Holdings: Modeling human zinc biology using iPSC derived cells

Modeling human zinc biology using iPSC derived cells:

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Main Author:	Pfänder, Stefanie (Author)
Format:	Thesis Book
Language:	English
Published:	Ulm 2016
Subjects:	Hochschulschrift
Online Access:	Inhaltsverzeichnis Inhaltsverzeichnis
Physical Description:	VII, 134 Blätter Illustrationen 30 cm

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adam_text	ULM UNIVERSITY INSTITUTE FOR ANATOMY AND CELL BIOLOGY DIRECTOR: PROF. DR. MED. TOBIAS M. BOECKERS MODELING HUMAN ZINC BIOLOGY USING IPSC DERIVED CELLS DISSERTATION SUBMITTED IN PARTIAL FULFILLM ENT OF THE REQUIREMENTS FOR THE DEGREE OF,,DOCTOR RERUM NATURALIUM (DR. RER. NAT.) OF THE INTERNATIONAL GRADUATE SCHOOL IN MOLECULAR MEDICINE ULM STEFANIE PFAENDER FROM BIBERACH AN DER RISS 2016 CONTENTS ABBREVIATIONS........................................................................................................................................... ILL 1 INTRODUCTION..................................................................................................................................... 1 1.1 AUTISM SPECTRUM DISORDERS......................................................................................................... 1 1.1.1 CAUSES.................................................................................................................................... 2 1.1.2 PHELAN-MCDERMID SYNDROME...........................................................................................10 1.1.3 INDUCED PLURIPOTENT STEM CELLS ........................................... 15 1.2 AIMS OF THIS THESIS...................................................................................................................... 18 2 MATERIAL & METHODS...................................................................................................... I.............20 2.1 M ATERIAL....................................................................................................................................... 20 2.1.1 CHEMICALS........................................................................................................................... 20 2.1.2 CELL CULTURE REAGENTS AND GROWTH FACTORS..................................................................... 21 2.1.3 KITS....................................................................................................................................... 23 2.1.4 MARKERS.............................................................................................................................. 24 2.1.5 CONSUMABLES......................................................................................................................24 2.1.6 EQUIPMENT.......................................................................................................................... 25 2.1.7 SOFTWARE............................................................................................................................. 26 2.1.8 PRIMER..................................................................................................................................26 2.1.9 ANTIBODIES.......................................................................................................................... 28 2.1.10 CELL CULTURE M EDIA............................................................................................................. 30 2.1.11 BUFFERS AND SOLUTIONS.........................................................................................................31 2.2 METHODS...................................................................................................................................... 32 2.2.1 PATIENT STUDIES................................................................................................................... 32 2.2.2 CELL CULTURE......................................................................................................................... 33 2.2.3 ANALYSIS............................................................................................................................... 37 2.2.4 STATISTICAL ANALYSIS..............................................................................................................42 3 RESULTS............................................................................................................................................... 43 3.1 ZINC DEFICIENCY IN PHELAN-MCDERMID SYNDROME.....................................................................43 3.1.1 ZINC DEFICIENCY IN PHELAN-MCDERMID SYNDROME AS ASSESSED BY ATOMIC ABSORPTION SPECTROSCOPY OF BLOOD SAMPLES IS ASSOCIATED WITH CERTAIN SYMPTOMS ...................................... 43 3.1.2 ZINC DEFICIENCY AND ABNORMALLY LOW ZN/CU RATIOS IN PHELAN-MCDERMID SYNDROME CONFIRMED BY INDUCTIVELY-COUPLED PLASMA MASS SPECTROMETRY OF HAIR SAMPLES ........................ 46 3.2 CAUSES OF ZINC DEFICIENCY IN PHELAN-MCDERMID SYNDROME..................................................... 50 3.2.1 ALTERED ZINC HOMEOSTASIS PROTEIN EXPRESSION IN IPSC DERIVED ENTEROCYTES FROM PMDS PATIENTS..................................................................................................................................... 50 3.2.2 ALTERED REGULATION OF ZINC HOMEOSTASIS GENE EXPRESSION.............................................55 3.2.3 MEMBRANE-ASSOCIATED COLOCALIZATION OF ZIPS WITH SHANK3 IS ALTERED IN PMDS ...... 57 3.2.4 INTERACTION OF ZIPS WITH SHANK3..................................................................................... 59 3.3 CONSEQUENCES OF ZINC DEFICIENCY IN NEURODEVELOPMENT ..................................................... 62 3.3.1 REGULATION OF ZINC LEVELS IN NEURODEVELOPMENT ............................................................ 62 3.3.2 ZINC DEFICIENCY CHANGES EXPRESSION LEVELS OF ZINC HOMEOSTASIS GENES IN NEURODEVELOPMENT..............................................................................................................................67 3.3.3 ZINC DEFICIENCY IMPAIRS VIABILITY IN MOTOR NEURONAL DIFFERENTIATION ........................... 69 3.3.4 ZINC DEFICIENCY IN MOTOR NEURONAL DEVELOPMENT DOES NOT ALTER MORPHOLOGY OF MATURE MOTOR NEURONS.......................................................................................................................71 3.3.5 ZINC DEFICIENCY IN MOTOR NEURONAL DEVELOPMENT AFFECTS ACETYLCHOLINE AND GLUTAMATE RECEPTOR SIGNALING IN MATURE MOTOR NEURONS..................................................................................72 4 DISCUSSION......................................................................................................................................... 78 4.1 ZINC DEFICIENCY AND LOW ZN/CU RATIOS ARE ASSOCIATED WITH PMDS AND CERTAIN CO MORBIDITIES IN PMDS................................................................................................................................78 4.2 CAUSES FOR ZINC DEFICIENCY IN PMDS ......................................................................................... 82 4.2.1 ALTERED EXPRESSION OF ZINC HOMEOSTASIS PROTEINS IN ENTEROCYTES OF PMDS................83 4.2.2 ALTERED LOCALIZATION OF ZINC HOMEOSTASIS PROTEINS IN ENTEROCYTES OF PMDS .............. 84 4.2.3 ALTERATIONS IN ZIP2 AND ZIP4 ARE SHANK3 DEPENDENT .................................................. 85 4.2.4 INTERACTION OF ZIP WITH SHANK3 IN ENTEROCYTES............................................................. 86 4.3 REGULATION OF ZINC HOMEOSTASIS PROTEIN EXPRESSION IN MOTOR NEURON DEVELOPMENT ....... 87 4.4 ZINC DEFICIENCY INDUCES CELL DEATH IN MOTOR NEURONAL DEVELOPMENT .................................. 89 4.5 ZINC DEPLETION INDUCES ALTERATIONS IN SYNAPSE RECEPTOR COMPOSITION AND FUNCTION ......... 90 4.6 LIMITATIONS...................................................................................................................................95 4.7 CONCLUSION...................................................................................................................................96 5 SUMMARY........................................................................................................................................... 98 6 REFERENCES...................................................................................................................................... 100 7 APPENDIX.......................................................................................................................................... 124 8 PEER-REVIEWED PUBLICATIONS.................................................................................................... 132 9 AKNOWLEDGEMENTS......................................................................................................................133 10 STATUTORY DECLARATION.........................................................................................................134
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spelling	Pfänder, Stefanie Verfasser aut Modeling human zinc biology using iPSC derived cells Stefanie Pfänder Ulm 2016 VII, 134 Blätter Illustrationen 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation International Graduate School in Molecular Medicine Ulm 2016 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1132590280/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029844503&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Pfänder, Stefanie Modeling human zinc biology using iPSC derived cells
subject_GND	(DE-588)4113937-9
title	Modeling human zinc biology using iPSC derived cells
title_auth	Modeling human zinc biology using iPSC derived cells
title_exact_search	Modeling human zinc biology using iPSC derived cells
title_full	Modeling human zinc biology using iPSC derived cells Stefanie Pfänder
title_fullStr	Modeling human zinc biology using iPSC derived cells Stefanie Pfänder
title_full_unstemmed	Modeling human zinc biology using iPSC derived cells Stefanie Pfänder
title_short	Modeling human zinc biology using iPSC derived cells
title_sort	modeling human zinc biology using ipsc derived cells
topic_facet	Hochschulschrift
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