Transcriptional regulation of axonal pathfinding [Elektronische Ressource] : Foxb1 and the mammillothalamic tract / von Qiuhong Jiang

Qjiang

Sujets

• Biologie

Informations

• Publié par: gottfried_wilhelm_leibniz_universitat_hannover
• Publié le: 01 janvier 2004
• Nombre de lectures: 20
• Langue: English
• Poids de l'ouvrage: 9 Mo

Extrait

TRANSCRIPTIONAL REGULATION OF
AXONAL PATHFINDING: FOXB1 AND THE
MAMMILLOTHALAMIC TRACT





Vom Fachbereich Chemie der Universität Hannover
zur Erlangung des Grades

Doktor der Naturwissenschaften
Dr. rer. nat.

genehmigte Dissertation
von

M.Sc. Biochemistry Qiuhong Jiang
geboren am 8.Dezember 1972 in Hunan Province, China



2004

























Referent:
Prof. Dr. Walter Müller
Medizinische Hochschule Hannover

Korreferent:
Prof. Dr. Gregor Eichele
Max-Planck-Institut für experimentelle Endokrinologie, Hannover

Tag der Promotion: 15. November 2004



ACKNOWLEDGEMENTS

Before I start, I would like to acknowledge all those who have helped and supported me while I
was working on this PhD:

First of all, I would like to thank Prof. Dr. Gregor Eichele for the opportunity to work in this
Department, and for making available to me everything that was needed for my research. Above
all, his personal interest in the process of the work, fruitful discussions and guidance for the
independent scientific work contributed crucially to the successful execution of my project.

I am very grateful to Prof. Dr. Walter Müller for friendly accepting to assume the task of Official
Supervisor of my work in representation of the Chemistry Department of the University of
Hannover, and for his support in dealing with University bureaucracy.

I particularly appreciate Dr. Gonzalo Alvarez-Bolado for his excellent scientific supervision, and
for offering me the opportunity to work in his group, as well as for proposing me such an
intellectually stimulating subject for my PhD. His encouragement and guidance have been
essential for me to take this difficult and fascinating project to good end. I will always remember
his seminars on developmental neurobiology, which have been so important for me to enter into
this exciting field.

The help, advice and friendship of Dr. Axel Visel have been very important to me in everyday
life in the laboratory, in setting up difficult experiments, proofreading my thesis and in
overcoming the bureaucratic difficulties as a foreigner student.

I would like to thank the administration, especially Carsten Gottschalk for patiently helping with
formalities.

All through these years in Hannover I have received intelligent advice and kind assistance from
Dr. Pablo Szendro, Dr. Xunlei Zhou, Dr. Miki Tsukada. I am indebted to them.


I thank Barbara Fischer, Uwe Grunenberg, Ana Martínez-Hernández, Kornelia Maslo, Polina
Spies and Christine Zwingmann for their help with in situ hybridization work.

My friends and colleagues, the PhD students and postdocs of the Department, Dr. Henrik Oster,
Diya Abraham, Carsten Möller (Dr. Möller), Murat Yaylaoglu (Dr. Yaylaoglu), Nora Szabó,
Aravind Sekhar, Vladimira Jakubcakova, Frank Sacher, Judit Oldekamp (Dr. Oldekamp), Lars
Geffers, Marei Warnecke (Dr. Warnecke), Judit Kovac, Christopher Esk, Stephan Busche, Murat
Cankaya, Marían Comas, Jens Mittag, Katja Hübel, Johanna Rose, Andrew Titmus, and Tianyu
Zhao, contributed their help to the pleasant working climate which I could enjoy at our Institute.

The present work would have been impossible without the support of the Mouse House
personnel, Ina Klebba, Alexandra Meneking, Nadine Naujokat and Hans Peter Bader, working
under the wise direction of Dr. Michael Leitges.

Claus Ebert, Sarah Herzog and Markus Uhr solved promptly and expertly all my frequent
computer problems, and for this they deserve many thanks.

I thank Dr. T. Ciossek and Dr. P. Vanderhaeghen (Belgium) for kindly sharing the EphA7 mutant
mouse line with us.

Finally, I am grateful to my husband, my parents, and my country, China, for all what they have
done for me.

TABLE OF CONTENTS
TABLE OF CONTENTS
ZUSAMMENFASSUNG......................................................................................................................10
ABSTRACT ..........................................................................................................................................11
1 INTRODUCTION........................................................................................................................12
1.1 REGULATION OF GENE EXPRESSION DURING DEVELOPMENT .................................. 12
1.1.1 Regulatory transcription factors............................................................................................. 13
1.1.2 Transcription factor families .................................................................................................. 13
1.2 FORKHEAD FAMILY OF TRANSCRIPTION FACTORS ...................................................... 13
1.2.1 General aspects of forkhead proteins ..................................................................................... 13
1.2.1.1 Nomenclature................................................................................................................................ 14
1.2.1.2 Structure of the forkhead domain (DNA-binding domain)............................................................ 17
1.2.1.3 DNA binding specificity ............................................................................................................... 18
1.2.1.4 Transcription effector domains ..................................................................................................... 19
1.2.1.5 Forkhead genes in development.................................................................................................... 19
1.2.2 Characteristics of forkhead transcription factor Foxb1 ......................................................... 20
1.3 HYPOTHALAMUS ................................................................................................................... 22
1.3.1 General aspects about hypothalamus..................................................................................... 22
1.3.2 Mammillary body.................................................................................................................... 22
1.3.3 Foxb1 function in the hypothalamus....................................................................................... 24
1.4 AXONAL NAVIGATION.......................................................................................................... 26
1.4.1 Cellular interactions that guide axons.................................................................................... 26
1.4.2 Ephrins and Eph ..................................................................................................................... 27
1.5 MICROARRAY TECHNOLOGY.............................................................................................. 31
1.5.1 Genomics and DNA arrays..................................................................................................... 31
1.5.2 Affymetrix GeneChips............................................................................................................. 31
1.5.3 Application of Affymetrix GeneChips to brain development questions, and...............................
attending technical challenges................................................................................................ 33
1.5.4 Validation of microarray data ................................................................................................ 34
2 MATERIALS AND METHODS ................................................................................................36
2.1 ROUTINE FOR ANIMAL WORK....................................................................................................... 36
2.2 GENERAL MOLECULAR BIOLOGICAL METHODS ............................................................................ 36
2.2.1 Isolation of nucleic acids........................................................................................................ 37
5 TABLE OF CONTENTS
2.2.1.1 Isolation of genomic DNA from tissue samples............................................................................ 37
2.2.1.2 Small-scale isolation of plasmid DNA.......................................................................................... 37
2.2.1.3 Midi preparation of plasmid DNA ................................................................................................ 37
2.2.1.4 EndoFree plasmid Maxi protocol.................................................................................................. 38
2.2.1.5 Isolation of DNA fragments from agarose gel .............................................................................. 39
2.2.1.6 Direct purification of PCR products.............................................................................................. 39
2.2.1.7 Isolation of total RNA from cells.................................................................................................. 40
2.2.1.8 Isolation of poly(A)-enriched RNA .............................................................................................. 40
2.2.1.9 R