Phosphorylation culture in vitro oocyte maturation techniques. Medical search. Frequent questions

MicroRNAs (miRNAs) are small non-encoding RNAs that actively regulate biological and physiological processes, and play an important role in regulating gene expression in all cells, especially in most animal cells, including oocytes and embryos. (bvsalud.org)
Although assisted reproductive techniques (ART) have helped to solve many infertility problems, they cause changes in the expression of miRNA and genes in oocytes and preimplantation embryos, and the effect of these changes on the future of offspring is unknown, and has caused concerns. (bvsalud.org)
To achieve this, cells of in vitro produced bovine embryos were traced from the 2-cell stage to the blastocyst stage. (bioone.org)
As a result of a lower number of oocytes retrieved, there are fewer embryos to select and transfer, and subsequently these patients have lower pregnancy rates per transfer and lower cumulative pregnancy rates per started cycle compared with normal responders. (biomedcentral.com)
After that, we examined the result of cultured conditioned mass media (CM) of IVF embryos on splenic immune system cells and Madin-Darby bovine kidney (MDBK) cells with stably presented ISG15 promoter-reporter constructs. (bioskinrevive.com)
For in vivo test: In vitro lifestyle and embryo freezing After IVF, oocytes encircled with cumulus cells had been placed in fresh new TCM-199 moderate, as well as the embryos had been co-cultured with cumulus cells, as previously defined [21]. (bioskinrevive.com)
The two enrichment strategies allow probing of methyl-CpG protein interactions in early vertebrate oocytes and embryos. (biomedcentral.com)
In contrast to the semi-dry standard culture method, this setting corresponded more to the in vivo conditions of gastrulating/neurulating rabbit embryos in the uterus. (studylibde.com)
Conversely, in standard-cultured embryos showing altered LR marker gene expression, maturation of the PNC was impaired leading to a disturbance of leftward flow. (studylibde.com)
When the flow was blocked during culture by methylcellulose-containing medium embryos displayed altered LR marker gene expression in a very high proportion. (studylibde.com)

Xenopus oocytes are encompassed by a layer of follicular cells that contribute to oocyte growth and meiosis in relation to oocyte maturation. (bvsalud.org)
Here, we investigated Xenopus follicular cell function using oocyte signaling and heterologous-expressing capabilities. (bvsalud.org)
Endogenous MeCP2 and MBD3 were precipitated from Xenopus oocyte extracts and conditions for methylation-specific binding were optimized. (biomedcentral.com)
an investigation of their specific localization in human brain and their influence on CaV2.1 voltage-dependent calcium channels expressed in Xenopus oocytes. (biomedcentral.com)
In addition, we investigated whether human γ 2 or γ 4 could modulate the electrophysiological properties of a neuronal VDCC complex transiently expressed in Xenopus oocytes. (biomedcentral.com)
When co-expressed in Xenopus oocytes with a Ca V 2.1/β 4 VDCC complex, either in the absence or presence of an α2δ 2 subunit, neither γ 2 nor γ 4 significantly modulated the VDCC peak current amplitude, voltage-dependence of activation or voltage-dependence of steady-state inactivation. (biomedcentral.com)
Here we used two-electrode voltage clamp electrophysiology to compare the actions of CBD with those of the major central endocannabinoid, 2-arachidonoyl glycerol (2-AG) on human recombinant GABAA receptors (synaptic α1-6βg2 and extrasynaptic α4β2δ) expressed on Xenopus oocytes. (researchgate.net)

After either in vivo or in vitro capacitation the sperm must undergo the final maturation step, activation, involving the acrosome reaction. (wikipedia.org)
Components and OPTIONS FOR in vivo test: In vitro maturation and fertilization maturation and fertilization had been carried out as described [21] previously. (bioskinrevive.com)
This review focuses on the differential responses of cell types to ATZ toxicity, testicular effects of ATZ in both in vitro and in vivo models and chemopreventive strategies, so as to highlight the current state of the art on the toxicological outcomes of ATZ exposure in several experimental model systems. (frontiersin.org)
However, DFMO had been mainly without effect of these inside vitro or even in vivo assays. (hsp90-inhibitors.com)

Maturation of the mammalian oocyte is a complex process involving internal checkpoints and bidirectional communication with the surrounding cumulus cells. (biomedcentral.com)
The mammalian oocyte extracellular matrix known as the zona pellucida (ZP) acts as a barrier to accomplish sperm fusion with the female gamete. (biomedcentral.com)

In addition, we determined that oocytes surrounded by their follicular cells expressing EGFR-Grb7 exhibit normal meiotic resumption. (bvsalud.org)
The oocyte must maintain arrest at the diplotene stage until meiotic resumption occurs. (biomedcentral.com)

For purposes of in vitro fertilization, capacitation occurs by incubating spermatozoa that have either undergone ejaculation or have been extracted from the epididymis and incubated in a defined medium for several hours. (wikipedia.org)
citation needed] Because assisted reproductive technologies, or ARTs, such as in vitro fertilization (IVF) or intrauterine insemination (IUI) require the induction of sperm cell capacitation outside of normal biological parameters, numerous methods have been developed to induce this process in mammalian sperm cells. (wikipedia.org)
For animal in vitro fertilization, caffeine at 5 mM concentration is a strong inducer of sperm capacitation in vitro. (wikipedia.org)
Poor ovarian responders (POR) are women undergoing in-vitro fertilization who respond poorly to ovarian stimulation, resulting in the retrieval of lower number of oocytes, and subsequently lower pregnancy rates. (biomedcentral.com)
Following germinal vesicle break down (GVBD) the oocyte is transcriptionally quiescent until fertilization and activation of the embryonic genome, occurring at the four-cell stage of development in the pig [ 9 ]. (biomedcentral.com)
During mammalian fertilization, sperm should be able to reach the cumulus-oocyte complex, get through the hyaluronic matrix of the cumulus and penetrate the zona pellucida (ZP) in order to fuse with the egg. (biomedcentral.com)

Lyons VG, Payne NL, McCoy CE (2018) Optimization techniques for miRNA expression in low frequency immune cell populations. (hudson.org.au)

However, the effects of the interaction between follicular cells and the oocyte surface on meiotic processes are unclear. (bvsalud.org)
We found that oocytes deprotected from their surrounding layer of follicular cells and expressing the epidermal growth factor (EGF) receptor (EGFR) and the Grb7 adaptor undergo accelerated prophase I to metaphase II meiosis progression upon stimulation by EGF. (bvsalud.org)
Furthermore, an O-GlcNAcylation increase (by inhibition of O-GlcNAcase), the glycosidase that removes O-GlcNAc moieties, or decrease (by inhibition of OGT) amplifies oocyte spindle defects when follicular cells are absent highlighting a control of the meiotic spindle by the OGT-O-GlcNAcase duo. (bvsalud.org)
Growth of the dominant follicle encompasses enlargement of the oocyte, replication of follicular cells, and formation and expansion of a fluid-filled follicular antrum or cavity, providing a specialized microenvironment for the development of oocytes. (biomedcentral.com)

The follicular fluid (FF) provides a crucial microenvironment for the proper development of follicles and oocytes through tightly controlled metabolism and cell signaling. (biomedcentral.com)
Oocytes were collected from aspirated porcine tertiary follicles. (biomedcentral.com)

At least STAT5 requires glycosylation at threonine 92 for strong STAT5 tyrosine phosphorylation. (statsignaling.com)

Next, to find out regardless of whether pre or post synaptic professional tein synthesis is necessary for NT 3 mediated long term synaptic modulation, we expressed GyrB PKR in either spinal neurons or myocytes applying precisely the same embryo injection techniques described over. (narturalproduct.com)
Establishment of a new in vitro Culture System and functional Analysis of Sonic Hedgehog and FGF8 in the Determination of Laterality in the Rabbit Embryo Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften (Dr. rer. (studylibde.com)
In this new method, the so-called ring culture, a medium-filled plastic ring was placed upon the extraembryonic tissue of the explanted embryo. (studylibde.com)

Cultures have been incubated with NT 3 for two days with or without coumermycin as indicated, At naive synapses, coumermycin treatment method didn't impact basal synaptic transmission nor prevent the long term poten tiating result of NT 3, Expres sion of GyrB PKR in either presynaptic spinal neurons or postsynaptic muscle cells devoid of coumermycin treat ment didn't alter the long lasting impact of NT three. (narturalproduct.com)
Among those, several candidate genes have been shown to control the early patterning and/or the late synaptic maturation of specific neuronal subpopulations controlling the balance between excitation and inhibition in the developing cortex and cerebellum. (neurotransmitter.net)

Excessive accumulation of adipose tissue in cultured fish is an outstanding problem in aquaculture. (biomedcentral.com)
We have previously shown that fish primary preadipocytes differentiate into mature adipocytes in vitro and that these cells represent a very helpful model system to study adipose tissue development in fish [ 5 , 6 ]. (biomedcentral.com)
FLIM-FRET analysis of protein-protein interactions showed that PLIN5 S155 phosphorylation regulates PLIN5 interaction with adipose triglyceride lipase at the lipid droplet, but not with α-β hydrolase domain-containing 5. (uci.edu)

Inhibition of nuclear meiotic maturation via IBMX significantly increased MIR21 and decreased its target, PDCD4. (biomedcentral.com)

This unusual maturation unravels atypical spindle formation but is rescued by inhibiting integrin ß1 or Grb7 binding to the EGFR. (bvsalud.org)
These oocytes are protected from abnormal meiotic spindle formation through the recruitment of O-GlcNAcylated Grb7, and OGT (O-GlcNAc transferase), the enzyme responsible for O-GlcNAcylation processes, in the integrin ß1-EGFR complex. (bvsalud.org)
Works in various organisms have revealed that the kinase is involved in centrosome separation, duplication and maturation as well as in bipolar spindle assembly and stability. (lookformedical.com)

In summary, our study provides further insight into the role of the follicular cell layer in oocyte meiosis progression. (bvsalud.org)
In affected ovaries, oxygenation of follicular fluid is low, ultrastructural disturbances especially of mitochondria, occur in granulosa cells and oocytes, and considerable disturbances of meiosis occur. (biomedcentral.com)

Human oocyte maturation arrest represents one of the severe conditions for female patients with primary infertility. (bvsalud.org)
In this study, by whole-exome sequencing (WES), we identified homozygous and compound heterozygous MAD2L1BP variants in three families with female patients diagnosed with primary infertility owing to oocyte metaphase I (MI) arrest. (bvsalud.org)
Together, our studies identified and characterized novel biallelic variants in MAD2L1BP responsible for human oocyte maturation arrest at MI, and thus prompted new therapeutic avenues for curing female primary infertility. (bvsalud.org)
Oocyte arrest at the diplotene stage is maintained in part through the activity of phosphodiesterase enzymes that catalyze the hydrolysis of cyclic adenosine monophosphate (cAMP) into AMP [ 1 , 2 ]. (biomedcentral.com)

A group of enzymes that catalyzes the phosphorylation of serine or threonine residues in proteins, with ATP or other nucleotides as phosphate donors. (lookformedical.com)

We sought to identify PKA phosphorylation sites in PLIN5 and assess their functional relevance in cultured cells and the livers of mice. (uci.edu)

Ectopic kinase overexpression in any culture cell line leads to polyploidy and centrosome amplification. (lookformedical.com)
Perilipin 5 (PLIN5) is a lipid-droplet-associated protein that coordinates intracellular lipolysis in highly oxidative tissues and is thought to regulate lipid metabolism in response to phosphorylation by protein kinase A (PKA). (uci.edu)

Bovine serum albumin is typically used for in vitro animal studies, and human serum albumin (HSA) is used in human sperm capacitation induction. (wikipedia.org)
Matured oocytes had been inseminated with frozen-thawed semen from order MK-0822 a Japanese Dark bull (altered to 2 107 cells/ml) for 5 h at 38.5C within a humidified atmosphere with 5% CO2 in the surroundings, in 1 ml of BO solution containing 10 mg/ml bovine serum albumin and 10 g/ml heparin. (bioskinrevive.com)

They include metabolic products highly associated with ovarian function - amino acids for regulating pH and osmolarity, lipids such fatty acids and cholesterols for oocyte maturation, and glucocorticoids for ovarian steroidogenesis. (biomedcentral.com)

Adding heparin to capacitation inducing medium mimics the secretion of heparin-like gycosaminoglycans (GAGs) near the oocyte and initiates the acrosome reaction. (wikipedia.org)

The expression of miRNAs at the right time and place is crucial for the oocyte's maturation and the embryo's subsequent development. (bvsalud.org)
Expression of phosphorylation-defective PLIN5 S155A in Plin5 null cells resulted in decreased rates of lipolysis and triglyceride-derived fatty acid oxidation. (uci.edu)

Proliferation cluster comprised 1028 genes up-regulated from days 3 to 8 of culture meanwhile the differentiation cluster was characterized by 2140 induced genes from days 15 to 21. (biomedcentral.com)

Germinal vesicle breakdown (GVBD) occurs during oocyte meiotic maturation, a period when transcriptional processes are virtually inactive. (biomedcentral.com)

The mammalian genome comprises nuclear DNA (nDNA) derived from both parents and mitochondrial DNA (mtDNA) that is maternally inherited and encodes essential proteins required for oxidative phosphorylation. (regenerativemedicine.net)

Bicarbonate is a vital component of capacitation-inducing media, as it is co-transported into the cytosol where it activates soluble adenylyl cyclase (sAC) as well as acts as a pH buffer necessary to prevent decreasing the pH in the culture, a necessary addition when incubating cells at 5% CO2 as is generally used although not required. (wikipedia.org)
72 hours) to C8- ceramide (5 μg/ml culture medium), a cell-permeable analog of ceramide, enhanced MDR1 mRNA levels by 3- and 5-fold in T47D and inMDA-MB-435 cells, respectively, but did not affect MCF-7 cells.D-erythro-sphingosine exposure (5 μg/ml, 72 hr) increased MDR1 mRNA levels in MDA-MB-435 cells by 3.5-fold. (sphingolipidclub.com)
Acute exposure ofMCF-7 and MDA-MB-231 cells to C8-glucosylceramide (10 μg/ml culture medium), a cell-permeable analog of glucosylceramide, increased MDR1 mRNA levels by 2- and 4-fold, respectively. (sphingolipidclub.com)
In cultured primary fibroblasts and cancer cells, the chemotherapeutic drug doxorubicin causes mtDNA damage and release, which leads to cGAS STING dependent ISG activation. (regenerativemedicine.net)

The traditional methods to perform in vitro capacitation are: Simple wash: this method only eliminates seminal plasma, it does not select the best spermatozoa. (wikipedia.org)

Injection of a MIR21 mimic increased oocyte maturation rate. (biomedcentral.com)

When in vitro experimental models are exposed to ATZ, they are faced with increased levels of reactive oxygen species (ROS), cytotoxicity and decreased growth rate at dosages that may vary with cell types. (frontiersin.org)

Today, as research techniques become more refined and the evidence generated more complex, silos of knowledge are generated and experts find it difficult to cross between sub-disciplines. (biomedcentral.com)

Our results indicate MIR21 is active and important during meiotic maturation of the oocyte. (biomedcentral.com)

Understanding the biological functions of miRNAs in frozen maturated oocytes may provide a better understanding of embryonic development and a comparison of fertility conservation in female mammals. (bvsalud.org)
The inability to transcribe mRNA during this stage of development and the probable necessity for post-transcriptional gene regulation (PTGR) suggests an important role for non-coding RNA in the maturing oocyte. (biomedcentral.com)
Therefore, the transcriptomic profile at different time points (days 3, 8, 15 and 21) along primary culture development of rainbow trout preadipocytes has been investigated using an Agilent trout oligo microarray. (biomedcentral.com)
It therefore facilitated stable development of laterality in most cases also when presomite stages were taken into culture. (studylibde.com)
It is well accepted that adverse life events occurring early in development may alter the correct program of brain maturation leading to enhanced vulnerability to neuropsychiatric disorders. (researchgate.net)

Of particular interest is understanding the role of non-coding RNA in regulating meiotic checkpoints in the oocyte, particularly that of microRNA-21 (MIR21). (biomedcentral.com)

Using in vitro and cellular techniques, we present a mVP40 assembly model highlighting two distinct oligomerization interfaces: the (N-terminal domain (NTD) and C-terminal domain (CTD)) in mVP40. (uci.edu)

In patients designated as "poor responders," so-called due to poor response to ovarian stimulation given during IVF workup, the limited number of obtained oocytes remains the major problem in optimizing the live birth rates [ 3 ]. (biomedcentral.com)
Since recent clinical studies have indicated that dehydroepiandrosterone (DHEA) supplementation may reverse the problem of oocyte quality, this review of the literature was undertaken in an attempt to find an explanation of why this is effective? (biomedcentral.com)

To characterize the effect of artificially increasing MIR21 on oocyte competence without inhibiting GVBD, a MIR21 mimic, scrambled microRNA negative control, or nuclease free water was micro-injected into denuded oocytes at 21 h of IVM. (biomedcentral.com)
Perilipin 5 S155 phosphorylation by PKA is required for the control of hepatic lipid metabolism and glycemic control. (uci.edu)

Folliculated oocytes can be forced to adopt an abnormal phenotype and exclusive Grb7 Y338 and Y188 phosphorylation instead of O-GlcNAcylation under integrin activation. (bvsalud.org)

A recent meta-analysis study found removal of oocytes from the disrupted endogenous steroidal environment of PCOS women and maturing them in vitro helps achieve better conception rates [ 12 ]. (biomedcentral.com)

Non-mammalian spermatozoa do not require this capacitation step and are ready to fertilize an oocyte immediately after release from the male. (wikipedia.org)
In the farming setting, temperature outcomes in which probably interrupt food accessibility might be more essential as compared to outcomes IKK-16 in vitro immediately affecting your success of Whitened Stork children. (hsp90-inhibitors.com)

These specific advantages of the rabbit were employed in this study to further examine the role of two central determinants of laterality, namely Sonic hedgehog (Shh) and FGF8, and also extended by the design of a new in vitro culture technique. (studylibde.com)

3) that were divided into two major clusters corresponding to the main phases observed during the preadipocyte culture: proliferation and differentiation. (biomedcentral.com)

Euglena gracilis can grow in the mixed culture containing Cladosporium westerdijkiae, Lysinibacillus boronitolerans and Pseudobacillus badius without the addition of vitamins B1 and B12. (cuni.cz)

Relative abundance of mature MIR21 was quantified at 0, 8, 16, 24, 32, and 42 h of in vitro (IVM) with or without treatment with 3-isobutyl-1-methylxanthine (IBMX). (biomedcentral.com)

We establish a pH dependant oligomerisation pathway forming tetrameric DEC-205 using solution-based techniques and ultimately solved the 4.9 Å cryo-EM structure of the DEC-205 tetramer to identify the unfurling of the second lectin ring which enables tetramer formation. (uci.edu)

Anatomy2

Organisms1

Analytical, Diagnostic and Therapeutic Techniques and Equipment2

Phenomena and Processes3

Embryos10

Xenopus7

Vivo4

Mammalian oocyte2

Meiotic resumption2

Fertilization6

MiRNA1

Follicular cells4

Follicles2

STAT51

Embryo3

Synaptic2

Adipose3

Inhibition1

Spindle3

Meiosis2

Arrest4

Threonine1

Functional1

Kinase2

Bovine2

Lipids1

Heparin1

Expression2

Genes1

Occurs1

Proteins1

Cells4

Methods1

Injection1

Species1

Complex1

Results1

Development5

Understanding1

Present1

Problem2

Control2

Activation1

Found1

Immediately2

Specific1

Main1

Addition1

Treatment1

Formation1