DOCSLIB.ORG

Reproduction

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Reproduction REPRODUCTIONRESEARCH Focus on Mammalian Embryogenomics Molecular and subcellular characterisation of oocytes screened for their developmental competence based on glucose-6-phosphate dehydrogenase activity Helmut Torner2, Nasser Ghanem, Christina Ambros2, Michael Ho¨lker, Wolfgang Tomek2, Chirawath Phatsara, Hannelore Alm2, Marc-Andre´ Sirard1, Wilhelm Kanitz2, Karl Schellander and Dawit Tesfaye Animal Breeding and Husbandry Group, Department of Animal Breeding and Husbandry, Institute of Animal Science, University of Bonn, Endenicher allee 15, 53115 Bonn, Germany, 1De´partement des Sciences Animales, Centre de Recherche en Biologie de la Reproduction, Universite´ Laval, Pav. Comtois, Laval, Sainte-Foy, Que´bec, G1K 7P4, Canada and 2Research Institute for the Biology of Farm Animals, Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany Correspondence should be addressed to D Tesfaye; Email: [email protected] Abstract Oocyte selection based on glucose-6-phosphate dehydrogenase (G6PDH) activity has been successfully used to differentiate between competent and incompetent bovine oocytes. However, the intrinsic molecular and subcellular characteristics of these oocytes have not yet been investigated. Here, we aim to identify molecular and functional markers associated with oocyte developmental potential when selected based on G6PDH activity. Immature compact cumulus–oocyte complexes were stained with brilliant cresyl blue (BCB) for 90 min. Based on K C their colouration, oocytes were divided into BCB (colourless cytoplasm, high G6PDH activity) and BCB (coloured cytoplasm, low G6PDH activity). The chromatin configuration of the nucleus and the mitochondrial activityof oocytes were determined by fluorescence labelling and photometric measurement. The abundance and phosphorylation pattern of protein kinases Akt and MAP were estimated by Western blot C K analysis. A bovine cDNA microarray was used to analyse the gene expression profiles of BCB and BCB oocytes. Consequently, marked C K C differences were found in blastocyst rate at day 8 between BCB (33.1G3.1%) and BCB (12.1G1.5%) oocytes. Moreover, BCB oocytes were found to show higher phosphorylation levels of Akt and MAP kinases and are enriched with genes regulating transcription (SMARCA5), K cell cycle (nuclear autoantigenic sperm protein, NASP ) and protein biosynthesis (RPS274A and mRNA for elongation factor 1a, EF1A). BCB oocytes, which revealed higher mitochondrial activity and still nucleoli in their germinal vesicles, were enriched with genes involved in ATP synthesis (ATP5A1), mitochondrial electron transport (FL405), calcium ion binding (S100A10) and growth factor activity (bone morphogenetic protein 15, BMP15). This study has evidenced molecular and subcellular organisational differences of oocytes with different G6PDH activity. Reproduction (2008) 135 197–212 Introduction dairy cows leading to higher economic loss (Macmillan et al. 1996). This decline in fertility can be explained by In modern animal agriculture, with increasing milk management changes within the dairy industry and also production there is a continuous decline in the fertility of negative genetic correlations between milk production and reproduction. One of the primary mechanisms that This article was presented at the 2nd International Meeting on depresses fertility in lactating cows is abnormal pre- Mammalian Embryogenomics, 17–20 October 2007. The Co-operative implantation embryo development, which that may be a Research Programme: Biological Resource Management for Sustain- result of poor oocyte quality (Snijders et al. 2000, Lucy able Agricultural Systems of The Organisation for Economic Co- 2007). Oocyte developmental competence is defined operation and Development (OECD) has supported the publication of as the ability of an oocyte to resume meiosis, to cleave this article. The meeting was also sponsored by Le conseil Re´gional Ile- de-France, the Institut National de la Recherche Agronomique (INRA), following fertilisation, to develop to the blastocyst stage, Cogenics-Genome Express, Eurogentec, Proteigene, Sigma-Aldrich to induce a pregnancy and bring offspring to term in a France and Diagenode sa. good health (Krisher 2004, Sirard et al. 2006). This q 2008 Society for Reproduction and Fertility DOI: 10.1530/REP-07-0348 ISSN 1470–1626 (paper) 1741–7899 (online) Online version via www.reproduction-online.org Downloaded from Bioscientifica.com at 09/30/2021 08:04:27AM via free access 198 H Torner and others competency is acquired gradually during the course of molecular and the subcellular characteristics of these folliculogenesis as the oocyte grows and its companion oocytes. Therefore, the aim of this study was to somatic cells differentiate (Eppig et al. 1994). characterise these oocytes at the subcellular level Many factors have been shown to affect the oocyte’s (dissolution of nucleoli and mitochondrial activity), developmental potential, including follicle size molecular level (gene expression profile) and function- (Lonergan et al. 1994), health of the follicle (Blondin & ally (activity of protein kinase). The results of the present Sirard 1995, Vassena et al. 2003), phase of follicular study evidence the prevailing differences of these wave (Hagemann 1999, Machatkova´ et al.2004), oocyte groups in relative abundance transcripts and hormonal stimulation (Blondin et al. 2002; for review mitochondrial and MAPK activities contributing to their Sirard et al. 2006), maturation environment (Warzych differences in developmental potential. et al. 2007; for review Sutton et al. 2003), season (Al-Katanani et al. 2002, Sartori et al. 2002), nutrition (Fouladi-Nashta et al. 2007) and age (Rizos et al. 2005). Although previous studies support the notion that oocyte Results competence depends on multiple factors, it remains difficult Chromatin configuration and mitochondrial activity in C K to draw clear and reliable criteria for oocyte selection. BCB and BCB oocytes Morphological assessment of oocytes based on thickness, compactness of the cumulus investment and Because of their importance as parameters for oocyte the homogeneity of the ooplasm (Gordon 2003)isa quality, we investigated the status of nuclei and mito- C K relatively popular and convenient way of evaluating chondria in BCB and BCB oocytes before in vitro oocyte quality in practice. However, results derived from maturation (IVM). A larger proportion of oocytes with K this non-invasive approach are often conflicting, largely high G6PDH activity (BCB ) were found to be in early due to subjectivity and inaccuracy. Morphological stage of diplotene with clear visible nucleoli (DiplCNuc) C evaluation alone is insufficient to distinguish competent in their germinal vesicle than the BCB oocytes (Table 1; oocytes that have the ability to bring about full-term P!0.005). However, a significantly lower number of a K pregnancy (Lonergan et al. 2003, Coticchio et al. 2004, BCB oocytes was found to be in more progressed Krisher 2004). With the urgent need for establishing non- diakinesis stage after germinal vesicle breakdown C invasive and non-perturbing means for oocyte selection, (GVBD) compared with their BCB counterparts. the brilliant cresyl blue (BCB) staining test has been To confirm that the fluorescence intensity of the successfully used to differentiate oocytes with different emission light from the fixed MitoTracker-labelled oocytes developmental capacity in various species, including pig was stable during the time of storage, a preliminary study (Ericsson et al. 1993, Roca et al.1998, Wongsrikeao et al. was conducted to measure the fluorescence intensity of 40 2006), goat (Rodrı´guez-Gonza´lez et al. 2002) and cattle oocytes in intervals of 7 days during 6 weeks. The (Alm et al. 2005, Bhojwani et al. 2007). measured fluorescence intensity was not influenced by During the course of their growth, immature oocytes the storage. are known to synthesise a variety of proteins, including The data in Table 1 demonstrate that the fluorescence glucose-6-phosphate dehydrogenase (G6PDH; Mangia intensity in the oocytes pre-labelled by the vital & Epstein 1975, Wassarman 1988). The activity of this mitochondrial-specific probe chloromethyl tetramethyl- protein is decreased once this phase has been completed rosamine (CMTM Ros) and measured by fluorescence and oocytes are then likely to have achieved develop- intensity for 570 nm emission/oocyte is associated with mental competence (Wassarman 1988, Tian et al. 1998). their G6PDH activity (P!0.001). The highest fluor- BCB is a dye that can be degraded by G6PDH (Ericsson K escence intensity/oocyte was found in BCB oocytes et al. 1993, Tian et al. 1998); thus, oocytes that have C finished their growth phase show decreased G6PDH compared with the BCB ones. activity and exhibit cytoplasm with a blue colouration C (BCB ), while growing oocytes are expected to have a Detection of abundance and phosphorylation of protein high level of active G6PDH, which results in colourless K kinases Akt and MAP cytoplasm (BCB ). In our previous studies, it has been shown that In order to elucidate the activities of protein kinases that oocytes screened based on BCB staining differ in their contribute in the regulation of gene expression, we have developmental potential to reach blastocyst stage (Alm analysed the abundance and phosphorylation state of et al. 2005) and efficiency in utilisation for somatic cell the MAPKs ERK1, ERK2 and Akt. As indicated in Fig. 1, the abundance of MAPK and Akt1 was not different nuclear transfer (Bhojwani et al.
Load more

Recommended publications
  • Comprehensive Identification and Characterization of Somatic Copy Number Alterations in Triple‑Negative Breast Cancer
    INTERNATIONAL JOURNAL OF ONCOLOGY 56: 522-530, 2020 Comprehensive identification and characterization of somatic copy number alterations in triple‑negative breast cancer ZAIBING LI1,2*, XIAO ZHANG3*, CHENXIN HOU4, YUQING ZHOU4, JUNLI CHEN1, HAOYANG CAI5, YIFENG YE3, JINPING LIU3 and NING HUANG1 1Department of Pathophysiology, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041; 2Department of Pathophysiology, School of Basic Medical Science, Southwest Medical University, Luzhou, Sichuan 646000; 3Department of Breast Surgery, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan 611731; 4West China Medical School, Sichuan University, Chengdu, Sichuan 610041; 5Center of Growth, Metabolism and Aging, Key Laboratory of Bio‑Resources and Eco‑Environment, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610064, P.R. China Received January 30, 2019; Accepted August 30, 2019 DOI: 10.3892/ijo.2019.4950 Abstract. Triple-negative breast cancer (TNBC) accounts hierarchical clustering of tumors resulted in three main for ~15% of all breast cancer diagnoses each year. Patients subgroups that exhibited distinct CNA profiles, which with TNBC tend to have a higher risk for early relapse and may reveal the heterogeneity of molecular mechanisms in a worse prognosis. TNBC is characterized by extensive TNBC subgroups. These results will extend the molecular somatic copy number alterations (CNAs). However, the DNA understanding of TNBC and will facilitate the discovery of CNA profile of TNBC remains to be extensively investigated. therapeutic and diagnostic target candidates. The present study assessed the genomic profile of CNAs in 201 TNBC samples, aiming to identify recurrent CNAs that Introduction may drive the pathogenesis of TNBC.
    [Show full text]
  • Differentially Expressed on Collagen Networks 1, 2, 10 © 2000-2009 Ingenuity Systems, Inc. All Rights Reserved. Symbol Entrez
    Differentially expressed on collagen Networks 1, 2, 10 © 2000-2009 Ingenuity Systems, Inc. All rights reserved. Symbol Entrez Gene Name Affymetrix Fold Change Location Family ALDH1A3 aldehyde dehydrogenase 1 family, member A3 203180_at -5.43125168 Cytoplasm enzyme 209772_s_ Plasma CD24 CD24 molecule at -4.32890229 Membrane other HSD11B2 hydroxysteroid (11-beta) dehydrogenase 2 204130_at -4.1099197 Cytoplasm enzyme Plasma AMOTL2 angiomotin like 2 203002_at -2.82872773 Membrane other transcription DLX2 distal-less homeobox 2 207147_at -2.74996362 Nucleus regulator 221215_s_ RIPK4 receptor-interacting serine-threonine kinase 4 at -2.56556472 Nucleus kinase PLK2 polo-like kinase 2 (Drosophila) 201939_at -2.47054478 Nucleus kinase ALDH3A1 aldehyde dehydrogenase 3 family, memberA1 205623_at -2.30532989 Cytoplasm enzyme TXNRD1 thioredoxin reductase 1 201266_at -2.27936909 Cytoplasm enzyme Extracellular CYR61 cysteine-rich, angiogenic inducer, 61 201289_at -2.09052668 Space other 214212_x_ FERMT2 fermitin family homolog 2 (Drosophila) at -1.87478183 Cytoplasm other Plasma RIT1 Ras-like without CAAX 1 209882_at -1.77586775 Membrane enzyme 210297_s_ Extracellular MSMB microseminoprotein, beta- at -1.72177723 Space other Extracellular PI3 peptidase inhibitor 3, skin-derived 203691_at -1.68135697 Space other ALDH3B1 aldehyde dehydrogenase 3 family, member B1 205640_at -1.67376791 Cytoplasm enzyme 202124_s_ Plasma TRAK2 trafficking protein, kinesin binding 2 at -1.6367793 Membrane transporter BMP and activin membrane-bound inhibitor Plasma BAMBI
    [Show full text]
  • MOLECULAR G EN ETICS of the January, HUMAN X
    MOLECULAR GENETICS OF THE HUMAN X CHROMOSOME BY DAVID ANDREW HARTLEY a thesis submitted for the degree of Doctor of Philosophy in the University of London January, 1984 Department of Biochemistry St. Mary's Hospital Medical School London W2 IPG 1 Abstract The comparison of frequency of recombination with chromosomal position has been possible only in Drosophila melanogaster but it is known that chiasma distribution in the human male (and many other species) is not random. A number of cloned DNA sequences have been isolated from a library enriched for X chromosome DNA by flow cytometry. The chromosomal loci complementary to these cloned sequences have been investigated by hybridization to somatic cell hybrid DNAs and by direct hybridization 1 in situ' to human metaphase spreads. In this way a cytological map of the X chromosome defined by DNA sequence markers has been constructed. Genetic distances complementary to the physical separations determined have been estimated by determining the segregation of DNA sequence variants at each locus in three generation p e d i g r e e s . The sequence variation is manifested as altered restriction fragment migration in agarose gels. It is intimated from these analyses that the pattern of recombination frequency along the human X chromosome mirrors the non-random chiasma distribution seen along male autosomes. This supports the chiasmatype theory and has implications on the availability of genetic components of the X chromosome to recombination. 2 To ray father in the hope of convincing him of life outside o f E.coli 3 ...scientists getting their kicks when deadly disease can do as it pleases results ain't hard to predict Gil Scott-Heron 4 Ack n ov; ledgements To my supervisor Bob Williamson for support.
    [Show full text]
  • 1471-2164-10-261.Pdf
    BMC Genomics BioMed Central Research article Open Access Molecular signature of cell cycle exit induced in human T lymphoblasts by IL-2 withdrawal Magdalena Chechlinska*1, Jan Konrad Siwicki1, Monika Gos2, Malgorzata Oczko-Wojciechowska3, Michal Jarzab4, Aleksandra Pfeifer3, Barbara Jarzab3 and Jan Steffen1 Address: 1Department of Immunology, Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Warsaw, Poland, 2Department of Cell Biology, Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Warsaw, Poland, 3Department of Nuclear Medicine and Endocrine Oncology, Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Gliwice, Poland and 4Department of Tumor Biology and Clinical Oncology, Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Gliwice, Poland Email: Magdalena Chechlinska* - [email protected]; Jan Konrad Siwicki - [email protected]; Monika Gos - [email protected]; Malgorzata Oczko-Wojciechowska - [email protected]; Michal Jarzab - [email protected]; Aleksandra Pfeifer - [email protected]; Barbara Jarzab - [email protected]; Jan Steffen - [email protected] * Corresponding author Published: 8 June 2009 Received: 20 February 2009 Accepted: 8 June 2009 BMC Genomics 2009, 10:261 doi:10.1186/1471-2164-10-261 This article is available from: http://www.biomedcentral.com/1471-2164/10/261 © 2009 Chechlinska et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: The molecular mechanisms of cell cycle exit are poorly understood.
    [Show full text]
  • Combined Analysis of Genome-Wide Expression and Copy Number
    Fontanillo et al. BMC Genomics 2012, 13(Suppl 5):S5 http://www.biomedcentral.com/1471-2164/13/S5/S5 RESEARCH Open Access Combined analysis of genome-wide expression and copy number profiles to identify key altered genomic regions in cancer Celia Fontanillo1, Sara Aibar1, Jose Manuel Sanchez-Santos2, Javier De Las Rivas1* From X-meeting 2011 - International Conference on the Brazilian Association for Bioinformatics and Compu- tational Biology Florianópolis, Brazil. 12-15 October 2011 Abstract Background: Analysis of DNA copy number alterations and gene expression changes in human samples have been used to find potential target genes in complex diseases. Recent studies have combined these two types of data using different strategies, but focusing on finding gene-based relationships. However, it has been proposed that these data can be used to identify key genomic regions, which may enclose causal genes under the assumption that disease-associated gene expression changes are caused by genomic alterations. Results: Following this proposal, we undertake a new integrative analysis of genome-wide expression and copy number datasets. The analysis is based on the combined location of both types of signals along the genome. Our approach takes into account the genomic location in the copy number (CN) analysis and also in the gene expression (GE) analysis. To achieve this we apply a segmentation algorithm to both types of data using paired samples. Then, we perform a correlation analysis and a frequency analysis of the gene loci in the segmented CN regions and the segmented GE regions; selecting in both cases the statistically significant loci. In this way, we find CN alterations that show strong correspondence with GE changes.
    [Show full text]
  • Download Special Issue
    International Journal of Genomics Noncoding RNAs in Health and Disease Lead Guest Editor: Michele Purrello Guest Editors: Massimo Romani and Davide Barbagallo Noncoding RNAs in Health and Disease International Journal of Genomics Noncoding RNAs in Health and Disease Lead Guest Editor: Michele Purrello Guest Editors: Massimo Romani and Davide Barbagallo Copyright © 2018 Hindawi. All rights reserved. This is a special issue published in “International Journal of Genomics.” All articles are open access articles distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Editorial Board Andrea C. Belin, Sweden M. Hadzopoulou-Cladaras, Greece Ferenc Olasz, Hungary Jacques Camonis, France Sylvia Hagemann, Austria Elena Pasyukova, Russia Prabhakara V. Choudary, USA Henry Heng, USA Graziano Pesole, Italy Martine A. Collart, Switzerland Eivind Hovig, Norway Giulia Piaggio, Italy Monika Dmitrzak-Weglarz, Poland Hieronim Jakubowski, USA Mohamed Salem, USA Marco Gerdol, Italy B.-H. Jeong, Republic of Korea Wilfred van IJcken, Netherlands João Paulo Gomes, Portugal Atsushi Kurabayashi, Japan Brian Wigdahl, USA Soraya E. Gutierrez, Chile Giuliana Napolitano, Italy Jinfa Zhang, USA Contents Noncoding RNAs in Health and Disease Davide Barbagallo, Gaetano Vittone, Massimo Romani , and Michele Purrello Volume 2018, Article ID 9135073, 2 pages Circular RNAs: Biogenesis, Function, and a Role as Possible Cancer Biomarkers Luka Bolha,
    [Show full text]
  • Candidate Genes for Alcohol Preference Identified by Expression
    Liang et al. Genome Biology 2010, 11:R11 http://genomebiology.com/2010/11/2/R11 RESEARCH Open Access Candidate genes for alcohol preference identified by expression profiling in alcohol-preferring and -nonpreferring reciprocal congenic rats Tiebing Liang1*, Mark W Kimpel2, Jeanette N McClintick3, Ashley R Skillman1, Kevin McCall4, Howard J Edenberg3, Lucinda G Carr1 Abstract Background: Selectively bred alcohol-preferring (P) and alcohol-nonpreferring (NP) rats differ greatly in alcohol preference, in part due to a highly significant quantitative trait locus (QTL) on chromosome 4. Alcohol consumption scores of reciprocal chromosome 4 congenic strains NP.P and P.NP correlated with the introgressed interval. The goal of this study was to identify candidate genes that may influence alcohol consumption by comparing gene expression in five brain regions of alcohol-naïve inbred alcohol-preferring and P.NP congenic rats: amygdala, nucleus accumbens, hippocampus, caudate putamen, and frontal cortex. Results: Within the QTL region, 104 cis-regulated probe sets were differentially expressed in more than one region, and an additional 53 were differentially expressed in a single region. Fewer trans-regulated probe sets were detected, and most differed in only one region. Analysis of the average expression values across the 5 brain regions yielded 141 differentially expressed cis-regulated probe sets and 206 trans-regulated probe sets. Comparing the present results from inbred alcohol-preferring vs. congenic P.NP rats to earlier results from the reciprocal congenic NP.P vs. inbred alcohol-nonpreferring rats demonstrated that 74 cis-regulated probe sets were differentially expressed in the same direction and with a consistent magnitude of difference in at least one brain region.
    [Show full text]
  • ECOP Shrna (H) Lentiviral Particles: Sc-89730-V
    SANTA CRUZ BIOTECHNOLOGY, INC. ECOP shRNA (h) Lentiviral Particles: sc-89730-V BACKGROUND APPLICATIONS ECOP (EGFR-coamplified and overexpressed protein), also known as VOPP1 ECOP shRNA (h) Lentiviral Particles is recommended for the inhibition of (vesicular, overexpressed in cancer, prosurvival protein 1) or GASP (glioblas- ECOP expression in human cells. toma-amplified secreted protein), is a 172 amino acid protein that is coam- plified with EGFR and overexpressed in multiple glioblastomas. Highly ex- SUPPORT REAGENTS pressed in ovary and thymus, ECOP is found at moderate levels in testis, Control shRNA Lentiviral Particles: sc-108080. Available as 200 µl frozen colon, small intestine, and spleen, and at low levels in liver, placenta and viral stock containing 1.0 x 106 infectious units of virus (IFU); contains an κ prostate. ECOP regulates NF B signaling and may have a role in resistance shRNA construct encoding a scrambled sequence that will not lead to the to apoptosis. The gene encoding ECOP maps to human chromosome 7, which specific degradation of any known cellular mRNA. houses over 1,000 genes, comprises nearly 5% of the human genome and has been linked to Osteogenesis imperfecta, Pendred syndrome, Lissencephaly, RT-PCR REAGENTS Citrullinemia and Shwachman-Diamond syndrome. Semi-quantitative RT-PCR may be performed to monitor ECOP gene expres- REFERENCES sion knockdown using RT-PCR Primer: ECOP (h)-PR: sc-89730-PR (20 µl). Annealing temperature for the primers should be 55-60° C and the extension 1. Tsipouras, P., Myers, J.C., Ramirez, F. and Prockop, D.J. 1983. Restriction temperature should be 68-72° C.
    [Show full text]
  • Identification and Analysis of Programmed Cell Death Genes in Drosophila Melanogaster and Human Cancer Using Biotnformatic Analysis of Gene Expression Data
    IDENTIFICATION AND ANALYSIS OF PROGRAMMED CELL DEATH GENES IN DROSOPHILA MELANOGASTER AND HUMAN CANCER USING BIOTNFORMATIC ANALYSIS OF GENE EXPRESSION DATA by ERBSf DAEL PLEASANCE B.Sc, The University of British Columbia, 2000 A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES (Medical Genetics) THE UNIVERSITY OF BRITISH COLUMBIA December 2005 © Erin Dael Pleasance, 2005 Abstract Programmed cell death (PCD), or cell suicide, encompasses multiple pathways including apoptosis and autophagy and is essential for development, cellular homeostasis, and prevention of cancer cell growth. I describe here the development and use of bioinformatic methods to identify and analyze genes involved in PCD, both in the model organism Drosophila melanogaster and in human cancer, by analysis of large-scale gene expression data. An approach was developed to correctly identify genes from serial analysis of gene expression (SAGE) data, distinguish the set of genes not accessible to the SAGE method, and determine the optimal set of enzymes for Drosophila, C. elegans, and human SAGE library construction. In Drosophila metamorphosis the salivary gland undergoes autophagic PCD, whereby cellular components are engulfed and degraded by cytoplasmic vacuoles, with additional hallmarks of apoptosis. This is an excellent model in which to study the genes involved in PCD. Transcriptional profiling of this tissue by expressed sequence tags (ESTs) and serial analysis of gene expression (SAGE) identified many genes differentially regulated prior to cell death, including genes known to be death regulators, genes in related pathways, genes of no known function, and potentially novel unannotated genes.
    [Show full text]
  • High-Resolution Genomic Copy Number Profiling of Glioblastoma Multiforme by Single Nucleotide Polymorphism DNA Microarray
    Published OnlineFirst May 12, 2009; DOI: 10.1158/1541-7786.MCR-08-0270 Published Online First on May 12, 2009 High-Resolution Genomic Copy Number Profiling of Glioblastoma Multiforme by Single Nucleotide Polymorphism DNA Microarray Dong Yin,1 Seishi Ogawa,3 Norihiko Kawamata,1 Patrizia Tunici,2 Gaetano Finocchiaro,4 Marica Eoli,4 Christian Ruckert,6 Thien Huynh,1 Gentao Liu,2 Motohiro Kato,3 Masashi Sanada,3 Anna Jauch,5 Martin Dugas,6 Keith L. Black,2 and H. Phillip Koeffler1 1Division of Hematology/Oncology and 2Maxine Dunitz Neurosurgical Institute, Cedars-Sinai Medical Center, University of California at Los Angeles School of Medicine, Los Angeles, California; 3Regeneration Medicine of Hematopoiesis, University of Tokyo, School of Medicine, Tokyo, Japan; 4National Neurological Institute “C Besta,” Milan, Italy; 5Institute of Human Genetics, University Hospital Heidelberg, Germany; and 6Department of Medical Informatics and Biomathematics, University of Munster, Munster, Germany Abstract growth factor receptor/platelet-derived growth factor receptor Glioblastoma multiforme (GBM) is an extremely malignant α. Deletion of chromosome 6q26-27 often occurred (16 of 55 brain tumor. To identify new genomic alterations in GBM, samples). The minimum common deleted region included genomic DNA of tumor tissue/explants from 55 individuals PARK2, PACRG, QKI,and PDE10A genes. Further reverse and 6 GBM cell lines were examined using single nucleotide transcription Q-PCR studies showed that PARK2 expression polymorphism DNA microarray (SNP-Chip). Further gene was decreased in another collection of GBMs at a expression analysis relied on an additional 56 GBM samples. frequency of 61% (34 of 56) of samples. The 1p36.23 region SNP-Chip results were validated using several techniques, was deleted in 35% (19 of 55) of samples.
    [Show full text]
  • Bcl2l10 Induces Metabolic Alterations in Ovarian Cancer Cells by Regulating the TCA Cycle Enzymes SDHD and IDH1
    ONCOLOGY REPORTS 45: 47, 2021 Bcl2l10 induces metabolic alterations in ovarian cancer cells by regulating the TCA cycle enzymes SDHD and IDH1 SU‑YEON LEE, JINIE KWON and KYUNG‑AH LEE Department of Biomedical Science, College of Life Science, CHA University, Seongnam, Gyeonggi 13488, Republic of Korea Received September 29, 2020; Accepted February 3, 2021 DOI: 10.3892/or.2021.7998 Abstract. Bcl2‑like‑10 (Bcl2l10) has both oncogenic and and the results indicated that Bcl2l10 may serve as a potential tumor suppressor functions depending on the type of cancer. therapeutic target in ovarian cancer. It has been previously demonstrated that the suppression of Bcl2l10 in ovarian cancer SKOV3 and A2780 cells causes Introduction cell cycle arrest and enhances cell proliferation, indicating that Bcl2l10 is a tumor suppressor gene in ovarian cancer Over the past years, cutting‑edge research and advanced cells. The aim of the present study was to identify possible screening, surgical and therapeutic technologies have contrib‑ downstream target genes and investigate the underlying uted to increasing the 5‑year relative survival rate for all types mechanisms of action of Bcl2l10 in ovarian cancer cells. of cancer from 68 to 86% from 2010 to 2016 in adolescents in RNA sequencing (RNA‑Seq) was performed to obtain a list of the United States (1). Despite these advances, the 5‑year overall differentially expressed genes (DEGs) in Bcl2l10‑suppressed survival rate for advanced ovarian cancer remains 29% after SKOV3 and A2780 cells. The RNA‑Seq data were validated diagnosis, as determined by statistics from 2008 to 2014 in by reverse transcription‑quantitative PCR (RT‑qPCR) and the USA (2).
    [Show full text]
  • Analysis of Gene Expression in Two Large Schizophrenia Cohorts Identifies Multiple Changes Associated with Nerve Terminal Function
    Analysis of gene expression in two large schizophrenia cohorts identifies multiple changes associated with nerve terminal function Peter R Maycox1, Fiona Kelly1, Adam Taylor1, Stewart Bates1, Juliet Reid1, Ramya Logendra1, Michael R. Barnes1, Chris Larminie1, Neil Jones1, Mark Lennon1, Ceri Davies1, Jim J Hagan1, Carol A Scorer1, Claire Angelinetta2, Tariq Akbar2, Steven Hirsch2, Ann M. Mortimer2,3, Thomas R. E. Barnes2 Jackie de Belleroche2* Author affiliation: 1Computational Biology, Genetic and Proteomic Sciences, Statistical Sciences, Psychiatry CEDD, GlaxoSmithKline, New Frontiers Science Park, GlaxoSmithKline, Harlow, Essex, CM19 5AW, UK 2Division of Neuroscience and Mental Health, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK 3Academic Unit of Psychiatry, University of Hull, Hull HU6 7RX, UK *Corresponding author: Professor Jackie de Belleroche, Neurogenetics Group, Division of Neuroscience and Mental Health, Imperial College London, Hammersmith Hospital Campus, Burlington Danes Building, Du Cane Road, London W12 0NN, UK Telephone: +44 207 5946649 Fax: +44 207 5946548 Email_ [email protected] Key words: schizophrenia, gene expression, microarrays, synaptic vesicle recycling, synaptic transmission Running title: Schizophrenia microarrays identify synaptic markers ABSTRACT Schizophrenia is a severe psychiatric disorder with a world wide prevalence of 1%. The pathophysiology of the illness is not understood, but is thought to have a strong genetic component with some environmental influences on aetiology. To gain further insight into disease mechanism, we used microarray technology to determine the expression of over 30,000 mRNA transcripts in post-mortem tissue from a brain region associated with the pathophysiology of the disease (Brodmann Area 10: anterior prefrontal cortex) in 28 schizophrenic and 23 control patients.
    [Show full text]