ABSTRACT
Chromosome testing strategies, such as preimplantation genetic testing for aneuploidy (PGT-A), improve initial IVF outcomes by avoiding unwitting transfer of aneuploid embryos in morphology-based selection practices. Newer technologies have revealed that some embryos may appear to have intermediate whole chromosome (or parts of a chromosome termed segmental) copy number results suggesting trophectoderm mosaicism. An embryo with a trophectoderm mosaic-range result may be the only option for transfer for some patients. Recent data suggest that such mosaic embryos can be transferred without added risk of abnormal birth outcomes but may be associated with increased implantation failure and miscarriage rates, with higher values of mosaicism appearing to be less favourable for producing good outcomes. In this Position Statement, we provide guidance to laboratories, clinics, clinicians and counsellors to assist in discussions on the utility and transfer of mosaic embryos.
Subject(s)
Preimplantation Diagnosis , Aneuploidy , Blastocyst , Embryo Transfer , Female , Genetic Testing/methods , High-Throughput Nucleotide Sequencing/methods , Humans , Mosaicism , Pregnancy , Preimplantation Diagnosis/methodsABSTRACT
PURPOSE: In men, obesity may lead to poor semen parameters and reduced fertility. However, the causative links between obesity and male infertility are not totally clear, particularly on a molecular level. As such, we investigated how obesity modifies the human sperm proteome, to elucidate any important implications for fertility. METHODS: Sperm protein lysates from 5 men per treatment, classified as a healthy weight (body mass index (BMI) ≤ 25 kg/m2) or obese (BMI ≥ 30 kg/m2), were FASP digested, submitted to liquid chromatography tandem mass spectrometry, and compared by label-free quantification. Findings were confirmed for several proteins by qualitative immunofluorescence and a quantitative protein immunoassay. RESULTS: A total of 2034 proteins were confidently identified, with 24 proteins being significantly (p < 0.05) less abundant (fold change < 0.05) in the spermatozoa of obese men and 3 being more abundant (fold change > 1.5) compared with healthy weight controls. Proteins with altered abundance were involved in a variety of biological processes, including oxidative stress (GSS, NDUFS2, JAGN1, USP14, ADH5), inflammation (SUGT1, LTA4H), translation (EIF3F, EIF4A2, CSNK1G1), DNA damage repair (UBEA4), and sperm function (NAPA, RNPEP, BANF2). CONCLUSION: These results suggest that oxidative stress and inflammation are closely tied to reproductive dysfunction in obese men. These processes likely impact protein translation and folding during spermatogenesis, leading to poor sperm function and subfertility. The observation of these changes in obese men with no overt andrological diagnosis further suggests that traditional clinical semen assessments fail to detect important biochemical changes in spermatozoa which may compromise fertility.
Subject(s)
Fertility/genetics , Obesity/genetics , Proteome/genetics , Spermatogenesis/genetics , Adult , Body Mass Index , Female , Humans , Infertility, Male/complications , Infertility, Male/genetics , Infertility, Male/metabolism , Infertility, Male/pathology , Male , Obesity/complications , Obesity/pathology , Oxidative Stress/genetics , Proteome/metabolism , Semen Analysis , Sperm Count , Sperm Motility/genetics , Spermatozoa/metabolism , Spermatozoa/pathologyABSTRACT
This paper reviews 28 papers or commentaries published in Journal of Clinical Monitoring and Computing in 2018 and 2019, within the field of respiration. Papers were published covering endotracheal tube cuff pressure monitoring, ventilation and respiratory rate monitoring, lung mechanics monitoring, gas exchange monitoring, CO2 monitoring, lung imaging, and technologies and strategies for ventilation management.
Subject(s)
Respiration , Biomechanical Phenomena , Carbon Dioxide/metabolism , Electric Impedance , Humans , Intubation, Intratracheal , Lung/diagnostic imaging , Lung/physiology , Monitoring, Physiologic , Pressure , Pulmonary Gas Exchange , Pulmonary Ventilation , Respiration, Artificial , Respiratory Rate , TomographySubject(s)
Chimera/embryology , Embryo Transfer/standards , Mosaicism , Preimplantation Diagnosis/standards , Embryo Transfer/methods , Female , Humans , Internationality , Pregnancy , Pregnancy Outcome/genetics , Preimplantation Diagnosis/methods , Reproductive Techniques, Assisted/standards , Societies, Medical/standardsABSTRACT
PURPOSE: The aim of this study was to evaluate the incidence of an inter-chromosomal effect (ICE) in blastocyst-stage embryos from carriers of balanced chromosome inversions. METHODS: Infertility patients (n = 52) with balanced inversions (n = 66 cycles), and maternal age-matched controls that concurrently cycled (n = 66), consented to an IVF cycle with preimplantation genetic testing for aneuploidy (PGT-A). Blastocyst-stage embryos underwent trophectoderm biopsy for PGT-A with only euploid blastocysts transferred in a subsequent frozen embryo transfer. Subtypes of inversions were included in aggregate: paracentric/pericentric, polymorphic/non-polymorphic, male/female carriers, and varying inversion sizes. RESULTS: The incidence of aneuploidy was not significantly higher for the inversion patients compared to the controls (inversion = 48.8% vs. control = 47.2% ns). Following euploid blastocyst transfer, there were excellent live birth outcomes. CONCLUSIONS: Carriers of balanced chromosome inversions did not exhibit higher aneuploidy rates for chromosomes that were not involved in the inversion compared to maternal age-matched controls, signifying the absence of an inter-chromosomal effect for this data set. These results provide the largest investigation of blastocyst embryos regarding the debated existence of an ICE resulting from the presence of an inversion during meiosis. However, further studies are warranted to investigate an ICE among inversions subtypes that were outside the scope of this study.
Subject(s)
Chromosome Inversion/genetics , Embryonic Development/genetics , Fertilization in Vitro , Infertility/genetics , Adult , Aneuploidy , Blastocyst/pathology , Comparative Genomic Hybridization , Embryo Transfer , Female , Genetic Testing , Humans , Infertility/physiopathology , Maternal Age , Pregnancy , Pregnancy Rate , Preimplantation DiagnosisABSTRACT
AIMS: To evaluate the effectiveness of automated symptom and side effect monitoring on quality of life among individuals with symptomatic diabetic peripheral neuropathy. METHODS: We conducted a pragmatic, cluster randomized controlled trial (July 2014 to July 2016) within a large healthcare system. We randomized 1834 primary care physicians and prospectively recruited from their lists 1270 individuals with neuropathy who were newly prescribed medications for their symptoms. Intervention participants received automated telephone-based symptom and side effect monitoring with physician feedback over 6 months. The control group received usual care plus three non-interactive diabetes educational calls. Our primary outcomes were quality of life (EQ-5D) and select symptoms (e.g. pain) measured 4-8 weeks after starting medication and again 8 months after baseline. Process outcomes included receiving a clinically effective dose and communication between individuals with neuropathy and their primary care provider over 12 months. Interviewers collecting outcome data were blinded to intervention assignment. RESULTS: Some 1252 participants completed the baseline measures [mean age (sd): 67 (11.7), 53% female, 57% white, 8% Asian, 13% black, 20% Hispanic]. In total, 1179 participants (93%) completed follow-up (619 control, 560 intervention). Quality of life scores (intervention: 0.658 ± 0.094; control: 0.653 ± 0.092) and symptom severity were similar at baseline. The intervention had no effect on primary [EQ-5D: -0.002 (95% CI -0.01, 0.01), P = 0.623; pain: 0.295 (-0.75, 1.34), P = 0.579; sleep disruption: 0.342 (-0.18, 0.86), P = 0.196; lower extremity functioning: -0.079 (-1.27, 1.11), P = 0.896; depression: -0.462 (-1.24, 0.32); P = 0.247] or process outcomes. CONCLUSIONS: Automated telephone monitoring and feedback alone were not effective at improving quality of life or symptoms for people with symptomatic diabetic peripheral neuropathy. TRIAL REGISTRATION: ClinicalTrials.gov (NCT02056431).
Subject(s)
Diabetic Neuropathies/therapy , Monitoring, Physiologic/methods , Primary Health Care , Quality of Life , Aged , Cluster Analysis , Diabetic Neuropathies/physiopathology , Diabetic Neuropathies/psychology , Female , Humans , Male , Middle Aged , Outcome Assessment, Health Care , Practice Patterns, Physicians'ABSTRACT
This paper reviews 32 papers or commentaries published in Journal of Clinical Monitoring and Computing in 2016, within the field of respiration. Papers were published covering airway management, ventilation and respiratory rate monitoring, lung mechanics and gas exchange monitoring, in vitro monitoring of lung mechanics, CO2 monitoring, and respiratory and metabolic monitoring techniques.
Subject(s)
Monitoring, Physiologic/methods , Periodicals as Topic , Respiration , Respiratory Rate , Animals , Capnography , Carbon Dioxide/chemistry , Clinical Trials as Topic , Electric Impedance , Humans , Lung/physiology , Monitoring, Physiologic/instrumentation , Oximetry , Pulmonary Gas Exchange , Respiration, Artificial , Signal Processing, Computer-AssistedABSTRACT
This paper reviews 16 papers or commentaries published in Journal of Clinical Monitoring and Computing in 2016, within the field of respiration. Papers were published covering peri- and post-operative monitoring of respiratory rate, perioperative monitoring of CO2, modeling of oxygen gas exchange, and techniques for respiratory monitoring.
Subject(s)
Monitoring, Physiologic/instrumentation , Monitoring, Physiologic/methods , Oximetry/methods , Respiration , Respiratory Rate , Algorithms , Animals , Capnography , Carbon Dioxide , Electric Impedance , Electrocardiography , Humans , Postoperative Period , Pulmonary Gas Exchange , Respiration, ArtificialABSTRACT
This paper reviews 17 papers or commentaries published in Journal of Clinical Monitoring and Computing in 2015, within the field of respiration. Papers were published covering monitoring and training of breathing, monitoring of gas exchange, hypoxemia and acid-base, and CO2 monitoring.
Subject(s)
Lung Diseases/diagnosis , Periodicals as Topic/trends , Polysomnography/trends , Respiration Disorders/diagnosis , Respiratory Function Tests/trends , Humans , Lung Diseases/prevention & control , Polysomnography/instrumentation , Polysomnography/methods , Respiration Disorders/prevention & control , Respiratory Function Tests/instrumentation , Respiratory Function Tests/methodsABSTRACT
If dark energy, which drives the accelerated expansion of the universe, consists of a light scalar field, it might be detectable as a "fifth force" between normal-matter objects, in potential conflict with precision tests of gravity. Chameleon fields and other theories with screening mechanisms, however, can evade these tests by suppressing the forces in regions of high density, such as the laboratory. Using a cesium matter-wave interferometer near a spherical mass in an ultrahigh-vacuum chamber, we reduced the screening mechanism by probing the field with individual atoms rather than with bulk matter. We thereby constrained a wide class of dark energy theories, including a range of chameleon and other theories that reproduce the observed cosmic acceleration.
ABSTRACT
Ammonium is generated in culture media by the spontaneous deamination of amino acids at 37â°C and through the metabolism of amino acids by human embryos. The appearance of ammonium is a time-dependent phenomenon and can compromise embryo physiology, development and viability. In this study, the effects of a gradient of ammonium on the development, metabolism and transcriptome of human and mouse embryos were investigated. Pronucleate oocytes were cultured in the presence of an ammonium gradient that mimicked the spontaneous deamination of Eagle's amino acids together with 1 mM glutamine. All embryos were cultured in sequential media G1/G2 at 5% O2, 6% CO2 and 89% N2. Human embryo metabolism was assessed through a non-invasive fluorometric analysis of pyruvate consumption. Transcriptome analysis was performed on the resultant blastocysts from both species using a microarray technology. Embryo development prior to compaction was negatively affected by the presence of low levels of ammonium in both species. Human embryo metabolism was significantly inhibited after just 24 and 48 h of culture. Transcriptome analysis of blastocysts from both species revealed significantly altered gene expression profiles, both decreased and increased. Functional annotation of the altered genes revealed the following over represented biological processes: metabolism, cell growth and/or maintenance, transcription, cell communication, transport, development and transcription regulation. These data emphasize the enhanced sensitivity of the cleavage-stage embryo to its environment and highlight the requirement to renew culture media at frequent intervals in order to alleviate the in vitro induced effects of ammonium build-up in the environment surrounding the embryo.
Subject(s)
Ammonium Compounds/adverse effects , Embryo Culture Techniques , Embryo, Mammalian/drug effects , Embryonic Development/drug effects , Gene Expression Regulation, Developmental/drug effects , Animals , Embryo, Mammalian/metabolism , Female , Humans , Metabolism/drug effects , Mice , PregnancyABSTRACT
STUDY QUESTION: When a chromosome aneuploidy is detected in the first polar body and a reciprocal loss or gain of the same chromosome is detected in the second polar body, is the resulting embryo usually aneuploid for that chromosome? SUMMARY ANSWER: When reciprocal aneuploidy occurs in polar bodies, the resulting embryo is usually normal for that chromosome, indicating that premature separation of sister chromatids (PSSC)-not non-disjunction-likely occurred in meiosis I. WHAT IS KNOWN ALREADY: Single-nucleotide polymorphism-based microarray analysis can be used to accurately determine the chromosomal status of polar bodies and embryos. Sometimes, the only abnormality found is a reciprocal gain or loss of one or two chromosomes in the two polar bodies. Prediction of the status of the resulting embryo in these cases is problematic. STUDY DESIGN, SIZE, DURATION: Blinded microarray analysis of previously diagnosed aneuploid embryos that had reciprocal polar body aneuploidy. MATERIALS, SETTING, METHODS: IVF cycles were performed between 2008 and 2011 in patients aged 40 ± 3 years (range 35-47 years) with an indication for polar body-based aneuploidy screening. Thirty-five aneuploid vitrified Day 3 embryos were warmed, cultured to Day 5 and biopsied for microarray analysis. Predictions were made for the ploidy status of the embryo if PSSC or non-disjunction had occurred. The signal intensity for the aneuploid chromosome in the first polar body was compared between those that resulted in euploid and aneuploid embryos. MAIN RESULTS AND THE ROLE OF CHANCE: Among 34 embryos with evaluable results, 31 were euploid on re-analysis. Of 43 chromosomes that had reciprocal aneuploidy in the polar bodies, 41 were disomic in the embryo, indicating that PSSC was likely to have occurred 95% (95% confidence interval 85-99%) of the time. The log 2 ratio signal intensity from the chromosomes that underwent non-disjunction, resulting in unbalanced embryos, were outliers when compared with those that underwent PSSC. LIMITATIONS, REASONS FOR CAUTION: Although most embryos with reciprocal aneuploid polar bodies were euploid, it is unknown whether they maintain equivalent reproductive potential when transferred. Further study is needed to determine whether these embryos should be re-biopsied and considered for transfer. WIDER IMPLICATIONS OF THE FINDINGS: This study is consistent with increasing evidence that PSSC is the primary cause of meiosis I errors in embryos from women of advanced reproductive age. Clinicians should be cautious in interpreting results from polar body aneuploidy screening, especially when only the first polar body is tested.
Subject(s)
Aneuploidy , Chromosome Aberrations , Embryo, Mammalian/physiology , Polar Bodies , Adult , Chromatids/metabolism , Chromatids/physiology , Cytogenetic Analysis , Female , Humans , Maternal Age , Meiosis , Middle Aged , Oligonucleotide Array Sequence Analysis , Polymorphism, Single Nucleotide , Preimplantation DiagnosisABSTRACT
BACKGROUND: The aim of this study was to examine the effect of the cryopreservation procedure (slow freezing or vitrification) and cryoprotectants (1,2-propanediol or dimethylsulphoxide) on mouse blastocyst gene expression. METHODS: Cultured mouse blastocysts were cryopreserved with different protocols. Following thawing/warming, total RNA from re-expanded blastocysts was isolated, amplified and then analyzed using mouse whole-genome microarrays. RESULTS: Compared with non-cryopresevered control blastocysts, gene expression was only significantly altered by slow freezing. Slow freezing affected the expression of 115 genes (P < 0.05). Of these, 100 genes exhibited down-regulation and 15 genes were up-regulated. Gene ontology revealed that the majority of these genes are involved in protein metabolism, transcription, cell organization, signal transduction, intracellular transport, macromolecule biosynthesis and development. Neither of the vitrification treatment groups showed statistically different gene expression from the non-cryopreserved control embryos. Hierarchical cluster analysis, did however, reveal that vitrification using 1,2-propanediol could result in a gene expression profile closest to that of non-cryopreserved blastocysts. CONCLUSIONS: Investigating the effects of cryopreservation on cellular biology, such as gene expression, is fundamental to improving techniques and protocols. This study demonstrates that of the cryopreservation regimens employed, slow freezing induced the most changes in gene expression compared with controls.
Subject(s)
Blastocyst/cytology , Cryopreservation/methods , Gene Expression Regulation , Animals , Cryoprotective Agents/pharmacology , Dimethyl Sulfoxide/pharmacology , Female , Freezing , Gene Expression Profiling , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Oligonucleotide Array Sequence Analysis , Propylene Glycol/pharmacology , Real-Time Polymerase Chain Reaction , VitrificationABSTRACT
Polycystic ovaries (PCO) is a common phenotype of women presenting for infertility treatment. This study investigated whether blastocysts derived from women with PCO have an altered molecular signature which could be a causative factor contributing to reproductive failure. Morphologically similar blastocysts derived from women with PCO and donor oocyte cycles were analysed for transcription and protein secretion. Unsupervised hierarchical clustering demonstrated that the transcriptome profiles of blastocysts derived from PCO patients and control blastocysts were markedly different with complete branch separation. Statistical analysis revealed 829 genes with significantly different expression: 784 decreased (94.6%) and 45 increased (5.4%) in blastocysts derived from women with PCO compared with controls (P<0.05). Functional annotation of these genes revealed predominant gene ontology biological processes including protein metabolism (30%), transcription (22%), signal transduction (15%), biosynthesis (15%) and cell cycle (14%). Proteomic profiling identified 12 biomarkers that displayed significant decrease in expression in blastocysts derived from women with PCO compared with controls (P<0.05). These data indicate molecular alterations in human blastocysts derived from PCO patients, potentially demonstrating for the first time a link between patient aetiology/phenotype and subsequent embryo development, which in part may explain the observed reduction in reproductive capacity.
Subject(s)
Blastocyst/metabolism , Polycystic Ovary Syndrome/metabolism , Proteome/analysis , Female , Gene Expression Profiling , HumansABSTRACT
Previous studies have shown that electrical charges influence cell behavior (e.g. enhancement of nerve regeneration, cell adhesion, cell morphology). Thus, piezoelectric scaffolds might be useful for various tissue engineering applications. Fibrous scaffolds were successfully fabricated from permanent piezoelectric poly(vinylidene fluoride-trifluoroethylene) (PVDF-TrFE) by the electrospinning technique. Scanning electron microscopy and capillary flow analyses verified that the fiber mats had an average fiber diameter of 970 +/- 480 nm and a mean pore diameter of 1.7 microm, respectively. Thermally stimulated depolarization current spectroscopy measurements confirmed the piezoelectric property of the PVDF-TrFE fibrous scaffolds by the generation of a spontaneous current with the increase in temperature in the absence of an electric field, which was not detected in the unprocessed PVDF-TrFE powder. Differential scanning calorimetry, thermogravimetric analysis, X-ray diffraction and Fourier transform infrared spectroscopy results showed that the electrospinning process increased the crystallinity and presence of the polar, beta-phase crystal compared with the unprocessed powder. Confocal fluorescence microscopy and a cell proliferation assay demonstrated spreading and increased cell numbers (human skin fibroblasts) over time on PVDF-TrFE scaffolds, which was comparable with tissue culture polystyrene. The relative quantity of gene expression for focal adhesion proteins (measured by real-time RT-PCR) increased in the following order: paxillin < vinculin < focal adhesion kinase < talin. However, no differences could be seen among the TCPS surface and the fibrous scaffolds. Future studies will focus on possible applications of these cytocompatible PVDF-TrFE scaffolds in the field of regenerative medicine.
Subject(s)
Biocompatible Materials/pharmacology , Electricity , Fibroblasts/cytology , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Calorimetry, Differential Scanning , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Fibroblasts/drug effects , Fibroblasts/metabolism , Gene Expression Regulation/drug effects , Humans , Microscopy, Confocal , Microscopy, Electron, Scanning , Polyvinyls/chemistry , Porosity/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Spectroscopy, Fourier Transform Infrared , Temperature , X-Ray DiffractionABSTRACT
Polyarylates have shown promise as fully degradable polymers for drug delivery as well as for structural implant applications due to their range of physicomechanical properties. Processing history, however, could have a significant impact on their overall performance in biologically relevant environments. More specifically, structural changes at the molecular level can occur that will affect a polymer's physical properties and subsequent, cell attachment and growth. The present study was aimed at comparing cell growth on tyrosine-derived polyarylates with that of polylactic acid (PLLA) in their original state and after processing (i.e. undrawn and drawn forms). Two polyarylates having distinct molecular structures were chosen. Strictly, amorphous poly(DTE adipate), denoted as poly(DT 2,4), and poly(DTD) dodecandioate, denoted as poly(DT 12,10), having a more complex, non-crystalline organization, were compared with semi-crystalline PLLA. The degree of shrinkage, thermal characterization, air-water contact angle and surface morphology were determined for each polymer in its undrawn and drawn states. Poly(DT 2,4) and PLLA after processing resulted in greater shrinkage and a slight decrease in hydrophilicity whereas poly(DT 12,10) had minimal shrinkage and became slightly more hydrophilic in its drawn state. Surface morphology or roughness was also altered by processing. In turn, the rate of cell growth and overall cell numbers were reduced significantly on drawn forms of poly(DT 2,4) and PLLA, whereas more favorable growth rates were supported on drawn poly(DT 12,10). These findings indicate that processing effects in amorphous as well as oriented polymeric structures can significantly alter their biological performance.
Subject(s)
Lactic Acid/chemistry , Polyesters/chemistry , Polymers/chemistry , Tyrosine/chemistry , Air , Calorimetry, Differential Scanning/methods , Crystallization , Fibroblasts/metabolism , Humans , Microscopy, Electron, Scanning/methods , Models, Chemical , Surface Properties , Thermogravimetry/methods , Time Factors , Water/chemistryABSTRACT
Non-invasive gamete and embryo assessment is considered an important focus in assisted reproductive technologies (ART). Currently, the selection of embryos for transfer is based on morphological indices. Though successful, the field of ART would benefit from a non-invasive quantitative method of viability determination. Omics technologies, including transcriptomics, proteomics and metabolomics, have already begun providing evidence that viable gametes and embryos possess unique molecular profiles with potential biomarkers that can be utilized for developmental and/or viability selection. Unlike the human genome that is relatively fixed and steady throughout the human body, the human proteome, estimated at over a million proteins, is more complex, diverse and dynamic. It is the proteins themselves that contribute to the physiological homeostasis in any cell or tissue. Of particular interest in ART is the secretome, those proteins that are produced within the embryo and secreted into the surrounding environment. Defining the human embryonic secretome has the potential to expand our knowledge of embryonic cellular processes, including the complex dialogue between the developing embryo and its maternal environment, and may also assist in identifying those embryos with the highest implantation potential. Advances in proteomic technologies have allowed the non-invasive profiling of the human embryonic secretome with ongoing research focused on correlation with outcome. From a clinical perspective, embryo selection based on morphological assessment and non-invasive analysis of the human embryonic secretome may improve IVF success and lead to routine single embryo transfers.
Subject(s)
Embryo, Mammalian/metabolism , Metabolomics , Proteome , Proteomics , Animals , Biomarkers/metabolism , Humans , Mass Spectrometry , Preimplantation Diagnosis/methods , Protein Array Analysis , Proteome/analysis , Proteome/metabolism , Reproductive Techniques, AssistedABSTRACT
In vitro maturation (IVM) of mammalian oocytes does not support the same rates of embryo development or pregnancy when compared to oocytes that have matured in vivo. Therefore, environment has a significant influence on the oocyte's ability to complete maturation and acquire the mRNA and proteins required for successful fertilization and normal embryonic development. The aim of this study was to analyze the MII oocyte transcriptome between in vivo and in vitro conditions. Total RNA was extracted, processed and hybridized to the Affymetrix GeneChip Bovine Genome Array. Following normalization of the microarray data, analysis revealed 10 differentially expressed genes after IVM compared to in vivo matured controls, including Aqp3, Sept7, Abhd4 and Siah2 (P<0.05). K-means cluster analysis coupled with associated gene ontology, identified several biological processes affected by IVM, including metabolism, energy pathways, cell organization and biogenesis, and cell growth and maintenance. Quantitative real-time PCR validated the microarray data and also revealed altered expression levels after IVM of specific putatively imprinted genes, Igf2r, Peg3 and Snrpn (P<0.05). Distinct IVM transcription patterns reflected the oocyte's response to its surrounding environment. Monitoring transcription levels of key oocyte maturation genes may subsequently assist in improving IVM success.
Subject(s)
Cattle , Gene Expression Profiling/veterinary , Oocytes/chemistry , Oocytes/growth & development , Animals , Carbohydrate Metabolism , Energy Metabolism , Female , In Vitro Techniques , Oligonucleotide Array Sequence Analysis/veterinary , Oocytes/metabolism , Polymerase Chain Reaction/veterinaryABSTRACT
Proteomics describes the changes in all proteins expressed and translated from a single genome. At present little is known regarding either the genome or proteome of human gametes or the preimplantation embryo. The unravelling of this information is fundamental to understanding the complexity of reproductive physiology, including the dialogue between the developing embryo and its maternal environment. To date, a lack of sensitivity has been the main reason behind the inability to introduce proteomics technology into assisted reproduction techniques. Proteomics alone involves several sophisticated techniques including imaging, mass spectrometry and bioinformatics to identify, quantify and characterize a proteome. The recent increased sensitivity of these techniques has allowed for the development of new protocols that are capable of not only profiling the proteome of individual human oocytes and embryos, but also the proteins produced by the embryo into the surrounding medium (the secretome). Hence, the identification of proteins that are involved in oocyte maturation, embryo development and implantation could lead to further improvements in assisted reproduction techniques as well as the development of new diagnostic tests. Furthermore, proteomics may contribute in the design of a non-invasive viability assay to assist in the selection of embryos for transfer in human assisted reproduction.