RESUMO
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.
Assuntos
Diagnóstico Pré-Implantação , Aneuploidia , Blastocisto , Transferência Embrionária , Feminino , Testes Genéticos/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Mosaicismo , Gravidez , Diagnóstico Pré-Implantação/métodosRESUMO
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.
Assuntos
Fertilidade/genética , Obesidade/genética , Proteoma/genética , Espermatogênese/genética , Adulto , Índice de Massa Corporal , Feminino , Humanos , Infertilidade Masculina/complicações , Infertilidade Masculina/genética , Infertilidade Masculina/metabolismo , Infertilidade Masculina/patologia , Masculino , Obesidade/complicações , Obesidade/patologia , Estresse Oxidativo/genética , Proteoma/metabolismo , Análise do Sêmen , Contagem de Espermatozoides , Motilidade dos Espermatozoides/genética , Espermatozoides/metabolismo , Espermatozoides/patologiaRESUMO
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.
Assuntos
Respiração , Fenômenos Biomecânicos , Dióxido de Carbono/metabolismo , Impedância Elétrica , Humanos , Intubação Intratraqueal , Pulmão/diagnóstico por imagem , Pulmão/fisiologia , Monitorização Fisiológica , Pressão , Troca Gasosa Pulmonar , Ventilação Pulmonar , Respiração Artificial , Taxa Respiratória , TomografiaRESUMO
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).
Assuntos
Neuropatias Diabéticas/terapia , Monitorização Fisiológica/métodos , Atenção Primária à Saúde , Qualidade de Vida , Idoso , Análise por Conglomerados , Neuropatias Diabéticas/fisiopatologia , Neuropatias Diabéticas/psicologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Padrões de Prática MédicaRESUMO
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.
Assuntos
Inversão Cromossômica/genética , Desenvolvimento Embrionário/genética , Fertilização in vitro , Infertilidade/genética , Adulto , Aneuploidia , Blastocisto/patologia , Hibridização Genômica Comparativa , Transferência Embrionária , Feminino , Testes Genéticos , Humanos , Infertilidade/fisiopatologia , Idade Materna , Gravidez , Taxa de Gravidez , Diagnóstico Pré-ImplantaçãoRESUMO
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.
Assuntos
Monitorização Fisiológica/métodos , Publicações Periódicas como Assunto , Respiração , Taxa Respiratória , Animais , Capnografia , Dióxido de Carbono/química , Ensaios Clínicos como Assunto , Impedância Elétrica , Humanos , Pulmão/fisiologia , Monitorização Fisiológica/instrumentação , Oximetria , Troca Gasosa Pulmonar , Respiração Artificial , Processamento de Sinais Assistido por ComputadorRESUMO
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.
Assuntos
Monitorização Fisiológica/instrumentação , Monitorização Fisiológica/métodos , Oximetria/métodos , Respiração , Taxa Respiratória , Algoritmos , Animais , Capnografia , Dióxido de Carbono , Impedância Elétrica , Eletrocardiografia , Humanos , Período Pós-Operatório , Troca Gasosa Pulmonar , Respiração ArtificialRESUMO
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.
Assuntos
Pneumopatias/diagnóstico , Publicações Periódicas como Assunto/tendências , Polissonografia/tendências , Transtornos Respiratórios/diagnóstico , Testes de Função Respiratória/tendências , Humanos , Pneumopatias/prevenção & controle , Polissonografia/instrumentação , Polissonografia/métodos , Transtornos Respiratórios/prevenção & controle , Testes de Função Respiratória/instrumentação , Testes de Função Respiratória/métodosAssuntos
Quimera/embriologia , Transferência Embrionária/normas , Mosaicismo , Diagnóstico Pré-Implantação/normas , Transferência Embrionária/métodos , Feminino , Humanos , Internacionalidade , Gravidez , Resultado da Gravidez/genética , Diagnóstico Pré-Implantação/métodos , Técnicas de Reprodução Assistida/normas , Sociedades Médicas/normasRESUMO
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.
Assuntos
Aneuploidia , Aberrações Cromossômicas , Embrião de Mamíferos/fisiologia , Corpos Polares , Adulto , Cromátides/metabolismo , Cromátides/fisiologia , Análise Citogenética , Feminino , Humanos , Idade Materna , Meiose , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Diagnóstico Pré-ImplantaçãoRESUMO
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.
Assuntos
Compostos de Amônio/efeitos adversos , Técnicas de Cultura Embrionária , Embrião de Mamíferos/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Animais , Embrião de Mamíferos/metabolismo , Feminino , Humanos , Metabolismo/efeitos dos fármacos , Camundongos , GravidezRESUMO
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.
Assuntos
Blastocisto/citologia , Criopreservação/métodos , Regulação da Expressão Gênica , Animais , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Feminino , Congelamento , Perfilação da Expressão Gênica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Análise de Sequência com Séries de Oligonucleotídeos , Propilenoglicol/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , VitrificaçãoRESUMO
The smaller form of the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD65) is a major autoantigen in two human diseases that affect its principal sites of expression. Thus, destruction of pancreatic beta cells, which results in insulin-dependent diabetes mellitus (IDDM), and impairment of GABA-ergic synaptic transmission in Stiff-Man syndrome (SMS) are both characterized by circulating autoantibodies to GAD65. Anti-GAD65 autoantibodies in IDDM are predominantly directed to conformational epitopes. Here we report the characterization of humoral autoimmune responses to GAD65 in 35 SMS patients, of whom 13 (37%) also had IDDM. All SMS patients immunoprecipitated native GAD65 and the main titers were orders of magnitude higher than in IDDM patients. Furthermore, in contrast to the situation in IDDM, autoantibodies in 35 of 35 (100%) of SMS patients recognized denatured GAD65 on Western blots. Two major patterns of epitope specificity were identified on Western blots. The first pattern, detected in 25 of 35 SMS patients (71%), of whom 11 had IDDM (44%), was predominantly reactive with a linear NH2-terminal epitope residing in the first eight amino acids of GAD65. Nine of nine individuals who were HLA-haplotyped in this group carried an IDDM susceptibility haplotype and HLA-DR3, DQw2 was particularly abundant. The second pattern, detected in 10 of 35 patients (29%) of whom two had IDDM (20%), included reactivity with the NH2-terminal epitope plus strong reactivity with one or more additional epitope(s) residing COOH-terminal to amino acid 101. The second epitope pattern may represent epitope spreading in the GAD65 molecule, but may also include some cases of epitope recognition associated with IDDM resistant HLA-haplotypes. The principal NH2-terminal linear epitope in GAD65 distinguishes the reactivity of SMS and IDDM autoantibodies and may be a determinant of pathogenicity for GABA-ergic neurons. The greater magnitude and distinct specificity of the humoral response to GAD65 in SMS may reflect a biased involvement of the T helper cell type 2 (Th2) subset of CD4+ T cells and antibody responses, whereas IDDM is likely mediated by the Th1 subset of CD4+ T cells and cytotoxic T cell responses.
Assuntos
Autoanticorpos/imunologia , Autoantígenos/imunologia , Diabetes Mellitus Tipo 1/imunologia , Epitopos/imunologia , Glutamato Descarboxilase/imunologia , Rigidez Muscular Espasmódica/imunologia , Adulto , Idoso , Especificidade de Anticorpos , Feminino , Antígenos HLA-DQ/genética , Cadeias beta de HLA-DQ , Antígenos HLA-DR/genética , Cadeias HLA-DRB1 , Haplótipos , Antígenos de Histocompatibilidade Classe II/genética , Humanos , Masculino , Pessoa de Meia-Idade , Testes de PrecipitinaRESUMO
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.
Assuntos
Blastocisto/metabolismo , Síndrome do Ovário Policístico/metabolismo , Proteoma/análise , Feminino , Perfilação da Expressão Gênica , HumanosRESUMO
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.
Assuntos
Embrião de Mamíferos/metabolismo , Metabolômica , Proteoma , Proteômica , Animais , Biomarcadores/metabolismo , Humanos , Espectrometria de Massas , Diagnóstico Pré-Implantação/métodos , Análise Serial de Proteínas , Proteoma/análise , Proteoma/metabolismo , Técnicas de Reprodução AssistidaRESUMO
When illuminated pea tendrils from light-grown plants are rubbed on their abaxial side, they rapidly coil in a spiral fashion. If similar tendrils are held in the dark for 3 days and then rubbed, however, they will not coil until they are subsequently illuminated. They can remain uncoiled in the dark for as long as 2 hours after stimulation, and will still coil immediately when they are illuminated. Tendrils that are rubbed and held at 25 degrees C will coil, but those treated at 5 degrees or 10 degrees C will not. However, tendrils rubbed at 25 degrees C and kept from coiling for an hour at 5 degrees C, will immediately coil when restored to the higher temperature. These observations are interpreted to imply separation of sensory and motor functions.
RESUMO
Excised root tips from dark-grown mung bean seedlings (Phaseolus aureus) that adhere to a negatively charged glass surface when irradiated with red light and release when irradiated by far-red light develop a positive bioelectric potential (about 1.0 millivolt) at the tip in red light and a negative bioelectric potential in far-red light. The sign of the bioelectric potential was repeatedly photo-reversible, and the adhesion and release kinetics were similar to those of the development of the bioelectric potentials. Photoconversion of the phytochrome holochrome perhaps changes permeability characteristics of the cell membrane, resulting in an induced localized electrochemical gradient manifested as a bioelectric potential. This supports the view that phytochrome in situ is membrane bound.
Assuntos
Potenciais Evocados , Luz , Pigmentos Biológicos/fisiologia , Reguladores de Crescimento de Plantas/fisiologia , Proteínas de Plantas/fisiologia , Plantas Comestíveis/fisiologia , Cinética , Plantas Comestíveis/efeitos da radiação , Efeitos da Radiação , SementesRESUMO
Plant roots grow in the direction of increasing soil moisture, but studies of hydrotropism have always been difficult to interpret because of the effect of gravity. In this study it was found that roots of the mutant pea ;Ageotropum' are neither gravitropic nor phototropic, but do respond tropically to a moisture gradient, making them an ideal subject for the study of hydrotropism. When the root caps were removed, elongation was not affected but hydrotropism was blocked, suggesting that the site of sensory perception resides in the root cap.
RESUMO
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.