Differentiation of dendrites and the analysis of spine-like structures on lobula plate tangential cells in Drosophila melanogaster [Elektronische Ressource] / vorgelegt von Ewa Joanna Koper

Ewa Koper

Sujets

• Biologie

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2007
• Nombre de lectures: 24
• Langue: Deutsch
• Poids de l'ouvrage: 4 Mo

Extrait

Differentiation of Dendrites
and the Analysis of Spine- Like Structures
on Lobula Plate Tangential Cells
in Drosophila melanogaster





Dissertation

Zur Erlangung des Doktorgrades
der Naturwissenschaften (Dr. rer. nat)
der Fakultät für Biologie
der Ludwig- Maximilians- Universität München

Angefertigt am Max Planck Institut für Neurobiologie,
Abteilung Molekulare Neurobiologie,
Abteilungsgruppe Dendritische Differenzierung

Vorgelegt von
Ewa Joanna Koper
München 2007



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Hiermit, erkläre ich, dass ich die vorliegende Dissertation selbständig und ohne unerlaubte
Hilfe angefertigt habe. Sämtliche Experimente wurden von mir selbst durchgeführt, au βer
wenn explizit auf Dritte verwiesen wird. Ich habe weder anderweitig versucht, eine
Dissertation oder Teile einer Dissertation einzureichen bzw. einer Prüfungskommission
vorzulegen, noch eine Doktorprüfung durchzuführen.

München, den.............................. ...................................................
(Unterschrift)














1st Gutachter: Prof. Benedikt Grothe
2nd Gutachter: Prof. Ruediger Klein
Tag der mündlichen Prüfung: 08.05.2007









Die vorliegende Arbeit wurde zwischen Juni 2003 und Dezember 2006 unter der Leitung
von Dr. Gaia Tavosanis am Max- Planck Institut für Neurobiologie in Martinstried
durchgeführt.








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Table of Contents


Table of Contents


Table of Contents ..................................................................................................i

Figures and Tables................................................................................................................... iv
Abbreviations............................................................................................................................ vi

Summary ...............................................................................................................1

1 Introduction ....................................................................................................3

Dendrite Morphology.............................................................................................................. 3
Factors involved in Dendritogenesis...................................................................................... 3
Building the Dendritic Tree.5
Dendritic Cytoskeleton ............................................................................................................ 7
Dendritic Spines........................................................................................................................ 10
Dendritic Filopodia .................................................................................................................. 15
Synaptic Contacts..................................................................................................................... 16
Anatomy of Lobula Plate Tangential Cells (LPTCs)............................................................ 18
Visual System of a Fly.... 20
LPTCs as a Part of Neuronal Circuitry ................................................................................. 22
Development of the Fly Central Nervous System............................................................... 23

2 Materials and Methods...................................................................................25

Fly Stocks................................................................................................................................... 25
Antibodies............. 26
Instruments ................................................................................................................................. 27
Consumables......... 27
iTable of Contents


Solution and Media ................................................................................................................. 28
Fly Maintenance ....................................................................................................................... 28
Staging of Animals...... 29
Dissection of Adult Fly Brains................................................................................................. 29 n of Larval and Pupal Fly Brains............................................................................ 29
Sample Preparation... 30
Gal4- UAS System................................................................................................................... 30
Confocal Image Acquisition.................................................................................................... 34
MARCM Technique................................................................................................................... 34
Analysis of the Images ............................................................................................................ 35
Quantitative Morphological Analysis of Spine-Like Structures........................................ 36
Immunohistochemical Methods................................................................................................ 37
Electron Microscopy................................................................................................................. 38
Culturing Fly Brains .................................................................................................................. 38

3 Results..........................................................................................................40

Developmental Time Points..................................................................................................... 40
Late Larval Stage .................................................................................................................... 42
Early Pupal Stage...... 42
Intermediate Pupal Stages..................................................................................................... 44
Late Pupal Stages...... 46
Refinement Phase..................................................................................................................... 48
Stability Phase.......................................................................................................................... 48
Development of LPTC Dendrites- Summary ........................................................................ 49
Subcellular Localization of Actin ........................................................................................... 50
LPTC Dendrites Bear Spine- like Protrusions....................................................................... 52
LPTC Spine- like Structures are Enriched in Actin............................................................... 53
Density and Length of LPTC Spine- like Structures............................................................. 56
Morphological Categories of LPTC Spine- like Structures................................................ 57
Synaptic Input onto LPTC Spine- like Structures ................................................................. 59
iiTable of Contents
Ultrastructure of LPTC Spine- like Structures....................................................................... 61
Genetic Modifications of LPTC Spine- like Structures ....................................................... 63
Spine- Like Structures on other Drosophila Neurons .......................................................... 67
Motility of LPTC Spine- like Structures ................................................................................. 68

4 Discussion ......................................................................................................70

Why study Dendritogenesis in LPTCs?.................................................................................. 71
LPTC Spine- like Structures are Actin Enriched ................................................................... 76
Why study Spines in Drosophila? .......................................................................................... 78
Learning in Drosophila............................................................................................................. 79
Are LPTC Spine- like Structures Plastic?............................................................................... 80
Are the synaptic contacts on LPTC Spine- like Structures Active?.................................... 82
Genetical Manipulations of LPTC Spine- like Structures................................................... 84
Differences between LPTC Spine- like Structures and Vertebrate Spines .........