Functional and morphological plasticity of dendritic spines in the hippocampus [Elektronische Ressource] / vorgelegt von Nicola Eberhorn, geb. Tobisch

Nicola Eberhorn

Sujets

• Gesundheit

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2005
• Nombre de lectures: 17
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Functional and Morphological Plasticity of Dendritic
Spines in the Hippocampus

















Dissertation
zur Erlangung des Grades eines Doktors
der Naturwissenschaften

der Fakultät für Biologie
der Ludwig-Maximilians-Universität
München







vorgelegt von

Nicola Eberhorn, geb. Tobisch
aus München

2005






































1. Gutachter: Prof. Tobias Bonhoeffer
2. Gutachter: Prof. Benedikt Grothe

Tag der mündlichen Prüfung: 16.12.2005







To my family Acknowledgements

Grateful thanks to Prof. Tobias Bonhoeffer who gave me the opportunity to write my
PhD thesis in his laboratory and helped me at all times with his scientific and personal
support.
Thanks to my supervisor Dr. Sidney Cambridge who guided and supported me
throughout my time in the lab. I owe you a lot.
Thanks to Dr. Valentin Nägerl who shared his never ending enthusiasm for science with
me as a supervisor, a collaborator and a friend.
Thanks to Dr. Rüdiger Klein for being member of my thesis committee.
Thanks to Dr. Albrecht Kossel for proofreading this manuscript and for always being
there. You are the gute Seele.
Thanks to our excellent technicians Olusegun Momoh, Volker Staiger and Nicole Stöhr
for their help with the transgenic mice, the Gähwiler cultures and many more things.
Thanks to my colleagues: Dr. Albrecht Kossel, Dr. Harald Rösch, Dr. Marta Zagrebelsky
Holz, Dr. Christian Lohmann, Dr. Rosalina Fonseca, Susanne Lang for their support,
helpfulness, patience, humor and for inspiring discussions. Thanks to the Bonhoeffer lab.
Thanks to Max Sperling for his friendly help with data analysis and computer problems.
Thanks to Dr. Armin Seidl for his enduring friendship and support since my first day in
the lab.
Thanks to Bettina Reiter, Dr. Julia Schleypen and Cinthia Brise ňo who are the best
friends I could have.
Special thanks to my husband and friend Dr. Andreas Eberhorn. Together we reached a
lot.
Thanks and love to my son Luis Eberhorn.
Thanks to my mother Ursula Tobisch for her love and life-long support.
Table of contents
SUMMARY...............................................................................................................................................- 3 -
ZUSAMMENFASSUNG..........................................................................................................................- 5 -
INTRODUCTION....................................................................................................................................- 7 -
GENERAL INTRODUCTION....................................................................................................................... - 7 -
SYNAPSES............................................................................................................................................... - 7 -
SPINES .................................................................................................................................................... - 8 -
MORPHOLOGICAL PLASTICITY IN VIVO ................................................................................................. - 10 -
THE HIPPOCAMPUS ............................................................................................................................... - 10 -
HIPPOCAMPAL SLICE CULTURE ............................................................................................................. - 11 -
LONG TERM POTENTIATION (LTP)....................................................................................................... - 13 -
MORPHOLOGICAL PLASTICITY IN VITRO ............................................................................................... - 13 -
PROPOSAL OF THE SEQUENCE OF MORPHOLOGICAL EVENTS ................................................................. - 16 -
MORPHOLOGICAL CHANGES FOLLOWING LTP: THE OPEN QUESTIONS.................................................. - 17 -
PRUNING OF SPINES .............................................................................................................................. - 19 -
BIDIRECTIONAL MORPHOLOGICAL PLASTICITY..................................................................................... - 19 -
SUMMARY OF AIMS...........................................................................................................................- 21 -
CHAPTER 1: IMMUNOHISTOCHEMICAL CHARACTERIZATION OF ACTIVITY-
DEPENDENT SPINOGENESIS IN HIPPOCAMPAL NEURONS ..................................................- 22 -
SUMMARY ............................................................................................................................................ - 23 -
INTRODUCTION..................................................................................................................................... - 25 -
RESULTS............................................................................................................................................... - 28 -
Time-lapse imaging.........................................................................................................................- 28 -
Posthoc immunohistochemistry.......................................................................................................- 29 -
Three-dimensional immunohistochemistry in organotypic slice cultures .......................................- 29 -
Newly born spines often contact presynaptic terminals..................................................................- 31 -
Quantitative and qualitative measurement of the time-course of spine- α-synapsin colocalization - 32 -
Reidentified newly grown spines mostly lack postsynaptic specializations ....................................- 33 -
Quantitative and qualitative measurement of the time-course of spine-GluR2 colocalization .......- 34 -
DISCUSSION.......................................................................................................................................... - 35 -
METHODS............................................................................................................................................. - 42 -
Organotypic hippocampal cultures.................................................................................................- 42 -
Recording solutions. .......................................................................................................................- 42 -
Electrophysiology. ..........................................................................................................................- 43 -
Microscopy......................................................................................................................................- 43 -
Image analysis I. .............................................................................................................................- 44 -
Posthoc-immunohistochemistry and laser-scanning confocal microscopy.....................................- 45 -
Image analysis II.............................................................................................................................- 45 -
FIGURES ............................................................................................................................................... - 47 -
FIGURE LEGENDS ................................................................................................................................. - 55 -
Figure 1: Time-lapse imaging of dendritic fine structure and their reidentification after fixation and
immunohistochemistry. ...................................................................................................................- 55 -
Figure 2: Quantification of colocalization of mature spines and immunohistochemical staining..- 55 -
Figure 3: Reidentified newly grown spines often contact presynaptic terminals............................- 56 -
Figure 4: Histogram and time-course of the colocalization between activity induced newly grown
spines and immunopositive puncta .................................................................................................- 56 -
Figure 5: Reidentified newly grown spines mostly lack postsynaptic specializations ....................- 56 -
Figure 6: Model for synaptogenesis on de novo grown spines.......................................................- 57 -
Supplemental Figure 1: Examples of activity induced newly grown spines and their extend of
colocalization with anti-synapsin stainings ....................................................................................- 57 -
Supplemental Figure 2: Histogram and time-course of the colocalization between de novo grown
spines and immunopositive puncta; combined results of different experimental conditions. .........- 57 -
- 1 -REFERENCES...........................................................................................................