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Die Reorganisation der Fascia dentata der Ratte nach entorhinaler Denervierung [Elektronische Ressource] : eine regulatorische Rolle für das Chondroitinsulfat Proteoglykan NG2 / von Doris Dehn

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93 pages
Die Reorganisation der Fascia dentata der Ratte nach entorhinaler Denervierung: eine regulatorische Rolle für das Chondroitinsulfat Proteoglykan NG2 Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften vorgelegt beim Fachbereich Biowissenschaften der Johann Wolfgang Goethe-Universität in Frankfurt am Main von Doris Dehn aus Hattingen Frankfurt 2005 (D30) Vom Fachbereich Biowissenschaften der Johann Wolfgang Goethe-Universität als Dissertation angenommen. Dekan: Professor Dr. Rüdiger Wittig Gutachter: Professor Dr. Herbert Zimmermann Professor Dr. Thomas Deller Datum der Disputation: The reorganization of the rat dentate gyrus following entorhinal denervation: a regulatory role for the chondroitin sulfate proteoglycan NG2 Dissertation submitted to the Faculty of Biological Sciences of the Johann Wolfgang Goethe-University of Frankfurt/Main, Germany for the degree of Doctor of Natural Sciences (doctor philosphiae naturalis, Dr. phil.nat.) presented by Doris Dehn (Hattingen/Ruhr) Frankfurt/Main, 2005 Accepted by the Faculty of Biological Sciences of the Johann Wolfgang Goethe-University Frankfurt/Main, Germany Dean: Professor Dr. Rüdiger Wittig Referees: Professor Dr. Herbert Zimmermann Professor Dr.
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Die Reorganisation der Fascia dentata der Ratte
nach entorhinaler Denervierung:
eine regulatorische Rolle für das Chondroitinsulfat Proteoglykan NG2
Dissertationzur Erlangung des Doktorgrades der Naturwissenschaften
vorgelegt beim Fachbereich Biowissenschaften der Johann Wolfgang Goethe-Universität in Frankfurt am Main
von Doris Dehn aus Hattingen
Frankfurt 2005 (D30)
Vom Fachbereich Biowissenschaften der
Johann Wolfgang Goethe-Universität als Dissertation angenommen.
Dekan:
Gutachter:
Datum der Disputation:
Professor Dr. Rüdiger Wittig
Professor Dr. Herbert Zimmermann
Professor Dr. Thomas Deller
The reorganization of the rat dentate gyrus
following entorhinal denervation:
a regulatory role for the chondroitin sulfate proteoglycan NG2
Dissertation
submitted to the Faculty of Biological Sciences
of the Johann Wolfgang Goethe-University of Frankfurt/Main, Germany
for the degree of
Doctor of Natural Sciences
(doctor philosphiae naturalis, Dr. phil.nat.)
presented by
Doris Dehn
(Hattingen/Ruhr)
Frankfurt/Main, 2005
Accepted by the Faculty of Biological Sciences
of the Johann Wolfgang Goethe-University
Frankfurt/Main, Germany
Dean:
Referees:
Oral examination:
Professor Dr. Rüdiger Wittig
Professor Dr. Herbert Zimmermann
Professor Dr. Thomas Deller
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'What is this,' said the Leopard, 'that is so 'sclusively dark,
and yet so full of little pieces of light?'
Rudyard Kipling, How the leopard got his spots
ABSTRACT
INTRODUCTION
REORGANISATION PROCESSES AFTER CENTRAL NERVOUS SYSTEM INJURYAXONAL SPROUTING AFTER ENTORHINAL CORTEX LESIONMOLECULAR REGULATION OF SPROUTINGCHONDROITINSULFATE PROTEOGLYCANS(CSPGS) AMEMBER OF THECSPGFAMILY: NG2 AIMS OF THE STUDY
MATERIALS AND METHODS
ANIMALS,SURGICAL PROCEDURES,AND TISSUE PREPARATIONANIMALSENTORHINAL CORTEX LESIONTISSUE PREPARATION
CONTROL OF LESION QUALITYFLUORO-JADEBANDACHESTAINING ON FRONTAL SECTIONS OF RAT DENTATE GYRUSCOMBINED LASERMICRODISSECTION(LMD)AND QUANTITATIVERT-PCR (QPCR) FLOW CHART OF THE COMBINEDLMDAND QPCRPROTOCOLPROTOCOL FORRNAISOLATION USING THELMDANDRNEASY®MICROKIT (QUIAGEN)/TAQMANREVERSETRANSCRIPTIONREAGENTS(APPLIEDBIOSYSTEMS) QUANTITATIVE REVERSE TRANSCRIPTION-PCR (QPCR) IMMUNOHISTOCHEMISTRYSTAINING PROTOCOLSBUFFER SOLUTIONS
TRANSMISSION ELECTRON MICROSCOPYBROMO-DEOXYURIDINE INJECTIONDIGITAL PROCESSING OF ILLUSTRATIONS
RESULTS
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2222 22 23
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29 31 3233 35 363636
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AMOUNT OFNG2MRNAIS SIGNIFICANTLY INCREASED IN THE DENERVATED OUTER MOLECULAR LAYER 37NG2-POSITIVE CELLS ARE REGULARLY DISTRIBUTED IN THE DENTATE GYRUS OF CONTROL RATS 39STAINING OFNG2-POSITIVE CELLS IS ENHANCED IN THE DENERVATED OUTER MO CULA 39LE R LAYER TOF THE DENERVATED OUTER MOLECULAR LAYER IS ENRICHED WITHHE NEUROPIL NG2 FOLLOWING ENTORHINAL CORTEX LESION 42ELECTRON MICROSCOPY OFNG2-POSITIVE CELLS IN THE OUTER MOLECULAR LAYER: IMMUNOPRECIPITATE IN THE EXTRACELLULAR MATRIX SURROUNDING GLIAL AND NEURONAL PROFILES 44NG2-POSITIVE CELLS DO NOT EXPRESS ASTROGLIAL,MICROGLIAL,AND ADULT OLIGODENDROGLIAL MARKERS 47ASUBPOPULATION OFNG2-POSITIVE CELLS IS GENERATED POSTLESION IN THE OUTER OLECULAR LAY 50M ER
DISCUSSION
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SUMMARY OF RESULTS 52METHODOLOGICAL CONSIDERATIONS 52COMBINED LASERMICRODISSECTION AND QUANTITATIVERT-PCR 53 IMMUNOHISTOCHEMISTRY 55 DOESNG2AFFECT AXONS WITHIN THE ZONE OF DENERVATION? REGULATION OF BRANCHING AND AXONAL COMPLEXITY 56DOESNG2AFFECT AXONS OUTSIDE THE ZONE OF SPROUTING? REGULATION OF A LAYER-SPECIFIC SPROUTING RESPONSE 58SECRETION OF CLEAVEDNG2INTO THE ZONE OF SPROUTING 59REGULATION OFECLINDUCED AXONAL GROWTH:THE ROLE OF THE EXTRACELLULAR MATRIX 60NG2-POSITIVE CELLS:A FOURTH GLIAL CELL TYPE IN THE BRAIN OF ADULT RATS? 60
SUMMARY AND CONCLUSION
64
REFERENCE LIST
CURRICULUM VITAE
PUBLICATIONS
ACKNOWLEDGEMENT
ZUSAMMENFASSUNG (DEUTSCH)
LEBENSLAUF
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