DOCSLIB.ORG

Insights Into the Mechanism of Bovine Spermiogenesis Based on Comparative Transcriptomic Studies

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Insights Into the Mechanism of Bovine Spermiogenesis Based on Comparative Transcriptomic Studies bioRxiv preprint doi: https://doi.org/10.1101/2020.09.25.313908; this version posted September 25, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Insights into the mechanism of bovine spermiogenesis based on comparative transcriptomic studies Xin Li 1, Chenying Duan 2, Ruyi Li 2, Dong Wang 1,* 1 Institute of Animal Science, Chinese Academy of Agricultural Sciences, 100193 Beijing, China 2 College of Animal Science and Technology, Jilin Agricultural University, 130118 Changchun, China * Corresponding author: Dong Wang E-mail: [email protected] Keywords: Spermiogenesis, Differentially expressed genes, Homology trends analysis, Protein-regulating network, ADP-ribosyltransferase 3 (ART3) 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.09.25.313908; this version posted September 25, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. (2), resulting in a tremendous waste. At the same Abstract time, about 15% of the couples of childbearing age worldwide are affected by infertility, of To reduce the reproductive loss caused by which 50% are due to male factors (3), and even semen quality and provide theoretical guidance sperm with normal morphology can cause for the eradication of human male infertility, infertility (4). Therefore, it is necessary to differential analysis of the bovine transcriptome explore the mechanism of spermiogenesis, among round spermatids, elongated spermatids, identify important regulatory pathways and and epididymal sperm was carried out with the important functional genes to improve the semen reference of the mouse transcriptome, and the quality and yield of herd sire, and overcome the homology trends of gene expression to the problem of male infertility (2,5). mouse were also analysed. First, to explore the Since histones are gradually replaced by physiological mechanism of spermiogenesis protamine in the process of spermiogenesis, that profoundly affects semen quality, chromosomes are highly condensed, resulting in homological trends of differential genes were the overall shrinkage of the nucleus. At the same compared during spermiogenesis in dairy cattle time, the spermatid gradually polarises, and the and mice. Next, the Gene ontology (GO), Golgi bodies progressively aggregate and transfer Kyoto Encyclopedia of Genes and Genomes to one end of the spermatid, and finally become (KEGG) pathway enrichment, protein-protein specialised into the acrosome, forming the sperm interaction network (PPI network), and head together with the nucleus. The centrioles bioinformatics analysis uncovered the move in the direction opposite to the head and regulation network of acrosome formation extend outward to become the skeleton of the during the transition from round to elongated sperm tail, while mitochondria gradually spermatids. In addition, processes that regulate specialise to form a ring-shaped mitochondrial gene expression during spermiogenesis from sheath, which is attached to the outside of the elongated spermatid to epididymal sperm, such axon filament in the midpiece of the sperm tail. as ubiquitination, acetylation, deacetylation, At the same time, the cytoplasmic residue glycosylation, and the functional gene ART3 gradually fell off. Following these changes, the may play an important role during round spermatid eventually develops into the spermiogenesis. Therefore, its localisation in tadpole-like sperm with head and tail, which are the seminiferous tubule was investigated by mature and stored for a short time in the immunofluorescent analysis, and its structure epididymis (Figure 1). and function were also predicted. This study However, abnormal expression of some provides important data for revealing the important functional genes during mystery of life during spermiogenesis resulting spermiogenesis affects the morphology (6) and from acrosome formation, histone replacement, structure of the sperm, endanger the function of and the fine regulation of gene expression. the sperm, and lead to male sterility (7). The quality standard for frozen bovine semen (GB4143-2008) stipulates that the proportion of Introduction morphologically abnormal spermatozoa in qualified bovine semen should be less than 18%, With the extensive application of artificial while the Real-World Health Organization insemination, breeders have progressively Human Semen Analysis Laboratory Technical focused on semen requirements for the quality Manual (Fifth Edition) stipulates that the and quantity of excellent bulls. Approximately 10 proportion of morphologically abnormal to 60 billion sperms can be obtained from bulls spermatozoa in human semen should be less than when the semen is collected at a frequency of 4%. Therefore, understanding the regulatory three times a week (1). Considering the presence mechanism of spermiogenesis and mining of 20 million sperms per straw and a calving rate important functional genes have become the of 53% (2), each bull can theoretically breed focus of exploration, prevention, and control of about 13,000 to 78,000 offspring yearly. the causes of male breeding difficulty (8,9). However, due to semen quality issuses caused by Therefore, we performed transcriptome insufficient spermiogenesis, the pregnancy rate of analysis of round spermatids, elongated each straw of frozen semen varied by about 20% 2 bioRxiv preprint doi: https://doi.org/10.1101/2020.09.25.313908; this version posted September 25, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. spermatids, and epididymal sperms of cattle. In samples R1, R2, R3, elongated spermatid order to identify differentially expressed genes samples E1, E2, E3, and epididymal sperm that play an important role in spermiogenesis, the samples M1, M2, and M3 were clustered together. same stage mouse transcriptome sequencing data The PCA results (figure 2B) showed that the sum were introduced as a reference. Homology of PC1 (83.9%) and PC2 (12.9%) of mice was comparison analysis between species was used to 96.8%, among which round spermatid samples obtain bovine genes with the same expression SRR3395024, SRR3395025, and SRR3395026, tendency in mice. Through PPI network, GO and elongated spermatid samples SRR3395030, KEGG pathway enrichment, and bioinformatic SRR3395031, SRR3395032, and epididymal analysis of these genes, we initially obtained the sperm samples SRR4423201, SRR4423202, and protein interaction networks and pathways SRR4423204 were clustered together. related to spermiogenesis, such as acrosomal and This showed that the transcripts of various protamine replacement histones. At the same types of spermatids from bulls and mice showed time, the structure and function of the important high intra-group repeatability, and the differences functional gene ART3 were predicted, and the between the groups were large, so that the localisation of the ART3 protein in the samples were representative. seminiferous tubules was analysed by immunofluorescence. These results provide quantitative real-time PCR (qPCR) validation important clues and a theoretical basis for further of the bull RNA-Seq results exploration of the spermiogenesis mechanism, The relative expression of 11 randomly and play a positive role in improving the selected genes was higher in round and elongated reproductive potential of cattle and promoting the spermatids than in sperm, which was consistent exploration of human infertility mechanisms. with the variations from RNA-seq analysis (Fig.3). Among these genes, the relative Results expression levels of LMTK2 and MIGA2 showed no significant difference (p >0.05) in the sperm Analysis of sequencing results of bull deformation stage; the relative expression level of spermatids and sperm DNAL1 was significantly reduced from round After the quality control of the original spermatid to elongated spermatid (p ＜ 0.05); sequencing data, the number of clean reads for ART3, HIP1, SDHA, and YBX2 showed each sample was more than 66.61 million, which significant reduction in relative expression levels accounted for more than 76.7% of the original from elongated spermatid to epididymal sequencing data, and the Clean Q30 Bases ratio spermatid (p <0.05). The relative expression of each sample was higher than 91%, which levels of TEKT2 and PRKAR1A were showed that the sequencing data could be used in significantly reduced from both round spermatid subsequent experiments (Table 1). After quality to elongated spermatid and elongated spermatid control, the data were compared with the bovine to epididymal sperm (p <0.05); the relative reference genome, the alignment ratio of clean expression level of OAZ3 was significantly reads was higher than 86% for each sample increased from round to elongated spermatid (p (Table 1), showing that the sequencing results <0.05), and a significant decline from elongated could cover most of the reference genome, and spermatid to epididymal sperm (p <0.01). The further analyses could reveal the biological qPCR results validated the RNA-Seq results, information
Load more

Recommended publications
  • Hyaluronidase PH20 (SPAM1) Rabbit Polyclonal Antibody – TA337855
    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 TA337855 Hyaluronidase PH20 (SPAM1) Rabbit Polyclonal Antibody Product data: Product Type: Primary Antibodies Applications: WB Recommended Dilution: WB Reactivity: Human Host: Rabbit Isotype: IgG Clonality: Polyclonal Immunogen: The immunogen for anti-SPAM1 antibody is: synthetic peptide directed towards the C- terminal region of Human SPAM1. Synthetic peptide located within the following region: CYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASP Formulation: Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose. Note that this product is shipped as lyophilized powder to China customers. Purification: Affinity Purified Conjugation: Unconjugated Storage: Store at -20°C as received. Stability: Stable for 12 months from date of receipt. Predicted Protein Size: 58 kDa Gene Name: sperm adhesion molecule 1 Database Link: NP_694859 Entrez Gene 6677 Human P38567 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 / 3 Hyaluronidase PH20 (SPAM1) Rabbit Polyclonal Antibody – TA337855 Background: Hyaluronidase degrades hyaluronic acid, a major structural proteoglycan found in extracellular matrices and basement membranes. Six members of the hyaluronidase family are clustered into two tightly linked groups on chromosome 3p21.3 and 7q31.3. This gene was previously referred to as HYAL1 and HYA1 and has since been assigned the official symbol SPAM1; another family member on chromosome 3p21.3 has been assigned HYAL1.
    [Show full text]
  • Genome-Wide Analysis Reveals Selection Signatures Involved in Meat Traits and Local Adaptation in Semi-Feral Maremmana Cattle
    Genome-Wide Analysis Reveals Selection Signatures Involved in Meat Traits and Local Adaptation in Semi-Feral Maremmana Cattle Slim Ben-Jemaa, Gabriele Senczuk, Elena Ciani, Roberta Ciampolini, Gennaro Catillo, Mekki Boussaha, Fabio Pilla, Baldassare Portolano, Salvatore Mastrangelo To cite this version: Slim Ben-Jemaa, Gabriele Senczuk, Elena Ciani, Roberta Ciampolini, Gennaro Catillo, et al.. Genome-Wide Analysis Reveals Selection Signatures Involved in Meat Traits and Local Adaptation in Semi-Feral Maremmana Cattle. Frontiers in Genetics, Frontiers, 2021, 10.3389/fgene.2021.675569. hal-03210766 HAL Id: hal-03210766 https://hal.inrae.fr/hal-03210766 Submitted on 28 Apr 2021 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution| 4.0 International License ORIGINAL RESEARCH published: 28 April 2021 doi: 10.3389/fgene.2021.675569 Genome-Wide Analysis Reveals Selection Signatures Involved in Meat Traits and Local Adaptation in Semi-Feral Maremmana Cattle Slim Ben-Jemaa 1, Gabriele Senczuk 2, Elena Ciani 3, Roberta
    [Show full text]
  • Program Nr: 1 from the 2004 ASHG Annual Meeting Mutations in A
    Program Nr: 1 from the 2004 ASHG Annual Meeting Mutations in a novel member of the chromodomain gene family cause CHARGE syndrome. L.E.L.M. Vissers1, C.M.A. van Ravenswaaij1, R. Admiraal2, J.A. Hurst3, B.B.A. de Vries1, I.M. Janssen1, W.A. van der Vliet1, E.H.L.P.G. Huys1, P.J. de Jong4, B.C.J. Hamel1, E.F.P.M. Schoenmakers1, H.G. Brunner1, A. Geurts van Kessel1, J.A. Veltman1. 1) Dept Human Genetics, UMC Nijmegen, Nijmegen, Netherlands; 2) Dept Otorhinolaryngology, UMC Nijmegen, Nijmegen, Netherlands; 3) Dept Clinical Genetics, The Churchill Hospital, Oxford, United Kingdom; 4) Children's Hospital Oakland Research Institute, BACPAC Resources, Oakland, CA. CHARGE association denotes the non-random occurrence of ocular coloboma, heart defects, choanal atresia, retarded growth and development, genital hypoplasia, ear anomalies and deafness (OMIM #214800). Almost all patients with CHARGE association are sporadic and its cause was unknown. We and others hypothesized that CHARGE association is due to a genomic microdeletion or to a mutation in a gene affecting early embryonic development. In this study array- based comparative genomic hybridization (array CGH) was used to screen patients with CHARGE association for submicroscopic DNA copy number alterations. De novo overlapping microdeletions in 8q12 were identified in two patients on a genome-wide 1 Mb resolution BAC array. A 2.3 Mb region of deletion overlap was defined using a tiling resolution chromosome 8 microarray. Sequence analysis of genes residing within this critical region revealed mutations in the CHD7 gene in 10 of the 17 CHARGE patients without microdeletions, including 7 heterozygous stop-codon mutations.
    [Show full text]
  • Mammalian Male Germ Cells Are Fertile Ground for Expression Profiling Of
    REPRODUCTIONREVIEW Mammalian male germ cells are fertile ground for expression profiling of sexual reproduction Gunnar Wrobel and Michael Primig Biozentrum and Swiss Institute of Bioinformatics, Klingelbergstrasse 50-70, 4056 Basel, Switzerland Correspondence should be addressed to Michael Primig; Email: [email protected] Abstract Recent large-scale transcriptional profiling experiments of mammalian spermatogenesis using rodent model systems and different types of microarrays have yielded insight into the expression program of male germ cells. These studies revealed that an astonishingly large number of loci are differentially expressed during spermatogenesis. Among them are several hundred transcripts that appear to be specific for meiotic and post-meiotic germ cells. This group includes many genes that were pre- viously implicated in spermatogenesis and/or fertility and others that are as yet poorly characterized. Profiling experiments thus reveal candidates for regulation of spermatogenesis and fertility as well as targets for innovative contraceptives that act on gene products absent in somatic tissues. In this review, consolidated high density oligonucleotide microarray data from rodent total testis and purified germ cell samples are analyzed and their impact on our understanding of the transcriptional program governing male germ cell differentiation is discussed. Reproduction (2005) 129 1–7 Introduction 2002, Sadate-Ngatchou et al. 2003) and the absence of cAMP responsive-element modulator (Crem) and deleted During mammalian male
    [Show full text]
  • Sperm Proteins SOF1, TMEM95, and SPACA6 Are Required for Sperm−Oocyte Fusion in Mice
    Sperm proteins SOF1, TMEM95, and SPACA6 are required for sperm−oocyte fusion in mice Taichi Nodaa,1, Yonggang Lua,1, Yoshitaka Fujiharaa,2, Seiya Ouraa,b, Takayuki Koyanoc, Sumire Kobayashia,b, Martin M. Matzukd,e,3, and Masahito Ikawaa,f,3 aResearch Institute for Microbial Diseases, Osaka University, 565-0871 Osaka, Japan; bGraduate School of Pharmaceutical Sciences, Osaka University, 565-0871 Osaka, Japan; cDivision of Molecular Genetics, Shigei Medical Research Institute, 701-0202 Okayama, Japan; dCenter for Drug Discovery, Baylor College of Medicine, Houston, TX 77030; eDepartment of Pathology & Immunology, Baylor College of Medicine, Houston, TX 77030; and fThe Institute of Medical Science, The University of Tokyo, 108-8639 Tokyo, Japan Contributed by Martin M. Matzuk, March 19, 2020 (sent for review December 27, 2019; reviewed by Matteo Avella and Andrea Pauli) Sperm−oocyte membrane fusion is one of the most important protein, JUNO (also known as IZUMO1 receptor [IZUMO1R] events for fertilization. So far, IZUMO1 and Fertilization Influenc- and folate receptor 4 [FOLR4]) as an IZUMO1 receptor on the ing Membrane Protein (FIMP) on the sperm membrane and CD9 oocyte plasma membrane. IZUMO1 and JUNO form a 1:1 and JUNO (IZUMO1R/FOLR4) on the oocyte membrane have been complex (12), and critical residues to form this interaction were identified as fusion-required proteins. However, the molecular identified by X-ray crystal structure analysis (13–15). However, mechanisms for sperm−oocyte fusion are still unclear. Here, we in vitro studies implied that IZUMO1 may be responsible for show that testis-enriched genes, sperm−oocyte fusion required 1 sperm−oocyte membrane adhesion instead of fusion (16, 17).
    [Show full text]
  • TMEM95 Is a Sperm Membrane Protein Essential for Mammalian
    RESEARCH ARTICLE TMEM95 is a sperm membrane protein essential for mammalian fertilization Ismael Lamas-Toranzo1†, Julieta G Hamze2†, Enrica Bianchi3, Beatriz Ferna´ ndez-Fuertes4,5, Serafı´nPe´ rez-Cerezales1, Ricardo Laguna-Barraza1, Rau´l Ferna´ ndez-Gonza´ lez1, Pat Lonergan4, Alfonso Gutie´ rrez-Ada´ n1, Gavin J Wright3, Marı´aJime´ nez-Movilla2*, Pablo Bermejo-A´ lvarez1* 1Animal Reproduction Department, INIA, Madrid, Spain; 2Department of Cell Biology and Histology, Medical School, University of Murcia, IMIB-Arrixaca, Murcia, Spain; 3Cell Surface Signalling Laboratory, Wellcome Trust Sanger Institute, Cambridge, United Kingdom; 4School of Agriculture and Food Science, University College Dublin, Dublin, Ireland; 5Department of Biology, Faculty of Sciences, Institute of Food and Agricultural Technology, University of Girona, Girona, Spain Abstract The fusion of gamete membranes during fertilization is an essential process for sexual reproduction. Despite its importance, only three proteins are known to be indispensable for sperm- egg membrane fusion: the sperm proteins IZUMO1 and SPACA6, and the egg protein JUNO. Here we demonstrate that another sperm protein, TMEM95, is necessary for sperm-egg interaction. TMEM95 ablation in mice caused complete male-specific infertility. Sperm lacking this protein were morphologically normal exhibited normal motility, and could penetrate the zona pellucida and bind to the oolemma. However, once bound to the oolemma, TMEM95-deficient sperm were unable to fuse with the egg membrane or penetrate into the ooplasm, and fertilization could only be achieved by mechanical injection of one sperm into the ooplasm, thereby bypassing membrane *For correspondence: fusion. These data demonstrate that TMEM95 is essential for mammalian fertilization. [email protected] (Mı´Je´-M); [email protected] (PB-A´ ) †These authors contributed equally to this work Introduction In sexually reproducing species, life begins with the fusion of two gametes during fertilization.
    [Show full text]
  • Gene Section Mini Review
    Atlas of Genetics and Cytogenetics in Oncology and Haematology OPEN ACCESS JOURNAL AT INIST-CNRS Gene Section Mini Review SPAM1 (sperm adhesion molecule 1 (PH -20 hyaluronidase, zona pellucida binding)) Asli Sade, Sreeparna Banerjee Department of Biology, Middle East Technical University, Ankara 06531, Turkey (AS, SB) Published in Atlas Database: March 2010 Online updated version : http://AtlasGeneticsOncology.org/Genes/SPAM1ID42361ch7q31.html DOI: 10.4267/2042/44921 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2010 Atlas of Genetics and Cytogenetics in Oncology and Haematology are clustered on chromosome 3p21.3 and the other Identity three (HYAL4, SPAM1 and HYALP1) are clustered on Other names: EC 3.2.1.35, HYA1, HYAL1, HYAL3, chromosome 7q31.3. Of the three genes on HYAL5, Hyal-PH20, MGC26532, PH-20, PH20, chromosome 7q31.3, HYALP1 is an expressed SPAG15 pseudogene. The extensive homology between the six HGNC (Hugo): SPAM1 hyaluronidase genes suggests an ancient gene duplication event before the emergence of modern Location: 7q31.32 mammals. Local order: According to NCBI Map Viewer, genes flanking SPAM1 in centromere to telomere direction Description on 7q31.3 are: According to Entrez Gene, SPAM1 gene maps to locus - HYALP1 7q31.3 hyaluronoglucosaminidase NC_000007 and spans a region of 46136 bp. According pseudogene 1 to Spidey (mRNA to genomic sequence alignment - HYAL4 7q31.3 hyaluronoglucosaminidase 4 tool), SPAM1 has 7 exons, the sizes being 78, 112, - SPAM1 7q31.3 sperm adhesion molecule 1 1160, 90, 441, 99 and 404. - TMEM229A 7q31.32 transmembrane protein 229A - hCG_1651160 7q31.33 SSU72 RNA polymerase II Transcription CTD phosphatase homolog pseudogene The SPAM1 mRNA has two isoforms; transcript Note: SPAM1 is a glycosyl-phosphatidyl inositol variant 1 (NM_003117) a 2395 bp mRNA and (GPI)-anchored enzyme found in all mammalian transcript variant 2 (NM_153189) a 2009 bp mRNA.
    [Show full text]
  • Binding of Sperm Protein Izumo1 and Its Egg Receptor Juno Drives Cd9
    © 2014. Published by The Company of Biologists Ltd | Development (2014) 141, 3732-3739 doi:10.1242/dev.111534 RESEARCH ARTICLE Binding of sperm protein Izumo1 and its egg receptor Juno drives Cd9 accumulation in the intercellular contact area prior to fusion during mammalian fertilization Myriam Chalbi1, Virginie Barraud-Lange2, Benjamin Ravaux1, Kevin Howan1, Nicolas Rodriguez3, Pierre Soule3, Arnaud Ndzoudi2, Claude Boucheix4,5, Eric Rubinstein4,5, Jean Philippe Wolf2, Ahmed Ziyyat2, Eric Perez1, Frédéric Pincet1 and Christine Gourier1,* ABSTRACT 2006), have so far been shown to be essential. These three Little is known about the molecular mechanisms that induce gamete molecules are also present on human egg and sperm cells. Izumo1 fusion during mammalian fertilization. After initial contact, adhesion is a testis immunoglobulin superfamily type 1 (IgSF) protein, between gametes only leads to fusion in the presence of three expressed at the plasma membrane of acrosome-reacted sperm (Satouh et al., 2012), and is highly conserved in mammals (Grayson membrane proteins that are necessary, but insufficient, for fusion: Izumo1 Izumo1 on sperm, its receptor Juno on egg and Cd9 on egg. What and Civetta, 2012). Female mice deleted for the gene have happens during this adhesion phase is a crucial issue. Here, we normal fertility, but males are completely sterile despite normal mating behavior and normal sperm production (Inoue et al., 2005). demonstrate that the intercellular adhesion that Izumo1 creates with Cd9−/− Juno is conserved in mouse and human eggs. We show that, along Similar features are observed for Cd9 on egg cells. female are healthy but severely subfertile because of defective sperm-egg with Izumo1, egg Cd9 concomitantly accumulates in the adhesion in vivo area.
    [Show full text]
  • Hippo and Sonic Hedgehog Signalling Pathway Modulation of Human Urothelial Tissue Homeostasis
    Hippo and Sonic Hedgehog signalling pathway modulation of human urothelial tissue homeostasis Thomas Crighton PhD University of York Department of Biology November 2020 Abstract The urinary tract is lined by a barrier-forming, mitotically-quiescent urothelium, which retains the ability to regenerate following injury. Regulation of tissue homeostasis by Hippo and Sonic Hedgehog signalling has previously been implicated in various mammalian epithelia, but limited evidence exists as to their role in adult human urothelial physiology. Focussing on the Hippo pathway, the aims of this thesis were to characterise expression of said pathways in urothelium, determine what role the pathways have in regulating urothelial phenotype, and investigate whether the pathways are implicated in muscle-invasive bladder cancer (MIBC). These aims were assessed using a cell culture paradigm of Normal Human Urothelial (NHU) cells that can be manipulated in vitro to represent different differentiated phenotypes, alongside MIBC cell lines and The Cancer Genome Atlas resource. Transcriptomic analysis of NHU cells identified a significant induction of VGLL1, a poorly understood regulator of Hippo signalling, in differentiated cells. Activation of upstream transcription factors PPARγ and GATA3 and/or blockade of active EGFR/RAS/RAF/MEK/ERK signalling were identified as mechanisms which induce VGLL1 expression in NHU cells. Ectopic overexpression of VGLL1 in undifferentiated NHU cells and MIBC cell line T24 resulted in significantly reduced proliferation. Conversely, knockdown of VGLL1 in differentiated NHU cells significantly reduced barrier tightness in an unwounded state, while inhibiting regeneration and increasing cell cycle activation in scratch-wounded cultures. A signalling pathway previously observed to be inhibited by VGLL1 function, YAP/TAZ, was unaffected by VGLL1 manipulation.
    [Show full text]
  • Supp Table 6.Pdf
    Supplementary Table 6. Processes associated to the 2037 SCL candidate target genes ID Symbol Entrez Gene Name Process NM_178114 AMIGO2 adhesion molecule with Ig-like domain 2 adhesion NM_033474 ARVCF armadillo repeat gene deletes in velocardiofacial syndrome adhesion NM_027060 BTBD9 BTB (POZ) domain containing 9 adhesion NM_001039149 CD226 CD226 molecule adhesion NM_010581 CD47 CD47 molecule adhesion NM_023370 CDH23 cadherin-like 23 adhesion NM_207298 CERCAM cerebral endothelial cell adhesion molecule adhesion NM_021719 CLDN15 claudin 15 adhesion NM_009902 CLDN3 claudin 3 adhesion NM_008779 CNTN3 contactin 3 (plasmacytoma associated) adhesion NM_015734 COL5A1 collagen, type V, alpha 1 adhesion NM_007803 CTTN cortactin adhesion NM_009142 CX3CL1 chemokine (C-X3-C motif) ligand 1 adhesion NM_031174 DSCAM Down syndrome cell adhesion molecule adhesion NM_145158 EMILIN2 elastin microfibril interfacer 2 adhesion NM_001081286 FAT1 FAT tumor suppressor homolog 1 (Drosophila) adhesion NM_001080814 FAT3 FAT tumor suppressor homolog 3 (Drosophila) adhesion NM_153795 FERMT3 fermitin family homolog 3 (Drosophila) adhesion NM_010494 ICAM2 intercellular adhesion molecule 2 adhesion NM_023892 ICAM4 (includes EG:3386) intercellular adhesion molecule 4 (Landsteiner-Wiener blood group)adhesion NM_001001979 MEGF10 multiple EGF-like-domains 10 adhesion NM_172522 MEGF11 multiple EGF-like-domains 11 adhesion NM_010739 MUC13 mucin 13, cell surface associated adhesion NM_013610 NINJ1 ninjurin 1 adhesion NM_016718 NINJ2 ninjurin 2 adhesion NM_172932 NLGN3 neuroligin
    [Show full text]
  • Hyaluronidase PH20 (SPAM1) (NM 003117) Human Mass Spec Standard – PH305378 | Origene
    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 PH305378 Hyaluronidase PH20 (SPAM1) (NM_003117) Human Mass Spec Standard Product data: Product Type: Mass Spec Standards Description: SPAM1 MS Standard C13 and N15-labeled recombinant protein (NP_003108) Species: Human Expression Host: HEK293 Expression cDNA Clone RC205378 or AA Sequence: Predicted MW: 58.2 kDa Protein Sequence: >RC205378 representing NM_003117 Red=Cloning site Green=Tags(s) MGVLKFKHIFFRSFVKSSGVSQIVFTFLLIPCCLTLNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDM SLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNL GMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLL RPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRN RVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSIMRSMKS CLLLDNYMETILNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGK PTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLS ATMFIWRLEVWDQGISRIGFF TRTRPLEQKLISEEDLAANDILDYKDDDDKV Tag: C-Myc/DDK Purity: > 80% as determined by SDS-PAGE and Coomassie blue staining Concentration: 50 ug/ml as determined by BCA Labeling Method: Labeled with [U- 13C6, 15N4]-L-Arginine and [U- 13C6, 15N2]-L-Lysine Buffer: 100 mM glycine, 25 mM Tris-HCl, pH 7.3. Store at -80°C. Avoid repeated freeze-thaw cycles. Stable for 3 months from receipt of products under proper storage and handling conditions. RefSeq:
    [Show full text]
  • Integrating Data Mining, Network Pharmacology, and Molecular
    Integrating Data Mining, Network Pharmacology, and Molecular Docking Verication to Investigate the Molecular Mechanism of Traditional Chinese Medicine Prescriptions for Treating Male Infertility Xue Bai Beijing University of Chinese Medicine Yibo Tang Beijing University of Chinese Medicine Qiang Li Beijing University of Chinese Medicine Guimin Liu Beijing University of Chinese Medicine Dan Liu Beijing University of Chinese Medicine Xiaolei Fan Beijing University of Chinese Medicine Zhejun Liu Beijing University of Chinese Medicine Shujun Yu Beijing University of Chinese Medicine Tian Tang Beijing University of Chinese Medicine Shuyan Wang Beijing University of Chinese Medicine Lingru Li Beijing University of Chinese Medicine Kailin Zhou Beijing University of Chinese Medicine Yanfei Zheng Beijing University of Chinese Medicine Zhenquan Liu ( [email protected] ) Research Page 1/42 Keywords: Traditional Chinese medicine, male infertility, data mining, network pharmacology, molecular docking Posted Date: March 4th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-264555/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 2/42 Abstract Background: Male infertility (MI) affects almost 5% adult men worldwide, and 75% of these cases are unexplained idiopathic. There are limitations in the current treatment due to the unclear mechanism of MI, which highlight the urgent need for a more effective strategy or drug. Traditional Chinese Medicine (TCM) prescriptions have been used to treat MI for thousands of years, but their molecular mechanism is not well dened. Methods: Aiming at revealing the molecular mechanism of TCM prescriptions on MI, a comprehensive strategy integrating data mining, network pharmacology, and molecular docking verication was performed.
    [Show full text]