DOCSLIB.ORG

Investigating the Role of Ephrins and Their Receptors in Mouse Folliculogenesis and Ovulation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Investigating the Role of Ephrins and Their Receptors in Mouse Folliculogenesis and Ovulation Western University Scholarship@Western Electronic Thesis and Dissertation Repository 4-23-2015 12:00 AM Investigating the role of ephrins and their receptors in mouse folliculogenesis and ovulation Adrian Buensuceso The University of Western Ontario Supervisor Dr. Bonnie Deroo The University of Western Ontario Graduate Program in Biochemistry A thesis submitted in partial fulfillment of the equirr ements for the degree in Doctor of Philosophy © Adrian Buensuceso 2015 Follow this and additional works at: https://ir.lib.uwo.ca/etd Part of the Endocrinology Commons Recommended Citation Buensuceso, Adrian, "Investigating the role of ephrins and their receptors in mouse folliculogenesis and ovulation" (2015). Electronic Thesis and Dissertation Repository. 2780. https://ir.lib.uwo.ca/etd/2780 This Dissertation/Thesis is brought to you for free and open access by Scholarship@Western. It has been accepted for inclusion in Electronic Thesis and Dissertation Repository by an authorized administrator of Scholarship@Western. For more information, please contact [email protected]. INVESTIGATING THE ROLE OF EPHRINS AND THEIR RECEPTORS IN MOUSE FOLLICULOGENESIS AND OVULATION (Thesis format: Integrated-Article) by Adrian Vincent Carlos Buensuceso Graduate Program in Biochemistry A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy The School of Graduate and Postdoctoral Studies The University of Western Ontario London, Ontario, Canada © Adrian Buensuceso 2015 Abstract Follicle stimulating hormone (FSH) promotes granulosa cell (GC) proliferation, differentiation, and steroidogenesis. This series of events is critical for female fertility, and culminates in the formation of mature follicles responsive to the surge of luteinizing hormone (LH) that triggers ovulation. Ephrins ( Efn genes) and Eph receptors ( Eph genes) are membrane-associated signaling molecules that mediate communication at sites of cell-cell contact, and have been extensively studied in the context of embryonic development. The published literature contains several reports of ovarian Eph and Efn expression, although their precise roles in the ovary are unknown. Dysregulation of Efna5 in GCs of the subfertile Esr2 -/- mouse suggests a role for Eph-ephrin signaling in ovarian function. We sought to investigate Eph receptors and ephrins in the mouse ovary using gonadotropin-stimulated animal models and cultured GCs. We identified several Efn and Eph genes for which expression is enhanced in GCs of the gonadotropin-stimulated mouse. Furthermore, we determined that cultured GCs stimulated with recombinant ephrin-A5 or EphA5 exhibit reduced cell spreading or adhesion, respectively, indicating a cell-autonomous response to Eph-ephrin stimulation. In order to ascertain the importance of ephrin-A5 in female fertility, we performed a reproductive assessment of females lacking Efna5 , the sole ephrin-encoding gene upregulated by FSH in GCs. Efna5 -/- females are subfertile and exhibit an impaired response to LH, displaying attenuated ovulatory potential, reduced ovarian expression of Pgr , Ptgs2 , and Adamts4 , as well as abnormal follicle rupture. We also found an increased incidence of multi-oocyte follicles in adult, but not juvenile, Efna5 -/- females, indicating follicle merging. Finally, we determined that the mouse genomic region upstream of Epha5 is transcriptionally activated by cAMP in a protein kinase A-dependent manner. Transcriptional activation of Efna5 , Epha3 , Epha5 , Epha8 , and Ephb2 is reduced in GCs of eCG-treated Esr2 -/- females, which exhibit impaired cAMP production. These results suggest that cAMP is a novel transcriptional regulator for several Eph and Efn genes. Our findings establish a place for ephrins and their receptors within the current model of gonadotropin-dependent follicle growth and ovulation, and identify cAMP as a novel transcriptional regulator of Epha5 and possibly other Efn and Eph genes. ii Keywords Eph receptor, ephrin, mouse, granulosa cell, follicle, ovary, folliculogenesis, ovulation, follicle rupture, gonadotropin iii Co-authorship statement Chapter 2: Buensuceso AV, Deroo BJ. The ephrin signaling pathway regulates morphology and adhesion of mouse granulosa cells in vitro. Biol Reprod . 2013;88(1):25. Dr. Bonnie Deroo conceived the project and co-wrote the manuscript. Chapter 3: Dr. Alexander Son and Dr. Renping Zhou (Rutgers University, NJ, USA) provided Efna5 +/+ and Efna5 -/- ovaries used for multi-oocyte follicle counts as well as founder mice for our mouse colony. Marilène Paquet performed histological examination of Efna5 +/+ and Efna5 -/- ovaries. Benjamin M. Withers performed counts of follicles containing expanded COCs and corpora lutea, and performed histological archiving. Dr. Bonnie Deroo conceived the project and co-wrote the manuscript which was submitted to Endocrinology , ID# EN-15-1216. Chapter 4: Erin Parker conducted genotyping for some Esr2 +/+ and Esr2 -/- mice. Dr. Bonnie Deroo conceived the project. iv Acknowledgements First, I would like to thank my supervisor and mentor, Dr. Bonnie Deroo, for her excellent mentorship. She has guided my development as a scientist, providing me with constructive criticism and encouragement all along the way. She has always made time for me, and has built a research environment where I could flourish. I enjoy science now more than ever, and I credit her with that. I would like to thank my supervisory committee members, Dr. Gabriel DiMattia and Dr. Eric Ball. Their guidance has played a major role in the development of my research. I would also like to acknowledge my colleagues in the Deroo Lab: Caitie O’Flynn, Ola Carrier, and Dr. Macarena Pampillo. I value their insight and advice, and have very much enjoyed working with them. Furthermore, Dr. Gabriel DiMattia, Dr. Trevor Shepherd, Dr. Tom Drysdale, and Dr. Chris Pin, as well as their trainees and technicians, have given me valuable advice during our big group lab meetings. Many thanks as well to Robert Gauthier and Penny Smithers-Fortnum for helping to care for our mice. Lastly, I would like to express my appreciation to my colleagues in the Victoria Research Labs and London Cancer Research Centre: Mateusz Rytelewski, Jessica Tong, Colin Way, Dr. Saman Maleki Vareki, Rene Figueredo, Dr. Julio Masabanda, and Dr. Pete Ferguson. These people have all helped me to grow as a scientist and as a person during my time as a graduate student. v Ethics Approval Experiments were performed in compliance with the guidelines set by the Canadian Council for Animal Care, and the policies and procedures approved by the University of Western Ontario Council on Animal Care (Protocol Number: 2008-042). vi Table of Contents Abstract ............................................................................................................................... ii Keywords ........................................................................................................................... iii Co-authorship statement .................................................................................................... iv Acknowledgements ............................................................................................................. v Ethics Approval ................................................................................................................. vi Table of Contents .............................................................................................................. vii List of Figures .................................................................................................................. xiv List of Tables ................................................................................................................... xvi List of Abbreviations ...................................................................................................... xvii 1 Chapter 1: Introduction ........................................................................................ 1 1.1 Stages of Ovarian Follicle Growth ....................................................................... 1 1.1.1 Introduction to Folliculogenesis ....................................................................1 1.1.2 Formation of primordial follicles ...................................................................1 1.1.3 Follicle-stimulating hormone-independent follicle growth ...........................2 1.1.4 FSH-dependent follicle growth ......................................................................3 1.1.5 LH-dependent oocyte maturation, cumulus expansion, ovulation, and corpus luteum formation ......................................................................................................4 1.2 The hypothalamic-pituitary-gonadal axis........................................................... 10 1.3 Eph receptors and Ephrins.................................................................................. 12 1.3.1 Overview ......................................................................................................12 1.3.2 Ephrins .........................................................................................................14 1.3.3 Eph receptors ...............................................................................................15 1.3.4 Initiation of Eph-Ephrin Signaling...............................................................17 vii 1.3.5 Eph-ephrin Signal Transduction ..................................................................17
Load more

Recommended publications
  • Targeting Synthetic Lethality Between the SRC Kinase and the EPHB6 Receptor May Benefit Cancer Treatment
    www.impactjournals.com/oncotarget/ Oncotarget, Vol. 7, No. 31 Research Paper Targeting synthetic lethality between the SRC kinase and the EPHB6 receptor may benefit cancer treatment James M. Paul1,*, Behzad Toosi2,*, Frederick S. Vizeacoumar2,*, Kalpana Kalyanasundaram Bhanumathy2, Yue Li3,4,5, Courtney Gerger2, Amr El Zawily2,6, Tanya Freywald7, Deborah H. Anderson7, Darrell Mousseau8, Rani Kanthan2, Zhaolei Zhang3,4, Franco J. Vizeacoumar2,7, Andrew Freywald2 1Department of Biochemistry, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada 2Department of Pathology and Laboratory Medicine, College of Medicine, University of Saskatchewan, Royal University Hospital, Saskatoon, SK, S7N 0W8, Canada 3Department of Computer Science, University of Toronto, Toronto, ON, M5S 3G4, Canada 4The Donnelly Centre, University of Toronto, Toronto, ON, M5S 3E1, Canada 5Present address: Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA 6Faculty of Science, Damanhour University, Damanhour, 22516, Egypt 7Cancer Research, Saskatchewan Cancer Agency, Saskatoon, SK, S7N 5E5, Canada 8Cell Signaling Laboratory, Neuroscience Cluster, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada *These authors contributed equally to this work Correspondence to: Franco J. Vizeacoumar, email: [email protected] Andrew Freywald, email: [email protected] Keywords: breast cancer, genetic interaction, synthetic lethality, EPHB6, SRC kinase Received: April 22, 2016 Accepted: June 17, 2016 Published: July 13, 2016 ABSTRACT Application of tumor genome sequencing has identified numerous loss-of-function alterations in cancer cells. While these alterations are difficult to target using direct interventions, they may be attacked with the help of the synthetic lethality (SL) approach. In this approach, inhibition of one gene causes lethality only when another gene is also completely or partially inactivated.
    [Show full text]
  • Increased Expression of Epidermal Growth Factor Receptor and Betacellulin During the Early Stage of Gastric Ulcer Healing
    505-510 11/6/08 12:55 Page 505 MOLECULAR MEDICINE REPORTS 1: 505-510, 2008 505 Increased expression of epidermal growth factor receptor and betacellulin during the early stage of gastric ulcer healing GEUN HAE CHOI, HO SUNG PARK, KYUNG RYOUL KIM, HA NA CHOI, KYU YUN JANG, MYOUNG JA CHUNG, MYOUNG JAE KANG, DONG GEUN LEE and WOO SUNG MOON Department of Pathology, Institute for Medical Sciences, Chonbuk National University Medical School and the Center for Healthcare Technology Development, Jeonju, Korea Received January 2, 2008; Accepted February 22, 2008 Abstract. Epidermal growth factor receptor (EGFR) is from tissue necrosis triggered by mucosal ischemia, free important for the proliferation and differentiation of gastric radical formation and the cessation of nutrient delivery, which mucosal cells. Betacellulin (BTC) is a novel ligand for EGFR are caused by vascular and microvascular injury such as Since their role is unclear in the ulcer healing process, we thrombi, constriction or other occlusions (2). Tissue necrosis investigated their expression. Gastric ulcers in 30 Sprague- and the release of leukotriene B attract leukocytes and Dawley rats were induced by acetic acid. RT-PCR and macrophages, which release pro-inflammatory cytokines Western blotting were performed to detect EGFR and BTC. (e.g. TNFα, IL-1α, and IL-1ß). These in turn activate local Immunohistochemical studies were performed to detect fibroblasts, endothelial and epithelial cells. Histologically, an EGFR, BTC and proliferating cell nuclear antigen (PCNA). ulcer has two characteristic structures: a distinct ulcer margin The expression of EGFR and the BTC gene was significantly formed by the adjacent non-necrotic mucosa, and granulation increased at 12 h, 24 h and 3 days after ulcer induction tissue composed of fibroblasts, macrophages and proliferating (P<0.05).
    [Show full text]
  • Implications for Diabetic Nephropathy
    ORIGINAL ARTICLE Hyperglycemia Causes Renal Cell Damage via CCN2-Induced Activation of the Tropomyosin Receptor Kinase A, TrkA Implications for Diabetic Nephropathy Maria Fragiadaki,1 Nicola Hill,1 Reiko Hewitt,1 George Bou-Gharios,1,2 Terence Cook,1,3 Frederick W. Tam,1 Jan Domin,1,4 and Roger M. Mason1 CCN2, a secreted profibrotic protein, is highly expressed in di- biopsy specimens from patients with different stages of DN abetic nephropathy (DN) and implicated in its pathogenesis; (6). CCN2 is also induced by TGF-b, hypoxia (low oxygen), however, the actions of CCN2 in DN remain elusive. We previously and advanced glycation-end products, all of which are demonstrated that CCN2 triggers signaling via tropomyosin re- stimulants that result in kidney damage (6–9). Moreover, ceptor kinase A (TrkA). Trace expression of TrkA is found in CCN2 overexpression in transgenic mice with streptozotocin- normal kidneys, but its expression is elevated in several nephrop- induced DN led to worsening of the disease, thus indicating athies; yet its role in DN is unexplored. In this study we show de the latter may play a profibrotic role after primary injury in novo expression of TrkA in human and murine DN. We go on to the context of diabetes (10). study the molecular mechanisms leading to TrkA activation and show – More evidence that CCN2 is a profibrotic protein comes that it involves hypoxia, as demonstrated by ischemia reperfusion fi injury and in vitro experiments mimicking hypoxia, implicating from studies of genetically modi ed animals. Sonnylal et al. hypoxia as a common pathway leading to disease.
    [Show full text]
  • Recombinant Human Betacellulin Promotes the Neogenesis of -Cells
    Recombinant Human Betacellulin Promotes the Neogenesis of ␤-Cells and Ameliorates Glucose Intolerance in Mice With Diabetes Induced by Selective Alloxan Perfusion Koji Yamamoto, Jun-ichiro Miyagawa, Masako Waguri, Reiko Sasada, Koichi Igarashi, Ming Li, Takao Nammo, Makoto Moriwaki, Akihisa Imagawa, Kazuya Yamagata, Hiromu Nakajima, Mitsuyoshi Namba, Yoshihiro Tochino, Toshiaki Hanafusa, and Yuji Matsuzawa Betacellulin (BTC), a member of the epidermal growth factor family, is expressed predominantly in the human pancreas and induces the differentiation of a pancreatic ancreatic ␤-cells are thought to be terminally dif- acinar cell line (AR42J) into insulin-secreting cells, ferentiated cells with little ability to regenerate. suggesting that BTC has a physiologically important However, proliferation of preexisting ␤-cells and role in the endocrine pancreas. In this study, we exam- differentiation of ␤-cells from precursor cells, ined the in vivo effect of recombinant human BTC P (rhBTC) on glucose intolerance and pancreatic mor- mainly residing in the pancreatic duct lining, have been phology using a new mouse model with glucose intoler- demonstrated in some animal models (1–5). Recently, we ance induced by selective alloxan perfusion. RhBTC developed a new mouse model of diabetes induced by selec- (1 µg/g body wt) or saline was injected subcutaneously tive perfusion of alloxan (100 µg/g body wt) during the every day from the day after alloxan treatment. The clamping of the superior mesenteric artery (1). In this model, intraperitoneal glucose tolerance test revealed no dif- glucose intolerance spontaneously resolves after one year ference between rhBTC-treated and rhBTC-untreated because of the proliferation of surviving ␤-cells in the non- glucose-intolerant mice at 2–4 weeks.
    [Show full text]
  • Table 2. Significant
    Table 2. Significant (Q < 0.05 and |d | > 0.5) transcripts from the meta-analysis Gene Chr Mb Gene Name Affy ProbeSet cDNA_IDs d HAP/LAP d HAP/LAP d d IS Average d Ztest P values Q-value Symbol ID (study #5) 1 2 STS B2m 2 122 beta-2 microglobulin 1452428_a_at AI848245 1.75334941 4 3.2 4 3.2316485 1.07398E-09 5.69E-08 Man2b1 8 84.4 mannosidase 2, alpha B1 1416340_a_at H4049B01 3.75722111 3.87309653 2.1 1.6 2.84852656 5.32443E-07 1.58E-05 1110032A03Rik 9 50.9 RIKEN cDNA 1110032A03 gene 1417211_a_at H4035E05 4 1.66015788 4 1.7 2.82772795 2.94266E-05 0.000527 NA 9 48.5 --- 1456111_at 3.43701477 1.85785922 4 2 2.8237185 9.97969E-08 3.48E-06 Scn4b 9 45.3 Sodium channel, type IV, beta 1434008_at AI844796 3.79536664 1.63774235 3.3 2.3 2.75319499 1.48057E-08 6.21E-07 polypeptide Gadd45gip1 8 84.1 RIKEN cDNA 2310040G17 gene 1417619_at 4 3.38875643 1.4 2 2.69163229 8.84279E-06 0.0001904 BC056474 15 12.1 Mus musculus cDNA clone 1424117_at H3030A06 3.95752801 2.42838452 1.9 2.2 2.62132809 1.3344E-08 5.66E-07 MGC:67360 IMAGE:6823629, complete cds NA 4 153 guanine nucleotide binding protein, 1454696_at -3.46081884 -4 -1.3 -1.6 -2.6026947 8.58458E-05 0.0012617 beta 1 Gnb1 4 153 guanine nucleotide binding protein, 1417432_a_at H3094D02 -3.13334396 -4 -1.6 -1.7 -2.5946297 1.04542E-05 0.0002202 beta 1 Gadd45gip1 8 84.1 RAD23a homolog (S.
    [Show full text]
  • The Kinase Defective EPHB6 Receptor Tyrosine Kinase Activates MAP Kinase Signaling in Lung Adenocarcinoma
    175-179.qxd 29/5/2009 01:21 ÌÌ ™ÂÏ›‰·175 INTERNATIONAL JOURNAL OF ONCOLOGY 35: 175-179, 2009 175 The kinase defective EPHB6 receptor tyrosine kinase activates MAP kinase signaling in lung adenocarcinoma JUN YU1,2, ETMAR BULK1, PING JI1, ANTJE HASCHER1, STEFFEN KOSCHMIEDER1, WOLFGANG E. BERDEL1 and CARSTEN MÜLLER-TIDOW1 1Department of Medicine, Hematology and Oncology, University of Münster, Münster, Germany; 2Department of Preclinical Experiment Center, Fourth Military Medical University, Xi'an, P.R. China Received January 28, 2009; Accepted March 13, 2009 DOI: 10.3892/ijo_00000326 Abstract. Decreased expression levels of EPHB6, a member Elk-1 (8,9). ERK1/2 are negatively regulated by a family of of the receptor tyrosine kinases (RTKs), are associated with dual-specificity (Thr/Tyr) MAPK phosphatases, known as an increased risk of metastasis development in early stage DUSPs or MKPs, and pharmacologically by MEK inhibitors non-small cell lung cancer (NSCLC). However, the signaling such as U0126 and PD98059 (10). properties of the kinase-defective EPHB6 receptor are not EPH receptors form the largest known subfamily of well-understood. Here, we show that expression of EPHB6 receptor tyrosine kinases, and to date, the EPH subfamily in A549 lung adenocarinoma cells led to phosphorylation of contains 16 members in vertebrates (11,12). The EPH receptors the MAP kinase ERK. Conversely, siRNA based knockdown interact with a family of ligands located on the surfaces of of EPHB6 reversed ERK phosphorylation. Intriguingly, adjacent cells, named Ephrins including Ephrin-As and EPHB6-induced phosphorylation of ERK was uncoupled Ephrin-Bs subgroups. The EPH receptors are also grouped by activation of the Elk-1 transcriptional factor.
    [Show full text]
  • Reproductionresearch
    REPRODUCTIONRESEARCH Bone morphogenetic protein 15 and fibroblast growth factor 10 enhance cumulus expansion, glucose uptake, and expression of genes in the ovulatory cascade during in vitro maturation of bovine cumulus–oocyte complexes Ester S Caixeta, Melanie L Sutton-McDowall1, Robert B Gilchrist1, Jeremy G Thompson1, Christopher A Price2, Mariana F Machado, Paula F Lima and Jose´ Buratini Departamento de Fisiologia, Instituto de Biocieˆncias, Universidade Estadual Paulista, Rubia˜o Junior, Botucatu, Sa˜o Paulo 18618-970, Brazil, 1The Robinson Institute, Research Centre for Reproductive Health, School of Paediatrics and Reproductive Health, The University of Adelaide, Adelaide, South Australia 5005, Australia and 2Centre de Recherche en Reproduction Animale, Faculte´ de Me´decine Ve´te´rinaire, Universite´ de Montre´al, St-Hyacinthe, Quebec, Canada J2S 7C6 Correspondence should be addressed to J Buratini; Email: [email protected] Abstract Oocyte-secreted factors (OSFs) regulate differentiation of cumulus cells and are of pivotal relevance for fertility. Bone morphogenetic protein 15 (BMP15) and fibroblast growth factor 10 (FGF10) are OSFs and enhance oocyte competence by unknown mechanisms. We tested the hypothesis that BMP15 and FGF10, alone or combined in the maturation medium, enhance cumulus expansion and expression of genes in the preovulatory cascade and regulate glucose metabolism favouring hyaluronic acid production in bovine cumulus–oocyte complexes (COCs). BMP15 or FGF10 increased the percentage of fully expanded COCs, but the combination did not further stimulate it. BMP15 increased cumulus cell levels of mRNA encoding a disintegrin and metalloprotease 10 (ADAM10), ADAM17, amphiregulin (AREG), and epiregulin (EREG) at 12 h of culture and of prostaglandin (PG)-endoperoxide synthase 2 (PTGS2), pentraxin 3 (PTX3) and tumor necrosis factor alpha-induced protein 6 (TNFAIP6 (TSG6)) at 22 h of culture.
    [Show full text]
  • Mesenchymal–Epithelial Interactions Involving Epiregulin in Tuberous Sclerosis Complex Hamartomas
    Mesenchymal–epithelial interactions involving epiregulin in tuberous sclerosis complex hamartomas Shaowei Li*, Fumiko Takeuchi*, Ji-an Wang*, Qingyuan Fan*, Toshi Komurasaki†, Eric M. Billings‡, Gustavo Pacheco-Rodriguez‡, Joel Moss‡, and Thomas N. Darling*§ *Department of Dermatology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814-4712; †Molecular Biology Laboratory, Molecular and Pharmacology Laboratories, Taisho Pharmaceutical Co., Ltd., 430-1 Yoshino-cho, Saitma-shi, Saitama 331-9530, Japan; and ‡Translational Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 10, Room 6D05, MSC 1590, Bethesda, MD 20892-1590 Communicated by Martha Vaughan, National Institutes of Health, Bethesda, MD, December 31, 2007 (received for review November 30, 2007) Patients with tuberous sclerosis complex (TSC) develop hamarto- Like other hamartomas, those in TSC skin contain abnormal mas containing biallelic inactivating mutations in either TSC1 or numbers of several types of cells. In the dermis, there are TSC2, resulting in mammalian target of rapamycin (mTOR) activa- increased numbers of large stellate fibroblasts, capillaries, and tion. Hamartomas overgrow epithelial and mesenchymal cells in dermal dendritic cells (6–9). The epidermis is acanthotic (i.e., TSC skin. The pathogenetic mechanisms for these changes had not thickened from increased numbers of keratinocytes in the spi- been investigated, and the existence or location of cells with nous layer). Acanthosis is pronounced in PFs and variable in AFs biallelic mutations (‘‘two-hit’’ cells) was unclear. We compared TSC (7, 8). The epidermis of treated AFs, several months after argon skin hamartomas (angiofibromas and periungual fibromas) with or CO2 laser surgery, no longer appears acanthotic (10, 11).
    [Show full text]
  • Anti-Human EPHA4 / EPH Receptor A4 (Internal) Polyclonal Antibody
    For Research Use Only. REV: 9/9/2014 Anti-Human EPHA4 / EPH Receptor A4 (Internal) Polyclonal Antibody CatalogID MC-2841 Target Protein EPH receptor A4 (EPHA4) Synonyms EPHA4 Antibody, CEK8 Antibody, EK8 Antibody, Ephrin type-A receptor 4 Antibody, HEK8 Antibody, SEK Antibody, TYRO1 Antibody, TYRO1 protein tyrosine kinase Antibody, Tyrosine-protein kinase TYRO1 Antibody, EPH receptor A4 Antibody, EPH-like kinase 8 Antibody Family / Subfamily Protein Kinase / Ephrin Receptor Host EPHA4 antibody was produced in Rabbit Clonality Polyclonal Immunogen Species EPHA4 / EPH Receptor A4 antibody was raised against Human Antigen Type Synthetic peptide Immunogen EPHA4 / EPH Receptor A4 antibody was raised against synthetic 18 amino acid peptide from internal region of human EPHA4. Percent identity with other species by BLAST analysis: Human, Gorilla, Gibbon, Monkey, Marmoset, Panda, Bovine, Dog, Bat, Horse, Pig (100%); Mouse, Rat, Hamster, Elephant, Rabbit, Opossum, Platypus, Xenopus (94%); Turkey, Chicken (89%); Lizard (83%). Specificity Human EPHA4. BLAST analysis of the peptide immunogen showed no homology with other human proteins. Epitope Internal Reactivity Human (human tissues have been tested in IHC with FFPE) Predicted Reactivity Gorilla, Gibbon, Monkey, Bat, Bovine, Dog, Horse, Pig, Mouse, Rat, Hamster, Rabbit, Xenopus Purification Immunoaffinity purified Presentation PBS, 0.1% sodium azide. Recommended Storage Long term: -70°C; Short term: +4°C Uses IHC - Paraffin (3 μg/ml) (Optimal dilution to be determined by the researcher) Size 50 µg Concentration 1 mg/ml For research use only. Not intended for human, diagnostic, therapeutic, or drug use. Immunohistochemistry Anti-EPHA4 antibody MC-2841 IHC of human kidney, glomerulus. Immunohistochemistry of formalin-fixed, paraffin- embedded tissue after heat-induced antigen retrieval.
    [Show full text]
  • Epha3 Inhibits Migration and Invasion of Esophageal Cancer Cells by Activating the Mesenchymal‑Epithelial Transition Process
    722 INTERNATIONAL JOURNAL OF ONCOLOGY 54: 722-732, 2019 EphA3 inhibits migration and invasion of esophageal cancer cells by activating the mesenchymal‑epithelial transition process XIA CHEN1,2, BIN LU2,3, QIAN MA2, CHENG-DONG JI4 and JIAN-ZHONG LI3 1Key Laboratory, Yangpu Hospital, Tongji University School of Medicine, Shanghai 200090; 2International Joint Cancer Institute; 3Department of Biochemical Pharmacy, Second Military Medical University, Shanghai 200433; 4Department of Scientific Research Management, Yangpu Hospital, Tongji University School of Medicine, Shanghai 200090, P.R. China Received June 13, 2018; Accepted November 2, 2018 DOI: 10.3892/ijo.2018.4639 Abstract. Eph receptor tyrosine kinases are critical for cell-cell Introduction communication during normal and oncogenic development. Eph receptor A3 (EphA3) expression is associated with Esophageal cancer is the eighth most prevalent type of tumor promotion in certain types of cancer; however, it acts cancer worldwide (1,2), of which, esophageal squamous cell as a tumor suppressor in others. The expression levels of carcinoma (ESCC) is a predominant histological type (3). EphA3 and its effects on tumor progression in esophageal Despite advances in diagnostic tools, surgical techniques and squamous cell carcinoma (ESCC) cell lines were determined chemotherapy over the past few decades, the 5-year survival using reverse transcription-quantitative polymerase chain rate for patients with esophageal cancer ranges between reaction analysis and a Transwell invasion assay. The present 15 and 20% (4). Therefore, novel diagnostic tools, therapeutic study demonstrated that EphA3 expression was decreased strategies and molecular prognostic markers are urgently in ESCC tissues and cell lines. Treatment with the DNA required for this disease.
    [Show full text]
  • Detection of Pro Angiogenic and Inflammatory Biomarkers in Patients With
    www.nature.com/scientificreports OPEN Detection of pro angiogenic and infammatory biomarkers in patients with CKD Diana Jalal1,2,3*, Bridget Sanford4, Brandon Renner5, Patrick Ten Eyck6, Jennifer Laskowski5, James Cooper5, Mingyao Sun1, Yousef Zakharia7, Douglas Spitz7,9, Ayotunde Dokun8, Massimo Attanasio1, Kenneth Jones10 & Joshua M. Thurman5 Cardiovascular disease (CVD) is the most common cause of death in patients with native and post-transplant chronic kidney disease (CKD). To identify new biomarkers of vascular injury and infammation, we analyzed the proteome of plasma and circulating extracellular vesicles (EVs) in native and post-transplant CKD patients utilizing an aptamer-based assay. Proteins of angiogenesis were signifcantly higher in native and post-transplant CKD patients versus healthy controls. Ingenuity pathway analysis (IPA) indicated Ephrin receptor signaling, serine biosynthesis, and transforming growth factor-β as the top pathways activated in both CKD groups. Pro-infammatory proteins were signifcantly higher only in the EVs of native CKD patients. IPA indicated acute phase response signaling, insulin-like growth factor-1, tumor necrosis factor-α, and interleukin-6 pathway activation. These data indicate that pathways of angiogenesis and infammation are activated in CKD patients’ plasma and EVs, respectively. The pathways common in both native and post-transplant CKD may signal similar mechanisms of CVD. Approximately one in 10 individuals has chronic kidney disease (CKD) rendering CKD one of the most common diseases worldwide1. CKD is associated with a high burden of morbidity in the form of end stage kidney disease (ESKD) requiring dialysis or transplantation 2. Furthermore, patients with CKD are at signifcantly increased risk of death from cardiovascular disease (CVD)3,4.
    [Show full text]
  • Recombinant Human Epha8 Fc Chimera Catalog Number: 6828-A8
    Recombinant Human EphA8 Fc Chimera Catalog Number: 6828-A8 DESCRIPTION Source Mouse myeloma cell line, NS0­derived Human EphA8 Human IgG (Glu31­Thr542) IEGRMD 1 (Pro100­Lys330) Accession # NP_065387 N­terminus C­terminus N­terminal Sequence Glu31 Analysis Structure / Form Disulfide­linked homodimer Predicted Molecular 83.2 kDa (monomer) Mass SPECIFICATIONS SDS­PAGE 94 kDa, reducing conditions Activity Measured by its binding ability in a functional ELISA. When Recombinant Human EphA8 Fc Chimera is coated at 2 μg/mL (100 μL/well), the concentration of Biotinylayed Recombinant Human Ephrin­A5 Fc Chimera (Catalog # BT374) that produces 50% of the optimal binding response is 2­12 ng/mL. Endotoxin Level <0.01 EU per 1 μg of the protein by the LAL method. Purity >90%, by SDS­PAGE under reducing conditions and visualized by silver stain. Formulation Supplied as a 0.2 μm filtered solution in MES, NaCl, PEG, CHAPS and Imidazole. See Certificate of Analysis for details. PREPARATION AND STORAGE Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. Stability & Storage Use a manual defrost freezer and avoid repeated freeze­thaw cycles. l 12 months from date of receipt, ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after opening. l 3 months, ­20 to ­70 °C under sterile conditions after opening. BACKGROUND EphA8, also known as Hek3 and Eek, is a 120 kDa glycosylated member of the Eph family of transmembrane receptor tyrosine kinases (1, 2). The A and B classes of Eph proteins are distinguished by Ephrin ligand binding preference but have a common structural organization.
    [Show full text]