diameter) expressingCYP19A1in the I+B+ ewes. Many of the large follicles from the I+B+ ewes appeared non-functional and expression ofLHCGR,FSHR,CYP17A1andCYP19A1was less than that observed in ++ ewes. Expression ofFSHRandCYP17A1was not different between groups in small and medium follicles, butLHCGRexpression was approximately double in I+B+ ewes compared to that in ++ ewes. Thus, ewes with high ovulation rates had a distinct pattern of expression ofCARTPTmRNA and protein compared to ewes with normal ovulation rates, providing evidence for CART being important in the regulation of ovulation rate.

scholarly journals Tiling Array Analysis of UV Treated Escherichia coli Predicts Novel Differentially Expressed Small Peptides

PLoS ONE ◽  
2010 ◽  
Vol 5 (12) ◽  
pp. e15356 ◽  
Author(s):  
Gard O. S. Thomassen ◽  
Ragnhild Weel-Sneve ◽  
Alexander D. Rowe ◽  
James A. Booth ◽  
Jessica M. Lindvall ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Tiling Array ◽  
Differentially Expressed ◽  
Small Peptides ◽  
Array Analysis

115 Exosome-mediated microRNA expression profile in follicular fluid of metabolically divergent postpartum cows

2019 ◽  
Vol 31 (1) ◽  
pp. 183
Author(s):  
T. Hailay ◽  
M. Hoelker ◽  
S. Gebremedhn ◽  
F. Rings ◽  
M. M. Saeed-Zidane ◽  
...  
Keyword(s):  
Energy Balance ◽  
Follicular Fluid ◽  
Mirna Expression ◽  
Cell Communication ◽  
Negative Energy ◽  
Negative Energy Balance ◽  
Differentially Expressed ◽  
Metabolic Status ◽  
Lactation Period ◽  
Down Regulation

Most high-milking cows enter a state of negative energy balance during the early lactation period. This phenomenon disturbs the metabolic status of the follicular fluid microenvironment, resulting in delayed ovulation. Cell-to-cell communication between the oocyte and the surrounding cells is crucial during folliculogenesis. Exosomes, evolutionarily conserved cargo molecules (30-150nm in diameter) carrying RNA and proteins, are known to be involved in cell-to-cell communication. Here, we aimed to investigate the association between postpartum metabolic status and the expression of exosomal microRNA (miRNA) in follicular fluid of Holstein-Friesian cows. For this, follicular fluid was collected from antral follicles (&gt;8mm in diameter) using ovum pickup procedure from cows (n=30) on a weekly basis between weeks 5 and 10 postpartum. Follicular fluid collected from heifers (n=8) was used as a control. The energy status of each cow was assessed based on the blood metabolite (nonesterified fatty acids and β-hydroxybutyrate) concentration, body weight curve, and overall energy balance determined by dry matter intake. Afterwards, cows were categorized as early negative and late positive (cows show negative energy balance at early weeks and recovered at late weeks postpartum), always negative (cows did not recover until 15 weeks postpartum), and always positive (cows did not enter in to a state of negative energy balance). Following this, exosomes were isolated from pooled samples from each animal category using ultracentrifugation, and their morphology and size was characterised using electron microscopy and nanosight, respectively. Exosomal total RNA enriched with miRNA was isolated using an exosomal RNA isolation kit. Next-generation sequencing of miRNA was performed using Illumina NextSEqn 500 (Illumina Inc., San Diego, CA, USA). MicroRNAs with a fold change ≥2, P-value &lt;0.05, and a false discovery rate of &lt;0.1 were considered differentially expressed. The results showed that a total of 356 known and 156 novel miRNA were identified across samples. Differential expression analysis of miRNA between always-negative cows versus always-positive cows revealed down-regulation of all 6 differentially expressed miRNA, including bta-miR-451, bta-miR-132, and bta-miR-2285. Similarly, down-regulation of 14 miRNA, including bta-miR-20b, bta-miR-363, bta-miR-132, and bta-miR-451, and up-regulation of 3 miRNA was observed in always-negative cows compared to heifers. Furthermore, the target prediction analysis of the down-regulated miRNA have been shown to be involved in regulating different pathways including transforming growth factor-β signalling, cell cycle, hippo signalling, forkhead box O signalling, and endometrial cancer, among others. In conclusion, the results revealed that although negative energy balance in postpartum dairy cows suppressed exosomal miRNA expression in follicular fluid, the opposite was observed in metabolically unstressed cows. This divergence of exosome-mediated miRNA expression in the follicular fluid of metabolically stressed cows could be associated with the reduced fertility of those cows.

scholarly journals Identification of Candidate Genes and Pathways in Dexmedetomidine-Induced Cardioprotection in the Rat Heart by Bioinformatics Analysis

2019 ◽  
Vol 20 (7) ◽  
pp. 1614 ◽  
Author(s):  
Yusuke Yoshikawa ◽  
Naoyuki Hirata ◽  
Hirofumi Terada ◽  
Yasuaki Sawashita ◽  
Michiaki Yamakage
Keyword(s):  
Map Kinase ◽  
Candidate Genes ◽  
Phosphatase Activity ◽  
Bioinformatics Analysis ◽  
Ischemia Reperfusion ◽  
Differentially Expressed ◽  
P53 Pathway ◽  
Rat Hearts ◽  
Significant Pathway ◽  
Adrenergic Receptor Agonist

Dexmedetomidine (DEX), a highly selective alpha2 adrenergic receptor agonist, directly protects hearts against ischemia/reperfusion (I/R) injury. However, the detailed mechanism has not been fully elucidated. We studied differentially expressed mRNAs and miRNAs after DEX administration in rat hearts by comprehensive analysis. Additionally, bioinformatics analysis was applied to explore candidate genes and pathways that might play important roles in DEX-induced cardioprotection. The results of microarray analysis showed that 165 mRNAs and 6 miRNAs were differentially expressed after DEX administration. Through bioinformatics analysis using differentially expressed mRNAs, gene ontology (GO) terms including MAP kinase tyrosine/serine/threonine phosphatase activity and pathways including the p53 pathway were significantly enriched in the down-regulated mRNAs. Dusp1 and Atm were associated with the GO term of MAP kinase tyrosine/serine/threonine phosphatase activity and the p53 pathway, respectively. On the other hand, no significant pathway was found in the target mRNAs of deregulated miRNAs. The results indicated some possible key genes and pathways that seem to be of significance in DEX-induced cardioprotection, although miRNAs seem to be unlikely to contribute to cardioprotection induced by DEX.

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