DOCSLIB.ORG

PER1 Is a Prognostic Biomarker and Correlated with Immune Infiltrates

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
PER1 Is a Prognostic Biomarker and Correlated with Immune Infiltrates ORIGINAL RESEARCH published: 17 June 2021 doi: 10.3389/fgene.2021.697471 PER1 Is a Prognostic Biomarker and Correlated With Immune Infiltrates in Ovarian Cancer Mali Chen 1†, Lili Zhang 1†, Xiaolong Liu 2, Zhen Ma 2 and Ling Lv 1* 1 Department of Obstetrics, Gansu Province Maternity and Child-Care Hospital, Lanzhou, China, 2 Department of Surgical Oncology, Lanzhou University Second Hospital, Lanzhou, China Background: Period circadian protein homolog 1 (PER1) is an important component of the biorhythm molecular oscillation system and plays an important part in the development and progression of mammalian cancer. However, the correlations of PER1 with prognosis and tumor-infiltrating lymphocytes in ovarian cancer (OV) remain unclear. Methods: The Oncomine and TIMER databases were used to examine the expression Edited by: of PER1 in OV. Kaplan–Meier Plotter and PrognoScan were used to evaluate the Longxiang Xie, Henan University, China relationship between PER1 and prognosis. Kaplan–Meier Plotter was used to analyze Reviewed by: the relationships between PER1 and clinicopathological features of OV patients. The Gan Luan, relationship between PER1 expression and immune infiltration in OV was investigated New Jersey Institute of Technology, United States using the TIMER database and CIBERSORT algorithm. The STRING database was used Peijie Wu, to analyze PER1-related protein functional groups, the GeneMANIA online tool was used Chengdu University of Traditional to analyze gene groups with similar functions to those of PER1, and Network Analyst was Chinese Medicine, China used to identify transcription factors that regulate PER1. The correlation between PER1 *Correspondence: Ling Lv and immunoinvasion of OV was analyzed using TIMER. Finally, quantitative real-time [email protected] polymerase chain reaction (qRT-PCR) was performed to detect PER1 expression. †These authors have contributed Results: PER1 was differentially expressed in different cancer tissues, and its expression equally to this work and share first authorship in various OV subtypes was lower than that in normal ovarian tissue. OV patients with low PER1 expression had a reduced overall survival rate. Decreased PER1 expression in Specialty section: stage 1 and stage 1+2 OV patients was related to poor prognosis, while increased PER1 This article was submitted to + Cancer Genetics and Oncogenomics, expression in stage 3 4 patients and TP53 mutation were related to poor overall survival a section of the journal and progression-free survival. We identified eight genes whose expression was strongly Frontiers in Genetics correlated with that of PER1, as well as four transcription factors that regulate PER1. Received: 19 April 2021 In OV, PER1 expression levels were positively correlated with infiltration levels of cells Accepted: 19 May 2021 Published: 17 June 2021 including neutrophils, regulatory T cells, and M2 macrophages, and closely related to a Citation: variety of immune markers. Reduced expression of PER1 was significantly associated Chen M, Zhang L, Liu X, Ma Z and with poor overall survival. Lv L (2021) PER1 Is a Prognostic Biomarker and Correlated With Conclusion: These findings suggest that PER1 could be used as a prognostic Immune Infiltrates in Ovarian Cancer. biomarker to determine prognosis and immune infiltration in OV patients. Front. Genet. 12:697471. doi: 10.3389/fgene.2021.697471 Keywords: ovarian cancer, PER1, bioinformatics, diagnosis and prognosis biomarkers, immune infiltrates Frontiers in Genetics | www.frontiersin.org 1 June 2021 | Volume 12 | Article 697471 Chen et al. PER1 in OV INTRODUCTION MATERIALS AND METHODS Ovarian cancer (OV) is the fifth leading cause of cancer-related Tissue Samples deaths in women worldwide (Dong et al., 2014). Despite advances We collected 60 OV cancerous and paracancerous tissue in diagnosis, surgery, chemotherapy, and radiation therapy, OV samples together with associated clinical information from continues to have a poor survival rate and the highest mortality Gansu Provincial Maternal and Child Health Hospital. None of rate among gynecological cancers. OV patients diagnosed at an the subjects underwent radiotherapy or chemotherapy prior to advanced stage have poor prognosis and high recurrence rates. the surgery. All patient materials were obtained with informed Determining effective markers for early diagnosis and prognosis consent, and this study was carried out with the approval of the is thus essential to reduce mortality and ensure effective therapy Clinical Research Ethics Committees. for OV (Bowtell et al., 2015). Period circadian protein homolog 1 (PER1) is a core gene qRT-PCR involved in circadian rhythm. It is located on the short arm TRIzol reagent (Invitrogen, Carlsbad, CA, USA) was used for of human chromosome 17 (17p12–p13.1) and has a total total RNA extraction from tissues. RNA was reverse transcribed ∼ length of 16kb (Badiu, 2003). PER1 also has an important into cDNA using a RevertAid First Strand cDNA Synthesis Kit effect on the occurrence and development of tumors. Its (Thermo-Fisher Scientific, Waltham, MA, USA). Subsequently, main functions involve regulating the body’s circadian rhythm, with the cDNA as the template, qRT-PCR was performed regulating the cell cycle, and promoting DNA damage repair. using SYBR Premix Ex TaqTM (TaKaRa, Otsu, Shiga, Japan). Many studies have shown that PER1 directly regulates levels Relative gene expression was calculated by the 2–Ct method. of reactive oxygen species in multiple organs and regulates key GAPDH served as the internal reference. The primer sequences effectors of energy substrate utilization in response to daily were as follows (“F” represents “forward”; “R” represents behavioral rhythms and oxidative stress, thereby maintaining “reverse”). GAPDH: 5′-AGAAGGCTGGGGCTCATTTG- the daily rhythms of mitochondrial morphology and function 3′(F), 5′AGGGGCCATCCACAGTCTTC-3′(R); PER1: ′ ′ (Sun et al., 2020). Changes in expression levels of PER1 5 -CTGCTACAGGCACGTTCAAG-3 (F), 5′-CTCAGGG have been found in some cancers, including pancreatic cancer ACCAAGGCTAGTG-3′(R). (Guo et al., 2020), oral squamous cell carcinoma (Yang et al., 2020), endometrial cancer (Wang et al., 2020), colorectal cancer (Holipah and Kuroda, 2020), and non-small-cell carcinoma Oncomine Database Analysis (Lin et al., 2020), but this has not been studied in OV. It Oncomine is currently the world’s largest oncogene chip database has also been reported that the occurrence and development and data mining platform, containing 715 gene expression of cancer, as well as its prognosis and treatment outcomes, datasets and 86,733 tumor and normal tissue samples. Here, the are closely related to the abnormal expression of certain Oncomine database was used to detect the differential expression circadian clock genes (Cao et al., 2009; Sahar and Sassone- of PER1 between OV tissues and normal ovarian tissues. The data Corsi, 2009; Yang et al., 2009). Based on these findings, we type was mRNA, and differential genes were identified by t-test propose that PER1 could be used as a prognostic indicator with the following screening criteria: fold change >2, P < 0.05. in OV. We also note that circadian rhythm is closely related to GEPIA Database Analysis the human immune system (Lange et al., 2010). Abnormalities GEPIA is a newly developed bioinformatics platform for analysis of the biological clock contribute to exhaustion of T cells and processing of transcriptome data from The Cancer Genome and overall upregulation of immunosuppressive molecules Atlas (TCGA) and GTEx databases. We analyzed the correlations (including programmed death ligand 1 [PD-L1] and cytotoxic T between PER1 and other genes using the “Correlation Analysis” lymphocyte-associated antigen 4 [CTLA-4]); however, the impact module of GEPIA with P < 0.05 as the screening criterion. of PER1 on the immune environment of OV remains unknown. In this research, we analyzed the expression levels of PER1 in OV, using the online tools TIMER and Oncomine, TIMER Database and studied the relationships between PER1 expression and TIMER (https://cistrome.shinyapps.io/timer/) is a database for clinicopathological parameters. The prognostic value of comprehensive analysis of tumor immunity. We used the PER1 expression in OV was determined using Kaplan–Meier “Gene” module to analyze the correlations between PER1 gene Plotter and the PrognoScan database. Quantitative real-time expression and infiltration of immune cells, including CD4+ PCR (qRT-PCR) was used to detect PER1 expression, and T cells, CD8+ T cells, B cells, neutrophils, macrophages, and patients were divided into high and low PER1 expression dendritic cells. Immune cells included in the new version groups for survival analysis. In addition, we used the of TIMER showed increased infiltration levels, as presented TIMER database to determine the correlations between in the results section. We used the “Diff Exp” module to PER1 and immune cell infiltration. Our findings reveal detect the differential expression of the PER1 gene in different an important role of PER1 in OV and also clarify the tumor tissues and normal tissues. Then, we used TIMER to potential relationship of PER1 with tumor immunity and detect the correlations between PER1 and immune markers of its underlying mechanism. immune cells. Frontiers in Genetics | www.frontiersin.org 2 June 2021 | Volume 12 | Article 697471 Chen et al. PER1 in OV Survival Analysis and Prognosis Evaluation control samples were available for OV (Figure 1B). We further We used Kaplan–Meier
Load more

Recommended publications
  • Gene Symbol Gene Description ACVR1B Activin a Receptor, Type IB
    Table S1. Kinase clones included in human kinase cDNA library for yeast two-hybrid screening Gene Symbol Gene Description ACVR1B activin A receptor, type IB ADCK2 aarF domain containing kinase 2 ADCK4 aarF domain containing kinase 4 AGK multiple substrate lipid kinase;MULK AK1 adenylate kinase 1 AK3 adenylate kinase 3 like 1 AK3L1 adenylate kinase 3 ALDH18A1 aldehyde dehydrogenase 18 family, member A1;ALDH18A1 ALK anaplastic lymphoma kinase (Ki-1) ALPK1 alpha-kinase 1 ALPK2 alpha-kinase 2 AMHR2 anti-Mullerian hormone receptor, type II ARAF v-raf murine sarcoma 3611 viral oncogene homolog 1 ARSG arylsulfatase G;ARSG AURKB aurora kinase B AURKC aurora kinase C BCKDK branched chain alpha-ketoacid dehydrogenase kinase BMPR1A bone morphogenetic protein receptor, type IA BMPR2 bone morphogenetic protein receptor, type II (serine/threonine kinase) BRAF v-raf murine sarcoma viral oncogene homolog B1 BRD3 bromodomain containing 3 BRD4 bromodomain containing 4 BTK Bruton agammaglobulinemia tyrosine kinase BUB1 BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1B BUB1 budding uninhibited by benzimidazoles 1 homolog beta (yeast) C9orf98 chromosome 9 open reading frame 98;C9orf98 CABC1 chaperone, ABC1 activity of bc1 complex like (S. pombe) CALM1 calmodulin 1 (phosphorylase kinase, delta) CALM2 calmodulin 2 (phosphorylase kinase, delta) CALM3 calmodulin 3 (phosphorylase kinase, delta) CAMK1 calcium/calmodulin-dependent protein kinase I CAMK2A calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CAMK2B calcium/calmodulin-dependent
    [Show full text]
  • SHARP1 (BHLHE41) Mouse Monoclonal Antibody [Clone ID: OTI3H4] Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for TA806354 SHARP1 (BHLHE41) Mouse Monoclonal Antibody [Clone ID: OTI3H4] Product data: Product Type: Primary Antibodies Clone Name: OTI3H4 Applications: IHC, WB Recommended Dilution: WB 1:2000, IHC 1:150 Reactivity: Human Host: Mouse Isotype: IgG1 Clonality: Monoclonal Immunogen: Human recombinant protein fragment corresponding to amino acids 1-297 of human BHLHE41(NP_110389) produced in E.coli. Formulation: PBS (PH 7.3) containing 1% BSA, 50% glycerol and 0.02% sodium azide. Concentration: 1 mg/ml Purification: Purified from mouse ascites fluids or tissue culture supernatant by affinity chromatography (protein A/G) Conjugation: Unconjugated Storage: Store at -20°C as received. Stability: Stable for 12 months from date of receipt. Predicted Protein Size: 50.3 kDa Gene Name: basic helix-loop-helix family member e41 Database Link: NP_110389 Entrez Gene 79365 Human Q9C0J9 Background: This gene encodes a basic helix-loop-helix protein expressed in various tissues. The encoded protein can interact with ARNTL or compete for E-box binding sites in the promoter of PER1 and repress CLOCK/ARNTL's transactivation of PER1. This gene is believed to be involved in the control of circadian rhythm and cell differentiation. Defects in this gene are associated with the short sleep phenotype. [provided by RefSeq, Feb 2014] This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 SHARP1 (BHLHE41) Mouse Monoclonal Antibody [Clone ID: OTI3H4] – TA806354 Synonyms: BHLHB3; DEC2; hDEC2; SHARP1 Protein Families: Transcription Factors Protein Pathways: Circadian rhythm - mammal Product images: HEK293T cells were transfected with the pCMV6- ENTRY control (Left lane) or pCMV6-ENTRY BHLHE41 ([RC206882], Right lane) cDNA for 48 hrs and lysed.
    [Show full text]
  • Paul Hardin, Ph.D. John W
    Department of Biology The College of Arts + Sciences | Indiana University Bloomington About Paul Hardin Distinguished Alumni Award Lecture Thu., Oct. 18, 2018 • 4 to 5 pm • Myers Hall 130 Paul Hardin, Ph.D. John W. Lyons Jr. ’59 Chair in Biology, Texas A&M University Genetic architecture underlying circadian clock initiation, maintenance, and output in Drosophila Circadian clocks drive daily rhythms in metabolism, physiology, and behavior in organisms ranging from cyanobacteria to humans. The identification and analysis of “clock genes” in Drosophila revealed that circadian timekeeping is based on a transcriptional feedback loop Paul Hardin studied the development of the sea in which CLOCK-CYCLE (CLK-CYC) heterodimers activate transcription of their feedback urchin embryo in William Klein’s lab at Indiana repressors PERIOD (PER) and TIMELESS (TIM). Subsequent studies revealed that similar University, from where he received his Ph.D. in transcriptional feedback loops keep circadian time in all eukaryotes and, in the case of 1987. He did his postdoctoral fellowship with animals, that these feedback loops are comprised of conserved components. The “core” Michael Rosbash at Brandeis University, working feedback loop described above operates in conjunction with an “interlocked” feedback on the circadian rhythms of the fruit fly, Drosophila loop in animals to drive rhythmic transcription of hundreds of genes that are maximally melanogaster. His work with Michael Rosbash and expressed at different phases of the circadian cycle. These feedback loops operate in many, Jeff Hall has been instrumental to our understanding but not all, tissues in flies including the brain pacemaker neurons that control rest:activity of how circadian rhythms affect a myriad of rhythms.
    [Show full text]
  • Supervised Group Lasso with Applications to Microarray Data Analysis
    SUPERVISED GROUP LASSO WITH APPLICATIONS TO MICROARRAY DATA ANALYSIS Shuangge Ma1, Xiao Song2, and Jian Huang3 1Department of Epidemiology and Public Health, Yale University 2Department of Health Administration, Biostatistics and Epidemiology, University of Georgia 3Departments of Statistics and Actuarial Science, and Biostatistics, University of Iowa March 2007 The University of Iowa Department of Statistics and Actuarial Science Technical Report No. 375 1 Supervised group Lasso with applications to microarray data analysis Shuangge Ma¤1 Xiao Song 2and Jian Huang 3 1 Department of Epidemiology and Public Health, Yale University, New Haven, CT 06520, USA 2 Department of Health Administration, Biostatistics and Epidemiology, University of Georgia, Athens, GA 30602, USA 3 Department of Statistics and Actuarial Science, University of Iowa, Iowa City, IA 52242, USA Email: Shuangge Ma¤- [email protected]; Xiao Song - [email protected]; Jian Huang - [email protected]; ¤Corresponding author Abstract Background: A tremendous amount of e®orts have been devoted to identifying genes for diagnosis and prognosis of diseases using microarray gene expression data. It has been demonstrated that gene expression data have cluster structure, where the clusters consist of co-regulated genes which tend to have coordinated functions. However, most available statistical methods for gene selection do not take into consideration the cluster structure. Results: We propose a supervised group Lasso approach that takes into account the cluster structure in gene expression data for gene selection and predictive model building. For gene expression data without biological cluster information, we ¯rst divide genes into clusters using the K-means approach and determine the optimal number of clusters using the Gap method.
    [Show full text]
  • Watsonjn2018.Pdf (1.780Mb)
    UNIVERSITY OF CENTRAL OKLAHOMA Edmond, Oklahoma Department of Biology Investigating Differential Gene Expression in vivo of Cardiac Birth Defects in an Avian Model of Maternal Phenylketonuria A THESIS SUBMITTED TO THE GRADUATE FACULTY In partial fulfillment of the requirements For the degree of MASTER OF SCIENCE IN BIOLOGY By Jamie N. Watson Edmond, OK June 5, 2018 J. Watson/Dr. Nikki Seagraves ii J. Watson/Dr. Nikki Seagraves Acknowledgements It is difficult to articulate the amount of gratitude I have for the support and encouragement I have received throughout my master’s thesis. Many people have added value and support to my life during this time. I am thankful for the education, experience, and friendships I have gained at the University of Central Oklahoma. First, I would like to thank Dr. Nikki Seagraves for her mentorship and friendship. I lucked out when I met her. I have enjoyed working on this project and I am very thankful for her support. I would like thank Thomas Crane for his support and patience throughout my master’s degree. I would like to thank Dr. Shannon Conley for her continued mentorship and support. I would like to thank Liz Bullen and Dr. Eric Howard for their training and help on this project. I would like to thank Kristy Meyer for her friendship and help throughout graduate school. I would like to thank my committee members Dr. Robert Brennan and Dr. Lilian Chooback for their advisement on this project. Also, I would like to thank the biology faculty and staff. I would like to thank the Seagraves lab members: Jailene Canales, Kayley Pate, Mckayla Muse, Grace Thetford, Kody Harvey, Jordan Guffey, and Kayle Patatanian for their hard work and support.
    [Show full text]
  • Casein Kinase 1 Isoforms in Degenerative Disorders
    CASEIN KINASE 1 ISOFORMS IN DEGENERATIVE DISORDERS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Theresa Joseph Kannanayakal, M.Sc., M.S. * * * * * The Ohio State University 2004 Dissertation Committee: Approved by Professor Jeff A. Kuret, Adviser Professor John D. Oberdick Professor Dale D. Vandre Adviser Professor Mike X. Zhu Biophysics Graduate Program ABSTRACT Casein Kinase 1 (CK1) enzyme is one of the largest family of Serine/Threonine protein kinases. CK1 has a wide distribution spanning many eukaryotic families. In cells, its kinase activity has been found in various sub-cellular compartments enabling it to phosphorylate many proteins involved in cellular maintenance and disease pathogenesis. Tau is one such substrate whose hyperphosphorylation results in degeneration of neurons in Alzheimer’s disease (AD). AD is a slow neuroprogessive disorder histopathologically characterized by Granulovacuolar degeneration bodies (GVBs) and intraneuronal accumulation of tau in Neurofibrillary Tangles (NFTs). The level of CK1 isoforms, CK1α, CK1δ and CK1ε has been shown to be elevated in AD. Previous studies of the correlation of CK1δ with lesions had demonstrated its importance in tau hyperphosphorylation. Hence we investigated distribution of CK1α and CK1ε with the lesions to understand if they would play role in tau hyperphosphorylation similar to CK1δ. The kinase results were also compared with lesion correlation studies of peptidyl cis/trans prolyl isomerase (Pin1) and caspase-3. Our results showed that among the enzymes investigated, CK1 isoforms have the greatest extent of colocalization with the lesions. We have also investigated the distribution of CK1α with different stages of NFTs that follow AD progression.
    [Show full text]
  • Transcriptional Control of Tissue-Resident Memory T Cell Generation
    Transcriptional control of tissue-resident memory T cell generation Filip Cvetkovski Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2019 © 2019 Filip Cvetkovski All rights reserved ABSTRACT Transcriptional control of tissue-resident memory T cell generation Filip Cvetkovski Tissue-resident memory T cells (TRM) are a non-circulating subset of memory that are maintained at sites of pathogen entry and mediate optimal protection against reinfection. Lung TRM can be generated in response to respiratory infection or vaccination, however, the molecular pathways involved in CD4+TRM establishment have not been defined. Here, we performed transcriptional profiling of influenza-specific lung CD4+TRM following influenza infection to identify pathways implicated in CD4+TRM generation and homeostasis. Lung CD4+TRM displayed a unique transcriptional profile distinct from spleen memory, including up-regulation of a gene network induced by the transcription factor IRF4, a known regulator of effector T cell differentiation. In addition, the gene expression profile of lung CD4+TRM was enriched in gene sets previously described in tissue-resident regulatory T cells. Up-regulation of immunomodulatory molecules such as CTLA-4, PD-1, and ICOS, suggested a potential regulatory role for CD4+TRM in tissues. Using loss-of-function genetic experiments in mice, we demonstrate that IRF4 is required for the generation of lung-localized pathogen-specific effector CD4+T cells during acute influenza infection. Influenza-specific IRF4−/− T cells failed to fully express CD44, and maintained high levels of CD62L compared to wild type, suggesting a defect in complete differentiation into lung-tropic effector T cells.
    [Show full text]
  • Loss of Tcf7 Diminishes Hematopoietic Stem&Sol
    Letters to the Editor 1613 13 Ebert BL, Galili N, Tamayo P, Bosco J, Mak R, Pretz J et al. An erythroid differ- malignancies without del[5q] treated with lenalidomide. J Hematol Oncol 2012; entiation signature predicts response to lenalidomide in myelodysplastic syn- 5:4. drome. PLoS Med 2008; 5: e35. 15 Wei S, Chen X, McGraw K, Zhang L, Komrokji R, Clark J et al. Lenalidomide pro- 14 Sugimoto Y, Sekeres MA, Makishima H, Traina F, Visconte V, Jankowska A et al. motes p53 degradation by inhibiting MDM2 auto-ubiquitination in myelodys- Cytogenetic and molecular predictors of response in patients with myeloid plastic syndrome with chromosome 5q deletion. Oncogene 2013; 32: 1110–1120. Loss of Tcf 7 diminishes hematopoietic stem/progenitor cell function Leukemia (2013) 27, 1613–1614; doi:10.1038/leu.2012.354 BMC/HL %LSK The canonical Wnt-b-catenin pathway is an evolutionarily WT 5.0x107 0.24 conserved and tightly regulated pathway in development. Activation of this pathway occurs upon binding of a soluble Wnt Tcf7-/- 4.9x107 0.25 protein to a membrane-associated receptor, and leads to the disruption and inhibition of a protein complex responsible for the 10000 phosphorylation and breakdown of b-catenin. Inhibition of this so- WT called destruction complex, composed of the tumor suppressor Tcf7-/- Apc, the Ser-Thr kinases Gsk-3b and CK-I, and the scaffold and 1000 tumor suppressor protein Axin, results in stabilization and BM cells (nuclear) accumulation of b-catenin. Stabilized b-catenin forms a 6 bipartite transcription factor with the Tcf-Lef family of transcrip- 100 tion factors (including Tcf7, Tcf7l1, Tcf7l2 and Lef1) to activate a Wnt-controlled gene expression program.1 In the hematopoietic system, a role for Wnt signaling was 10 2 first demonstrated during T-cell development in the thymus.
    [Show full text]
  • Catenin Signaling Regulate External Genitalia Formation As an Appendic
    RESEARCH ARTICLE 3969 Development 136, 3969-3978 (2009) doi:10.1242/dev.039438 Dosage-dependent hedgehog signals integrated with Wnt/-catenin signaling regulate external genitalia formation as an appendicular program Shinichi Miyagawa1,2, Anne Moon3,*, Ryuma Haraguchi2,*, Chie Inoue4, Masayo Harada1, Chiaki Nakahara4, Kentaro Suzuki1, Daisuke Matsumaru4, Takehito Kaneko2, Isao Matsuo5, Lei Yang6, Makoto M. Taketo7, Taisen Iguchi8, Sylvia M. Evans9 and Gen Yamada1,4,† Embryonic appendicular structures, such as the limb buds and the developing external genitalia, are suitable models with which to analyze the reciprocal interactions of growth factors in the regulation of outgrowth. Although several studies have evaluated the individual functions of different growth factors in appendicular growth, the coordinated function and integration of input from multiple signaling cascades is poorly understood. We demonstrate that a novel signaling cascade governs formation of the embryonic external genitalia [genital tubercle (GT)]. We show that the dosage of Shh signal is tightly associated with subsequent levels of Wnt/-catenin activity and the extent of external genitalia outgrowth. In Shh-null mouse embryos, both expression of Wnt ligands and Wnt/-catenin signaling activity are downregulated. -catenin gain-of-function mutation rescues defective GT outgrowth and Fgf8 expression in Shh-null embryos. These data indicate that Wnt/-catenin signaling in the distal urethral epithelium acts downstream of Shh signaling during GT outgrowth. The current data also suggest that Wnt/-catenin regulates Fgf8 expression via Lef/Tcf binding sites in a 3Ј conserved enhancer. Fgf8 induces phosphorylation of Erk1/2 and cell proliferation in the GT mesenchyme in vitro, yet Fgf4/8 compound-mutant phenotypes indicate dispensable functions of Fgf4/8 and the possibility of redundancy among multiple Fgfs in GT development.
    [Show full text]
  • Antibodies Against the Clock Proteins Period and Cryptochrome Reveal the Neuronal Organization of the Circadian Clock in the Pea Aphid
    ORIGINAL RESEARCH published: 02 July 2021 doi: 10.3389/fphys.2021.705048 Antibodies Against the Clock Proteins Period and Cryptochrome Reveal the Neuronal Organization of the Circadian Clock in the Pea Aphid Francesca Sara Colizzi 1, Katharina Beer 1, Paolo Cuti 2, Peter Deppisch 1, David Martínez Torres 2, Taishi Yoshii 3 and Charlotte Helfrich-Förster 1* 1Neurobiology and Genetics, Theodor-Boveri-Institute, Biocenter, University of Würzburg, Würzburg, Germany, 2Institute for Integrative Systems Biology (I2SysBio), University of Valencia and CSIC, Valencia, Spain, 3Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan Circadian clocks prepare the organism to cyclic environmental changes in light, temperature, Edited by: or food availability. Here, we characterized the master clock in the brain of a strongly Joanna C. Chiu, photoperiodic insect, the aphid Acyrthosiphon pisum, immunohistochemically with antibodies University of California, Davis, United States against A. pisum Period (PER), Drosophila melanogaster Cryptochrome (CRY1), and crab Reviewed by: Pigment-Dispersing Hormone (PDH). The latter antibody detects all so far known PDHs and Hideharu Numata, PDFs (Pigment-Dispersing Factors), which play a dominant role in the circadian system of Kyoto University, Japan many arthropods. We found that, under long days, PER and CRY are expressed in a rhythmic Annika Fitzpatrick Barber, Rutgers, The State University of manner in three regions of the brain: the dorsal and lateral protocerebrum and the lamina. No New Jersey, United States staining was detected with anti-PDH, suggesting that aphids lack PDF. All the CRY1-positive *Correspondence: cells co-expressed PER and showed daily PER/CRY1 oscillations of high amplitude, while Charlotte Helfrich-Förster charlotte.foerster@biozentrum.
    [Show full text]
  • A Polymorphism in the Human Timeless Gene Is Not Associated
    Sleep Research Online 3(2): 73-76, 2000 1096-214X http://www.sro.org/2000/Pedrazzoli/73/ © 2000 WebSciences Printed in the USA. All rights reserved. A Polymorphism in the Human Timeless Gene is not Associated with Diurnal Preferences In Normal Adults Mario Pedrazzoli1, Lin Ling1, Laurel Finn2, Terry Young2, Daniel Katzenberg1 and Emmanuel Mignot1 1Center for Narco l e p s y , Department of Psychiatry, Stanford University 2De p a r tment of Preventive Medicine, University of Wisconsin-Madison The effect of a single nucleotide polymorphism, a glutamine to arginine amino acid substitution in the human Timeless gene (Q831R, A2634G), on diurnal preferences was studied in a random sample of normal volunteers enrolled in a population- based epidemiology study of the natural history of sleep disorders. We genotyped 528 subjects for this single nucleotide polymorphism and determined morningness-eveningness tendencies using the Horne-Ostberg questionnaire. Our results indicate that Q831R Timeless has no influence on morningness-eveningness tendencies in humans. CURRENT CLAIM: The A to G polymorphism in the position 2634 of human timeless gene is not related with diurnal preferences. Genetic studies in the last years have advanced our mutations. In animals, mutations such as Clock or Tau (recently understanding of the molecular mechanisms responsible for the characterized as the CKe gene, Lowrey et al., 2000) are control of circadian rhythms. Most of these studies have used associated not only with a long or short free running period but model organisms such as Neurospora, Arabidopis, Drosophila also with alterations in the phase angle of entrainment under and mouse.
    [Show full text]
  • So Here's a Figure of This, Here's the Per Gene, Here's Its Promoter
    So here's a figure of this, here's the per gene, here's its promoter. There's a ribosome, and this gene is now active, illustrated by this glow and the gene is producing messenger RNA which is being turned into protein, into period protein by the protein synthesis machinery. Some of those protein molecules are unstable and they are degraded by the cellular machinery, the pink ones. And some of them are stable for reasons, which we will come to tomorrow, and the stable proteins accumulate. And this protein build-up continues, the gene is active, RNA is made, protein is produced, and at some point in the middle of the night there's enough protein which has been produced, and that protein migrates into the nucleus and the protein then acts as a repressor to turn off its own gene expression. And in the morning, when the sun comes up these protein molecules start to turn over, they degrade and disappear over the course of several hours leading to the turn-on of the gene, which begins the next cycle, the next production of RNA. Now this animation is similar to the one I showed you yesterday except now we have the positive transcription factor CYC and CLOCK, which actually bind to the per promoter at this e-box and drive transcription, turning on RNA synthesis and here is the production of the per protein by the ribosome, the unstable, pink proteins, which are rapidly degraded and then every other protein or so, molecule is stabilized and accumulates in the cytoplasm during the evening.
    [Show full text]