DOCSLIB.ORG

A Human-Mouse Conserved Sex Bias in Amygdala Gene Expression Related to Circadian Clock and Energy Metabolism Li-Chun Lin, David a Lewis and Etienne Sibille*

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A Human-Mouse Conserved Sex Bias in Amygdala Gene Expression Related to Circadian Clock and Energy Metabolism Li-Chun Lin, David a Lewis and Etienne Sibille* Lin et al. Molecular Brain 2011, 4:18 http://www.molecularbrain.com/content/4/1/18 SHORTREPORT Open Access A human-mouse conserved sex bias in amygdala gene expression related to circadian clock and energy metabolism Li-Chun Lin, David A Lewis and Etienne Sibille* Abstract Background: Major depression affects twice as many women as men, but the underlying molecular mechanisms responsible for the heightened female vulnerability are not known. The amygdala, composed of heterogeneous subnuclei, participates in multiple functional circuits regulating emotional responses to stress. We hypothesized that sex differences in molecular structure may contribute to differential mood regulation and disease vulnerability. Findings: Using gene arrays followed by quantitative PCR validation, we compared the transcriptome profiles between sexes in human and mouse amygdala. We now report sexually dimorphic features of transcriptomes in the basolateral nucleus of the amygdala, and these features are highly conserved across species. A functional analysis of differential gene expression showed that mitochondrial-related gene groups were identified as the top biological pathways associated with sexual dimorphism in both species. Conclusions: These results suggest that the basolateral amygdala is a sexually dimorphic structure, featuring a regulatory cascade of mitochondrial function and circadian rhythm, potentially linked through sirtuins and hormone nuclear receptors. Hence, baseline differences in amygdalar circadian regulation of cellular metabolism may contribute to sex-related differences in mood regulation and vulnerability to major depression. Introduction amygdala complex improves microarray sensitivity to Males and females differ in behavior and brain structure, detect differences in gene expression by reducing sample as well as in prevalence of neuropsychiatric disorders. heterogeneity, and could facilitate the subsequent identi- The greater prevalence of major depression and rapid fication of genetic sex differences. To characterize sex cycling bipolar disorder in women [1,2] highlights the differences in the intrinsic molecular properties of the importance of studying potential mechanisms for sex amygdala, we examined the presence of sexually differences. Sex chromosome- and hormone-linked dimorphic patterns of gene expression in the basolateral genes may directly affect sexually dimorphic neurobiol- nuclei of the amygdala using postmortem samples from ogy [3]. For example, sex hormones regulate the size of control human subjects and mice. the medial amygdala and bed nucleus of the stria termi- nalis during development [4]. The lateral (LA) and basal Methods (basolateral; throughout the article we refer to human Human Brain Samples BA and mouse BLA) nuclei of the amygdala have been Postmortem amygdala samples from 12 healthy male identified as critical sites for learned fear and emotion controls and 12 healthy female controls (age = 39-64 regulation in healthy subjects and in patients with mood years) were obtained from the University of Pittsburgh disorders [5,6], but little is known about their sexual Brain Donation Program (Table 1), as previously dimorphism. The isolation of subnuclei from the described [6]. Male and female groups were matched in pairs as closely as possible on the basis of RNA integrity, * Correspondence: [email protected] pH,post-morteminterval,age,andsex,andthegroups Translational Neuroscience Program, Department of Psychiatry, and Center did not differ on any of these parameters (p > 0.05; ’ for Neuroscience, 3811 O Hara Street, University of Pittsburgh, Pittsburgh, PA Table 1). All subjects died suddenly without prolonged 15213, USA © 2011 Lin 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. Lin et al. Molecular Brain 2011, 4:18 Page 2 of 6 http://www.molecularbrain.com/content/4/1/18 Table 1 Characteristics of matched human subjects Female Group Male Group Age Interval Age Interval Pair Case (Years) Race (Hours) pH RIN Case (Years) Race (Hours) pH RIN 1 HU1282 39 W 24.5 6.8 7.5 HU604 39 W 19.3 7.1 8.6 2 HU1092 40 B 16.6 6.8 8.0 HU1047 43 W 12.0 6.6 9.0 3 HU627a,b 43 B 14.1 7.1 7.0 HU857a,b 48 W 16.6 6.7 8.9 4 HU567 46 W 15.0 6.8 8.9 HU1067b 49 W 6.0 6.6 8.2 5 HU1280a 50 W 23.5 6.7 7.7 HU1086a,b 51 W 24.2 6.8 8.1 6 HU1391a,b 51 W 7.8 6.6 7.1 HU852a,b 54 W 8.0 6.8 9.1 7 HU686 52 W 22.6 7.1 8.5 HU1031b 53 W 23.2 6.8 8.9 8 HU575 55 B 11.3 6.8 9.6 HU1122 55 W 15.4 6.7 7.9 9 HU1247 58 W 22.7 6.4 8.4 HU685 56 W 14.0 6.6 8.0 10 HU1318 58 W 18.8 6.7 7.4 HU852 54 W 8.0 6.8 9.1 11 HU568 60 W 9.5 6.9 8.7 HU551 61 W 16.4 6.6 8.3 12 HU1466b 64 B 20.0 6.7 8.8 HU615 62 W 7.2 6.4 7.8 Matched samples processed for microarray: AVG 51.3 17.2 6.8 8.1 52.1 14.2 6.7 8.5 STDEV 8.1 5.7 0.2 0.8 6.7 6.2 0.2 0.5 Matched LA for final array analysis: AVG 52.4 17.9 6.8 8.4 52.4 13.5 6.7 8.4 STDEV 8.9 5.2 0.2 0.7 7.7 5.8 0.2 0.5 Matched BA for final array analysis: AVG 50.9 18.3 6.8 8.3 52.9 13.2 6.7 8.4 STDEV 7.8 5.5 0.2 0.7 8.7 4.4 0.2 0.5 Samples from LA (marked a) and BA (marked b) processed but excluded due to lower microarray quality control; W = Caucasian; B = African American; RIN = RNA integrity number; AVG = average; STDVE = standard deviation. agonal periods. See [6] for additional details. In contrast LMD 6500 laser microdissection system. All mice were to the light acetylcholinesterase (AChE)-stained LA maintained on a 12-hour light-dark cycle (lights on across species, human BA and mouse BLA contained 06:00 and lights off on 18:00) with ad libitum food and the heavy stained AChE-positive neuropil in the amyg- water. All procedures were approved by the University dala [7,8], implying that human BA and LA anatomically of Pittsburgh Institutional Animal Care and Use Com- correspond to the mouse BLA and LA according to the mittee (protocol # 0911014A-2, Animal Assurance # well-defined AChE boundaries. Hence, based on the dis- A3187-01). tribution of AChE activity and thionin staining, human lateral(LA)andbasal(BA)amygdalanucleiwere RNA extraction for Microarray Samples defined and separately harvested from 20-μmcryostat Total RNA was extracted using the RNeasy Plus Micro sections using a clean RNase-free pipette tip. All proce- Kit (QIAGEN, Valencia, CA) and processed for microar- dures were approved by the University of Pittsburgh’s ray analysis according to manufacturer’sprotocol.RNA Committee for the Oversight of Research Involving the samples were amplified with Illumina TotalPrep RNA Dead and Institutional Review Board for Biomedical Amplification Kit (for human tissue; Illumina, San Research. Diego, CA) and Nugen WT-Ovation Pico RNA Amplifi- cation System (for mouse tissue; NuGen, San Carlos, Mouse Brain Samples CA). The fragmented labeled cRNA samples were pro- Brains from adult C57BL/6J mice (n = 5 per sex; pooled cessed and hybridized to microarrays (Human HT-12v3 by 2 from 10 mice/group, 3 months of age) were har- and MouseWG-6v2.0 BeadChips). vested, fresh frozen and cryo-sectioned at 20 μmon UV-treated laser-capture microscopy slides (Leica Array Data Analysis Microsystems, Wetzlar, Germany). Sections on the slides For statistical analysis, the log2-transformed signal inten- were dried and stored at -80°C until used. Lateral (LA) sities of probe-sets that passed the initial expression to basolateral (BLA) amygdala nuclei were visualized filters (< 30% missing values, detection p-value > 0.1), with rapid thionin staining and dissected with the Leica were extracted and renormalized to the same mean and Lin et al. Molecular Brain 2011, 4:18 Page 3 of 6 http://www.molecularbrain.com/content/4/1/18 variance across arrays in order to control for any effects mouse LA (p< 6.72E-02). Assembly of RNA Polymerase of batch. Discovery threshold for male/female differ- III Complex was the only identified canonical pathway ences were applied in both species (two-tailed t-test p- in human LA and was at trend level (p < 0.1). value < 0.05; > 20% effect size). Functional analyses Within the mitochondrial-related functions, genes were performed on the identified gene list using the encoding enzymes of the mitochondrial electron trans- Ingenuity Pathways Knowledge Base [9] and compared port chain (ETC) consistently displayed high male-biased across species. expression (Figure 2A). Also, two genes coding for mito- chondria-localized sirtuins (SIRT3, SIRT5), which serve Real-time Quantitative PCR (qPCR) Data Analysis as metabolic regulators of circadian rhythms by utilizing Separate RNA samples were reverse-transcribed using NAD+ [10], also showed similar high male-biased expres- QScript cDNA super mix (QuantaBioscience, Gaithers- sion in human BA and mouse BLA. Of related interest burg, MD) using a combination of oligo (dT) and ran- was the identification of sex-biased circadian rhythm sig- dom primers. Each PCR reaction was performed in naling in human BA (IPA, p< 5.39E-04)andmouseLA triplicate and compared to three internal controls (p<3.54E-02), which included conserved high female- (GAPDH, beta-actin and cyclophilin).
Load more

Recommended publications
  • Supplement 1 Overview of Dystonia Genes
    Supplement 1 Overview of genes that may cause dystonia in children and adolescents Gene (OMIM) Disease name/phenotype Mode of inheritance 1: (Formerly called) Primary dystonias (DYTs): TOR1A (605204) DYT1: Early-onset generalized AD primary torsion dystonia (PTD) TUBB4A (602662) DYT4: Whispering dystonia AD GCH1 (600225) DYT5: GTP-cyclohydrolase 1 AD deficiency THAP1 (609520) DYT6: Adolescent onset torsion AD dystonia, mixed type PNKD/MR1 (609023) DYT8: Paroxysmal non- AD kinesigenic dyskinesia SLC2A1 (138140) DYT9/18: Paroxysmal choreoathetosis with episodic AD ataxia and spasticity/GLUT1 deficiency syndrome-1 PRRT2 (614386) DYT10: Paroxysmal kinesigenic AD dyskinesia SGCE (604149) DYT11: Myoclonus-dystonia AD ATP1A3 (182350) DYT12: Rapid-onset dystonia AD parkinsonism PRKRA (603424) DYT16: Young-onset dystonia AR parkinsonism ANO3 (610110) DYT24: Primary focal dystonia AD GNAL (139312) DYT25: Primary torsion dystonia AD 2: Inborn errors of metabolism: GCDH (608801) Glutaric aciduria type 1 AR PCCA (232000) Propionic aciduria AR PCCB (232050) Propionic aciduria AR MUT (609058) Methylmalonic aciduria AR MMAA (607481) Cobalamin A deficiency AR MMAB (607568) Cobalamin B deficiency AR MMACHC (609831) Cobalamin C deficiency AR C2orf25 (611935) Cobalamin D deficiency AR MTRR (602568) Cobalamin E deficiency AR LMBRD1 (612625) Cobalamin F deficiency AR MTR (156570) Cobalamin G deficiency AR CBS (613381) Homocysteinuria AR PCBD (126090) Hyperphelaninemia variant D AR TH (191290) Tyrosine hydroxylase deficiency AR SPR (182125) Sepiaterine reductase
    [Show full text]
  • Assessment of a Targeted Gene Panel for Identification of Genes Associated with Movement Disorders
    Supplementary Online Content Montaut S, Tranchant C, Drouot N, et al; French Parkinson’s and Movement Disorders Consortium. Assessment of a targeted gene panel for identification of genes associated with movement disorders. JAMA Neurol. Published online June 18, 2018. doi:10.1001/jamaneurol.2018.1478 eMethods. Supplemental methods. eTable 1. Name, phenotype and inheritance of the genes included in the panel. eTable 2. Probable pathogenic variants identified in a cohort of 23 patients with cerebellar ataxia using WES analysis. eTable 3. Negative cases in a cohort of 23 patients with cerebellar ataxia studied using WES analysis. eTable 4. Variants of unknown significance (VUSs) identified in the cohort. eFigure 1. Examples of pedigrees of cases with identified causative variants. eFigure 2. Pedigrees suggesting mendelian inheritance in negative cases. eFigure 3. Examples of pedigrees of cases with identified VUSs. eResults. Supplemental results. This supplementary material has been provided by the authors to give readers additional information about their work. © 2018 American Medical Association. All rights reserved. Downloaded From: https://jamanetwork.com/ on 09/26/2021 eMethods. Supplemental methods Patients selection In the multicentric, prospective study, patients were selected from 25 French, 1 Luxembourg and 1 Algerian tertiary MDs centers between September 2014 and July 2016. Inclusion criteria were patients (1) who had developed one or several chronic MDs (2) with an age of onset below 40 years and/or presence of a family history of MDs. Patients suffering from essential tremor, tic or Gilles de la Tourette syndrome, pure cerebellar ataxia or with clinical/paraclinical findings suggestive of an acquired cause were excluded.
    [Show full text]
  • 1 Metabolic Dysfunction Is Restricted to the Sciatic Nerve in Experimental
    Page 1 of 255 Diabetes Metabolic dysfunction is restricted to the sciatic nerve in experimental diabetic neuropathy Oliver J. Freeman1,2, Richard D. Unwin2,3, Andrew W. Dowsey2,3, Paul Begley2,3, Sumia Ali1, Katherine A. Hollywood2,3, Nitin Rustogi2,3, Rasmus S. Petersen1, Warwick B. Dunn2,3†, Garth J.S. Cooper2,3,4,5* & Natalie J. Gardiner1* 1 Faculty of Life Sciences, University of Manchester, UK 2 Centre for Advanced Discovery and Experimental Therapeutics (CADET), Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Sciences Centre, Manchester, UK 3 Centre for Endocrinology and Diabetes, Institute of Human Development, Faculty of Medical and Human Sciences, University of Manchester, UK 4 School of Biological Sciences, University of Auckland, New Zealand 5 Department of Pharmacology, Medical Sciences Division, University of Oxford, UK † Present address: School of Biosciences, University of Birmingham, UK *Joint corresponding authors: Natalie J. Gardiner and Garth J.S. Cooper Email: [email protected]; [email protected] Address: University of Manchester, AV Hill Building, Oxford Road, Manchester, M13 9PT, United Kingdom Telephone: +44 161 275 5768; +44 161 701 0240 Word count: 4,490 Number of tables: 1, Number of figures: 6 Running title: Metabolic dysfunction in diabetic neuropathy 1 Diabetes Publish Ahead of Print, published online October 15, 2015 Diabetes Page 2 of 255 Abstract High glucose levels in the peripheral nervous system (PNS) have been implicated in the pathogenesis of diabetic neuropathy (DN). However our understanding of the molecular mechanisms which cause the marked distal pathology is incomplete. Here we performed a comprehensive, system-wide analysis of the PNS of a rodent model of DN.
    [Show full text]
  • Supplemental Table 10
    Supplemental Table 10: Dietary Impact on the Heart Sulfhydrome DR/AL Accession Alternate Molecular Cysteine Spectral Protein Name Number ID Weight Residues Count Ratio P‐value Ig lambda‐2 chain C region P01844 Iglc2 11 kDa 3 C 16.000 0.00101 Gelsolin P13020 (+1) Gsn 86 kDa 7 C 11.130 0.00133 Glutamate‐‐cysteine ligase regulatory subunit O09172 Gclm 31 kDa 6 C 10.200 0.0307 Ig gamma‐3 chain C region P03987 44 kDa 10 C 7.636 0.0005 Ferritin heavy chain P09528 Fth1 21 kDa 3 C 6.182 0.02617 Antithrombin‐III P32261 Serpinc1 52 kDa 9 C 5.333 0.03116 Bisphosphoglycerate mutase P15327 Bpgm 30 kDa 3 C 4.645 0.01998 Vitamin D‐binding protein Q9QVP4 Gc 54 kDa 28 C 4.541 0.0206 Properdin P11680 Cfp 50 kDa 44 C 3.692 0.0227 Complement factor B P01867 (+1) Cfb 85 kDa 20 C 3.636 0.01126 Transforming growth factor beta‐1 P04202 Tgfb1 44 kDa 12 C 3.273 0.00601 Ferritin light chain 1 P29391 Ftl1 21 kDa 1 C 3.250 0.0204 Ig lambda‐1 chain C region Q9CPV4‐2 12 kDa 3 C 2.844 0.02618 Kininogen‐1 Q8K182 Kng1 73 kDa 19 C 2.840 0.01359 Beta‐2‐glycoprotein 1 Q01339 Apoh 39 kDa 23 C 2.691 0.00579 Complement C3 P01027 C3 186 kDa 27 C 2.556 0.00991 Complement factor I P02088 Cfi 67 kDa 40 C 2.324 0.02636 Ig heavy chain V region 102 P01750 13 kDa 3 C 16.200 0.1642 Afamin O89020 (+1) Afm 69 kDa 34 C 14.400 0.07963 Dehydrogenase/reductase SDR family member 11 Q3U0B3 Dhrs11 28 kDa 8 C 10.400 0.09207 Myosin light chain 4 P09541 Myl4 21 kDa 2 C 9.908 0.23919 Myeloperoxidase P11247 Mpo 81 kDa 16 C 8.800 0.40708 Myosin regulatory light chain 2, skeletal muscle isoform P97457
    [Show full text]
  • Association of Gene Ontology Categories with Decay Rate for Hepg2 Experiments These Tables Show Details for All Gene Ontology Categories
    Supplementary Table 1: Association of Gene Ontology Categories with Decay Rate for HepG2 Experiments These tables show details for all Gene Ontology categories. Inferences for manual classification scheme shown at the bottom. Those categories used in Figure 1A are highlighted in bold. Standard Deviations are shown in parentheses. P-values less than 1E-20 are indicated with a "0". Rate r (hour^-1) Half-life < 2hr. Decay % GO Number Category Name Probe Sets Group Non-Group Distribution p-value In-Group Non-Group Representation p-value GO:0006350 transcription 1523 0.221 (0.009) 0.127 (0.002) FASTER 0 13.1 (0.4) 4.5 (0.1) OVER 0 GO:0006351 transcription, DNA-dependent 1498 0.220 (0.009) 0.127 (0.002) FASTER 0 13.0 (0.4) 4.5 (0.1) OVER 0 GO:0006355 regulation of transcription, DNA-dependent 1163 0.230 (0.011) 0.128 (0.002) FASTER 5.00E-21 14.2 (0.5) 4.6 (0.1) OVER 0 GO:0006366 transcription from Pol II promoter 845 0.225 (0.012) 0.130 (0.002) FASTER 1.88E-14 13.0 (0.5) 4.8 (0.1) OVER 0 GO:0006139 nucleobase, nucleoside, nucleotide and nucleic acid metabolism3004 0.173 (0.006) 0.127 (0.002) FASTER 1.28E-12 8.4 (0.2) 4.5 (0.1) OVER 0 GO:0006357 regulation of transcription from Pol II promoter 487 0.231 (0.016) 0.132 (0.002) FASTER 6.05E-10 13.5 (0.6) 4.9 (0.1) OVER 0 GO:0008283 cell proliferation 625 0.189 (0.014) 0.132 (0.002) FASTER 1.95E-05 10.1 (0.6) 5.0 (0.1) OVER 1.50E-20 GO:0006513 monoubiquitination 36 0.305 (0.049) 0.134 (0.002) FASTER 2.69E-04 25.4 (4.4) 5.1 (0.1) OVER 2.04E-06 GO:0007050 cell cycle arrest 57 0.311 (0.054) 0.133 (0.002)
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • Growth and Gene Expression Profile Analyses of Endometrial Cancer Cells Expressing Exogenous PTEN
    [CANCER RESEARCH 61, 3741–3749, May 1, 2001] Growth and Gene Expression Profile Analyses of Endometrial Cancer Cells Expressing Exogenous PTEN Mieko Matsushima-Nishiu, Motoko Unoki, Kenji Ono, Tatsuhiko Tsunoda, Takeo Minaguchi, Hiroyuki Kuramoto, Masato Nishida, Toyomi Satoh, Toshihiro Tanaka, and Yusuke Nakamura1 Laboratories of Molecular Medicine [M. M-N., M. U., K. O., T. M., T. Ta., Y. N.] and Genome Database [T. Ts.], Human Genome Center, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan; Department of Obstetrics and Gynecology, School of Medicine, Kitasato University, Sagamihara 228-8555, Japan [H. K.]; Department of Obstetrics and Gynecology, Institute of Clinical Medicine, University of Tsukuba, Tsukuba 305-8576, Japan [M. N.]; and Department of Obstetrics and Gynecology, Ibaraki Seinan Central Hospital, Tsukuba 306-0433, Japan [T. S.] ABSTRACT Akt/protein kinase B, cell survival, and cell proliferation (8). Over- expression of PTEN can decrease cell proliferation and tumorigenicity The PTEN tumor suppressor gene encodes a multifunctional phospha- (9, 10), an observation attributed to the ability of PTEN to induce cell tase that plays an important role in inhibiting the phosphatidylinositol-3- cycle arrest and apoptosis (11, 12). kinase pathway and downstream functions that include activation of Akt/protein kinase B, cell survival, and cell proliferation. Enforced ex- Thus, lack of PTEN expression may affect a complex set of pression of PTEN in various cancer cell lines decreases cell proliferation transcriptional targets. However, no systematic assessment of PTEN- through arrest of the cell cycle, accompanied in some cases by induction regulated targets in cancer cells has been reported to date.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown Et Al
    US 20030082511A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown et al. (43) Pub. Date: May 1, 2003 (54) IDENTIFICATION OF MODULATORY Publication Classification MOLECULES USING INDUCIBLE PROMOTERS (51) Int. Cl." ............................... C12O 1/00; C12O 1/68 (52) U.S. Cl. ..................................................... 435/4; 435/6 (76) Inventors: Steven J. Brown, San Diego, CA (US); Damien J. Dunnington, San Diego, CA (US); Imran Clark, San Diego, CA (57) ABSTRACT (US) Correspondence Address: Methods for identifying an ion channel modulator, a target David B. Waller & Associates membrane receptor modulator molecule, and other modula 5677 Oberlin Drive tory molecules are disclosed, as well as cells and vectors for Suit 214 use in those methods. A polynucleotide encoding target is San Diego, CA 92121 (US) provided in a cell under control of an inducible promoter, and candidate modulatory molecules are contacted with the (21) Appl. No.: 09/965,201 cell after induction of the promoter to ascertain whether a change in a measurable physiological parameter occurs as a (22) Filed: Sep. 25, 2001 result of the candidate modulatory molecule. Patent Application Publication May 1, 2003 Sheet 1 of 8 US 2003/0082511 A1 KCNC1 cDNA F.G. 1 Patent Application Publication May 1, 2003 Sheet 2 of 8 US 2003/0082511 A1 49 - -9 G C EH H EH N t R M h so as se W M M MP N FIG.2 Patent Application Publication May 1, 2003 Sheet 3 of 8 US 2003/0082511 A1 FG. 3 Patent Application Publication May 1, 2003 Sheet 4 of 8 US 2003/0082511 A1 KCNC1 ITREXCHO KC 150 mM KC 2000000 so 100 mM induced Uninduced Steady state O 100 200 300 400 500 600 700 Time (seconds) FIG.
    [Show full text]
  • Gene Expression Profiling Analysis of Lung Adenocarcinoma
    Brazilian Journal of Medical and Biological Research (2016) 49(3): e4861, http://dx.doi.org/10.1590/1414-431X20154861 ISSN 1414-431X 1/11 Gene expression profiling analysis of lung adenocarcinoma H. Xu1,2,J.Ma1,J.Wu1, L. Chen1,F.Sun1,C.Qu1, D. Zheng1 and S. Xu1 1Department of Thoracic Surgery, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang, China 2Laboratory of Medical Genetics, Harbin Medical University, Harbin, Heilongjiang, China Abstract The present study screened potential genes related to lung adenocarcinoma, with the aim of further understanding disease pathogenesis. The GSE2514 dataset including 20 lung adenocarcinoma and 19 adjacent normal tissue samples from 10 patients with lung adenocarcinoma aged 45-73 years was downloaded from Gene Expression Omnibus. Differentially expressed genes (DEGs) between the two groups were screened using the t-test. Potential gene functions were predicted using functional and pathway enrichment analysis, and protein-protein interaction (PPI) networks obtained from the STRING database were constructed with Cytoscape. Module analysis of PPI networks was performed through MCODE in Cytoscape. In total, 535 upregulated and 465 downregulated DEGs were identified. These included ATP5D, UQCRC2, UQCR11 and genes encoding nicotinamide adenine dinucleotide (NADH), which are mainly associated with mitochondrial ATP synthesis coupled electron transport, and which were enriched in the oxidative phosphorylation pathway. Other DEGs were associated with DNA replication (PRIM1, MCM3, and RNASEH2A), cell surface receptor-linked signal transduction and the enzyme-linked receptor protein signaling pathway (MAPK1, STAT3, RAF1, and JAK1), and regulation of the cytoskeleton and phosphatidylinositol signaling system (PIP5K1B, PIP5K1C, and PIP4K2B). Our findings suggest that DEGs encoding subunits of NADH, PRIM1, MCM3, MAPK1, STAT3, RAF1, and JAK1 might be associated with the development of lung adenocarcinoma.
    [Show full text]
  • Laser Microdissection-Based Microproteomics of the Hippocampus of a Rat Epilepsy Model Reveals Regional Diferences in Protein Abundances Amanda M
    www.nature.com/scientificreports OPEN Laser microdissection-based microproteomics of the hippocampus of a rat epilepsy model reveals regional diferences in protein abundances Amanda M. do Canto 1,9, André S. Vieira 2,9, Alexandre H.B. Matos1,9, Benilton S. Carvalho3,9, Barbara Henning 1,9, Braxton A. Norwood4,5, Sebastian Bauer5,6, Felix Rosenow5,6, Rovilson Gilioli7, Fernando Cendes 8,9 & Iscia Lopes-Cendes 1,9* Mesial temporal lobe epilepsy (MTLE) is a chronic neurological disorder afecting almost 40% of adult patients with epilepsy. Hippocampal sclerosis (HS) is a common histopathological abnormality found in patients with MTLE. HS is characterised by extensive neuronal loss in diferent hippocampus sub- regions. In this study, we used laser microdissection-based microproteomics to determine the protein abundances in diferent regions and layers of the hippocampus dentate gyrus (DG) in an electric stimulation rodent model which displays classical HS damage similar to that found in patients with MTLE. Our results indicate that there are diferences in the proteomic profles of diferent layers (granule cell and molecular), as well as diferent regions, of the DG (ventral and dorsal). We have identifed new signalling pathways and proteins present in specifc layers and regions of the DG, such as PARK7, RACK1, and connexin 31/gap junction. We also found two major signalling pathways that are common to all layers and regions: infammation and energy metabolism. Finally, our results highlight the utility of high-throughput microproteomics and spatial-limited isolation of tissues in the study of complex disorders to fully appreciate the large biological heterogeneity present in diferent cell populations within the central nervous system.
    [Show full text]
  • The Diabetic Brain and Cognition
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by SZTE Publicatio Repozitórium - SZTE - Repository of Publications J Neural Transm DOI 10.1007/s00702-017-1763-2 NEUROLOGY AND PRECLINICAL NEUROLOGICAL STUDIES - REVIEW ARTICLE The diabetic brain and cognition 1 16 2 3 Peter Riederer • Amos D. Korczyn • Sameh S. Ali • Ovidiu Bajenaru • 4 5 7 Mun Seong Choi • Michael Chopp • Vesna Dermanovic-Dobrota • 8,9,10 11 13,14,15 Edna Gru¨nblatt • Kurt A. Jellinger • Mohammad Amjad Kamal • 12 17 19 Warda Kamal • Jerzy Leszek • Tanja Maria Sheldrick-Michel • 20 18 6 21 Gohar Mushtaq • Bernard Meglic • Rachel Natovich • Zvezdan Pirtosek • 22 23 24 Martin Rakusa • Melita Salkovic-Petrisic • Reinhold Schmidt • 25 26 27 Angelika Schmitt • G. Ramachandra Sridhar • La´szlo´ Ve´csei • 28 29 5,29 6 Zyta Beata Wojszel • Hakan Yaman • Zheng G. Zhang • Tali Cukierman-Yaffe Received: 1 June 2017 / Accepted: 13 July 2017 Ó Springer-Verlag GmbH Austria 2017 Abstract The prevalence of both Alzheimer’s disease (AD) Congress on Vascular Disorders and on literature search and vascular dementia (VaD) is increasing with the aging of using PUBMED, it can be concluded that T2DM is a risk the population. Studies from the last several years have factor for both, AD and VaD, based on a pathology of glu- shown that people with diabetes have an increased risk for cose utilization. This pathology is the consequence of a dementia and cognitive impairment. Therefore, the authors disturbance of insulin-related mechanisms leading to brain of this consensus review tried to elaborate on the role of insulin resistance.
    [Show full text]
  • Coupling Mitochondrial Respiratory Chain to Cell Death: an Essential Role of Mitochondrial Complex I in the Interferon-B and Retinoic Acid-Induced Cancer Cell Death
    Cell Death and Differentiation (2007) 14, 327–337 & 2007 Nature Publishing Group All rights reserved 1350-9047/07 $30.00 www.nature.com/cdd Coupling mitochondrial respiratory chain to cell death: an essential role of mitochondrial complex I in the interferon-b and retinoic acid-induced cancer cell death G Huang1, Y Chen1,HLu1 and X Cao*,1 Combination of retinoic acids (RAs) and interferons (IFNs) has synergistic apoptotic effects and is used in cancer treatment. However, the underlying mechanisms remain unknown. Here, we demonstrate that mitochondrial respiratory chain (MRC) plays an essential role in the IFN-b/RA-induced cancer cell death. We found that IFN-b/RA upregulates the expression of MRC complex subunits. Mitochondrial–nuclear translocation of these subunits was not observed, but overproduction of reactive oxygen species (ROS), which causes loss of mitochondrial function, was detected upon IFN-b/RA treatment. Knockdown of GRIM-19 (gene associated with retinoid-interferon-induced mortality-19) and NDUFS3 (NADH dehydrogenase (ubiquinone) Fe-S protein 3), two subunits of MRC complex I, by siRNA in two cancer cell lines conferred resistance to IFN-b/RA-induced apoptosis and reduced ROS production. In parallel, expression of late genes induced by IFN-b/RA that are directly involved in growth inhibition and cell death was also repressed in the knockdown cells. Our data suggest that the MRC regulates IFN-b/RA-induced cell death by modulating ROS production and late gene expression. Cell Death and Differentiation (2007) 14, 327–337.
    [Show full text]