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Supervised by Docent Juha Partanen Institute of Biotechnology University of Helsinki Finland Reviewed by Professor Eero Castrén Neuroscience Center University of Helsinki Finland Professor Seppo Vainio Oulu Biocenter University of Oulu Finland Opponent Ph.D. Mark Lewandoski National Cancer Institute Frederick Cancer Research & Development Center U.S. Custos Professor Jim Schröder Department of Biological and Environmental Sciences University of Helsinki Finland ISSN 1795-7079 ISBN 978-952-10-4034-4 (paperback) ISBN 978-952-10-4035-1 (PDF, http://ethesis.helsinki.fi ) Edita, Helsinki 2007 If we knew what we were doing we wouldn’t call it science. Albert Einstein TABLE OF CONTENTS ABBREVIATIONS List of original publications I. Jukkola T*, Lahti L* Naserke T, Wurst W, Partanen J. FGF regulated gene- expression and neuronal differentiation in the developing midbrain-hindbrain region. Developmental Biology. 2006 Sep 1; 297(1): 141-57. II. Jukkola T*, Sinjushina N*, Partanen J. Drapc1 expression during mouse embryonic development. Gene Expression Patterns. 2004 Oct; 4(6): 755-62. III. Trokovic R*, Jukkola T*, Saarimäki J, Peltopuro P, Naserke T, Weisenhorn DM, Trokovic N, Wurst W, Partanen J. Fgfr1-dependent boundary cells between developing mid- and hindbrain. Developmental Biology. 2005 Feb 15; 278(2): 428- 39. IV. Kala K, Jukkola T, Pata I, Partanen J. Analysis of the midbrain-hindbrain boundary cell fate using a boundary cell-specifi c Cre-mouse line. Manuscript submitted to Genesis. SUMMARY REVIEW OF THE LITERATURE ............................................................................. 1 1. The early brain development ..................................................................................... 1 1.1. Neural induction and neurulation .......................................................................1 1.2. Molecular events of neural induction and neurulation ....................................... 1 2. Mechanisms of brain patterning ................................................................................. 2 2.1. Anterioposterior patterning ................................................................................ 2 2.2. Isthmic organizer ................................................................................................ 2 2.2.1. Induction and localization of isthmic organizer ....................................... 5 2.3. Neuronal development and regionalization of the midbrain .............................. 9 2.4. Neuronal development and regionalization of the hindbrain ........................... 11 2.4.1. Segmentation and compartmentalization in hindbrain ........................... 12 3. Intercellular communication in the midbrain-R1 region .......................................... 14 3.1. FGF signaling in the midbrain-R1 development .............................................. 14 3.1.1. The FGF and FGFR molecules ............................................................... 14 3.1.2. Expression patterns of FGFs and FGFRs in the mouse midbrain-R1 .......................................................................................... 15 3.1.3. FGFRs in the mouse midbrain-R1 development .................................... 16 3.1.4. FGFR signal transduction ....................................................................... 17 3.2. WNT signaling in the midbrain-R1 development ............................................. 21 4. Cell proliferation in the developing brain .................................................................23 4.1. Interkinetic nuclear migration .......................................................................... 23 4.2. Cell cycle regulators ......................................................................................... 25 4.2.1. Cyclins, Cdks and their inhibitors .......................................................... 25 4.2.2. Cyclins and Cdks in neuronal development ........................................... 26 4.2.3. Cdk inhibitors in neuronal development ................................................ 27 4.3. Mechanisms controlling the cell polarity and asymmetric cleavage ................ 28 4.4. Cellular proliferation and regulation in the rhombomeric boundaries ............ 28 AIMS OF THE STUDY .............................................................................................. 30 MATERIALS AND METHODS ................................................................................ 31 RESULTS AND DISCUSSION .................................................................................. 34 1. Microarray analysis of Fgfr1 CKO mutants (I-III) .................................................. 34 2. Identifi cation of FGF regulated genes at the midbrain-R1 region (I, III and unpublished) ............................................................................................ 43 2.1. Down-regulated genes (I and unpublished) ..................................................... 43 2.1.1. Down-regulation of FGF-FGFR signaling components at E10.5 (I, III) ................................................................................................................ 43 2.1.2. Cnpy1 as a novel regulator of FGF signaling at the midbrain-R1 region (I and unpublished) ............................................................................... 45 2.1.3. Down-regulated genes expressed as a narrow stripe near the MHB (I and unpublished) .......................................................................................... 46 2.1.4. Other down-regulated genes in the Fgfr1 CKO mutants (I) ................... 48 2.2. Up-regulated genes show loss of expression gradients in Fgfr1 CKO mutant embryos (I).............................................................................................48 3. Neuronal progenitor cell populations are affected in Fgfr1 CKO mutant embryos (I) ..............................................................................................................