Migration of branchiomotor neurons to the alar plate in the chicken embryo [Elektronische Ressource] / vorgelegt von Min Jeong Ju

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Informations

• Publié par: universitat_duisburg-essen
• Publié le: 01 janvier 2004
• Nombre de lectures: 18
• Langue: English
• Poids de l'ouvrage: 3 Mo

Extrait

Medizinische Fakultät
der
Universität Duisburg-Essen


Aus dem Institut für Anatomie




Migration of Branchiomotor Neurons to the Alar Plate

in the Chicken Embryo






Inaugural-Dissertation
zur
Erlangung des Doktorgrades der Medizin
durch die Medizinische Fakultät
der Universität Duisburg-Essen





Vorgelegt von
Min Jeong Ju
aus Seoul
2003


1


























Dekan: Univ.-Prof. Dr. H. Grosse-Wilde
1. Gutachter: Univ. Esche
2. Gutachter: Univ.-Prof. Dr. Dr. C. Redies
Tag der mündlichen Prüfung: 28, Mai, 2004
2
Teile dieser Arbeit wurden veröffentlicht in:

Originalveröffentlichung:
Ju, M. J., Aroca, P., Puelles, L., Redies, C.
Molecular profiling indicates avian branchiomotor nuclei invade hindbrain
alar plate
Zur Veröffentlichung eingereicht

Kurzveröffentlichungen:
Posterveröffentlichung auf dem 1. Forschungstag der medizinischen
Fakultät in Essen (13. Juni 2002):
Ju, M. J., Puelles, L., Redies, C. (2002)
Differential expression of two cadherins in the hindbrain and spinal cord of
the chicken embryo

Posterveröffentlichung auf dem 2. Forschungstag der medizinischen
Fakultät in Essen (18. Juli 2003):
Ju, M. J., Aroca, P., Puelles, L., Redies, C. (2003)
Molecular profiling indicates avian branchiomotor nuclei invade hindbrain
alar plate

Vortrag auf der 20. Arbeitstagung der Anatomischen Gesellschaft in
Würzburg (1.-3. Oktober 2003):
Ju, M. J., Aroca, P., Puelles, L., Redies, C. (2003)
3
Molecular profiling indicates that branchiomotor nuclei come to reside in
the alar plate of the avian hindbrain
4
Table of Contents Page
List ofAbreviatons 8ist ofIlustraions 1

1  Introduction 13

1.1 Development of spinal cord and hindbrain from 13
neural tube
1.2 Dorsoventral patterning in the spinal cord and hindbrain 17
1.3 Markers for dorsoventral patterning and development 18
of branchiomotor neurons
1.4 Cadherins 20
1.4.1 Structure of cadherins 20
1.4.2 Roles of cadherins in development 23
1.4.3 Cadherins and disease 27

2 Material and Methods 31

2.1 Chemicals and instruments 31
2.1 Chemicals 31
2.12 Instruments 3
2.1.3 Computer software and hardware 33

2.2 Solutions for immunohistochemistry 34
5
2.2.1 HEPES-buffered stock solution (10X) 34
2+ 2+ 2.2.2 HEPES-buffered solution with Ca and Mg 34
2.2.3 TBS stock solution (10X) 35
2.2.4 TBS solution (1X) 35
2.5 Sucrose olution 5
2.2.6 PBS stock solution (10X) 35
2.2.7 PBT solution 36
2.2.8 4% PFA in HEPES-buffered solution 36
2.9 ABC reagent 37
2.2.10 Blocking solution for immunohistochemistry 37
2.2.11 Solution for diaminobenzidine reaction 37
2.2.12 Thionine solution for Nissl staining 37
2.2.13 Hoechst olution 38
2.2.14 Blocking solution for double immunofluorescence 38
labeling

2.3 Animals 38
2.4 ntibodies 9
2.5 Immunohistochemistry 41
2.6 Double-labeling immunostaining 42
2.7 Nisl taing 42
2.8 Whole mount immunostaining 43
2.9 In situ hybridization 44
2.10 Flatmount double RNA in situ hybridization 45
6
2.11 Photomicrograph production 46

3 Results 47

3.1 Cadherin-7 expression in the spinal cord 47
3.2 Cadherin-7 expression in the hindbrain 51
3.3 Expression of cadherin-7 and cadherin-6B by 57
branchiomotor neurons
3.4 Postmigratory localization of branchiomotor neurons 60

4 Discussion 53

4.1 Expression of cadherin-7 marks the most dorsal portion of 64
the basal plate
4.2 Brachiomotor neurons assume their final positions in the 67
alar plate
4.3 Change of cadherin expression during branchiomotor 69
neuron migration and differentiation

5 Sumary 72

6 References 73
7
List of Abbreviations

1-6 rhombomeres 1-6
A alar plate
α α-catenin
B basal plate
β β-catenin
b/a basal/alar plate boundary
BCIP 5-bromo-4-chloro-3-indolyl phosphatetoluidine salt
Bl lateral part of basal plate
Bm medial part of basal plate
bm branchiomotor
cad6B cadherin-6B
cad7 cadherin-7
CNS central nervous system
d dorsal band of Phox2b expression
D/V dorsoventral
E embryonic day (day of incubation)
F floor plate
flm medial longitudinal fascicle
fp/FP floor plate
gV trigeminal ganglion
gVII facial ganglion
gIX glossopharyngeal ganglion
8
gX vagus ganglion
Hbhindbrain
HBSS HEPES-buffered salt solution
HH stage according to Hamburger and Hamilton (1951)
III oculomotor complex
IV trochlear complex
Isl1 Islet-1
mbm migrating branchiomotor neurons
MN motor neuron column
mz mantle zone
nc/NC notochord
NBT nitroblue tetrazolium salt
p0-p3 ventral progenitor cell domain
PBS phophate-buffered saline
pMN motorneuron progenitor domain
Rcad R-cadherin
RP roof plate
SG spinal ganglion
Shh Sonic hedgehog
s.m. somatomotor
s.s. somatosensory
TBS Tris-buffered saline
Tect tectum
Thio thionine
9
v4 fourth ventricle
Vm laterally/dorsally migrated branchiomotor nucleus of the
trigeminal nerve
Vmn ventral motor nucleus of the trigeminal nerve
v.m. visceromotor
VIn abducens nerve
VIIn facial nerve
VIIm laterally/dorsally migrated branchiomotor nucleus of the
facial nerve
vr ventral root
v.s. viscerosensory
vz ventricular zone
XII hypoglossal nucleus
XIIn hypoglossal nerve
Xm laterally/dorsally migrated branchiomotor nucleus of the
vagus nerve
10