Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex:
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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2008
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| Beschreibung: | 95 S. Ill., graph. Darst. 21 cm |
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Datensatz im Suchindex
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| adam_text | Titel: Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spin
Autor: Stavridis, Stavros I.
Jahr: 2008
TABLE OF CONTENTS
TABLE OF CONTENTS v
1. INTRODUCTION. 1
1.1 Spinal cord injury (SCI) 1
1.1.1. Epidemiology and demographics of SCI 1
1.1.2. PathophysiologyofSCI 2
1.1.2.1. Primary lesion 2
1.1.2.2. Secondary lesion 2
1.1.2.2.1. Vascular mechanisms 3
1.1.2.2.2. Free radicals 3
1.1.2.2.3. Apoptosis 3
1.1.3. Therapeutic interventions in SCI 4
1.2 CNS regenerative capacity 5
1.3 Neurotrophins........ 6
1.3.1 Neurotrophic and neuroprotective effects of NT-3 7
1.4. Experimental models 8
1.4.1. Organotypic slice cultures (OSCs) 8
1.4.2. The rat corticospinal tract (CST) 9
1.4.3. Axonal labeling 10
1.5. Immunohistochemical markers 10
1.5.1. NeuN 10
1.5.2. ChAT 11
1.5.3. Calbindin 11
1.5.4. GFAP 11
1.5.5. IB4 12
1.5.6. Nestin 12
1.6. Hypotheses 13
2. MATERIALS AND METHODS... 15
v
2.1. Materials 15
2.1.1. Substances, solutions and antibodies 15
2.1.2. Storage of substances and antibodies 16
2.1.3. Buffer solutions 16
2.1.4. Preparation medium and culture medium 17
2.1.5. Establishment of Agar-blocks 17
2.1.6. Devices 17
22. Methods 18
2.2.1. Preparation of rat spinal cord OSCs 18
2.2.2. Culture of OSCs 19
2.2.3. Fixation of OSCs 19
2.2.4. Immunohistochemistry 20
2.2.5. Statistical analysis 21
2.2.6. Preparation and culture of rat cortex and spinal cord
co-cultures 22
2.2.7. Subgroup division 23
2.2.8. Fixation of rat cortex and spinal cord cocultures 23
2.2.9. Immunohistochemistry 24
3. RESULTS 25
3.1. Part I: Characterization of spinal cord slice cultures (SCSCs) 25
3.1.1. Distribution of neuronal and glial cell populations in the
spinal cord (description of acutely fixed slices) 25
3.1.1.1. NeuN and ChAT- immunoreactivity 25
3.1.1.2. Calbindin-immunoreactivity 26
3.1.1.3. GFAP-immunoreactivity 26
3.1.1.4. IB4-immunoreactivity 26
3.1.1.5. Nestin-immunoreactivity 27
3.1.2. The spatiotemporal reaction pattern of neuronal and
glial cells in SCSCs during culture 27
3.1.2.1. Early motoneuron death 27
3.1.2.2. Calbindin* neurons persist in SCSCs 28
3.1.2.3. GFAP-immunoreactivity in SCSCs increases
early during the culture period 28
vi
3.1.2.4. ffi4-iminunoreactivity increases in the white matter.,29
3.1.2.5. Nestin-immunoreactivity persists in vitro and is
found in cells lining the central canal 29
3.2 Part II: Establishing a slice co-culture model of motorcortex and
spinal cord 44
3.2.1. Almost no sprouting axons are found in the control group 44
3.2.2. Effect of exogenous NT-3 application: Sprouting
axons are observed in the NT-3 group 45
4. DISCUSSION 56
Part I: Characterization of spinal cord slice cultures 56
4.1. Advantages and disadvantages of OSCs . 56
4.2. The fate of neuronal populations in SCSCs 57
4.2.1. Preservation of calbindin* neurons 57
4.2.1.1. The neuroprotective effect of calbindin 58
4.2.2. Early loss of large motoneurons in the ventral horn 58
4.2.2.1. The role of afferent connections in neuronal
survival 58
4.2.2.2. The role of targets in neuronal survival 59
4.2.2.3. The role of neurotrophins in motoneuronal survival
and developmental cell death 60
4.2.2.4. Necrosis and apoptosis 60
4.3. Reaction patterns of glial populations . ...— 61
4.3.1. Astrocytic reaction and glial scar formation after injury 61
4.3.1.1. Two-phase increase in GFAP-immunoreactivity 62
4.3.1.2. Reactive astrogliosis 62
4.3.1.3. The inhibitory effect of reactive astrogliosis
on axonal regeneration 63
4.3.1.4. The growth-promoting properties of astrocytes 63
4.3.2. The role of microglial cells 64
4.3.2.1. Microglial subpopulations 64
4.3.2.2. The concept of microglial functional plasticity 65
4.3.2.3. Microglial activation at the site of neuronal or
axonal damage 65
4.3.2.4. The spatiotemporal course of microglial
activation after injury 65
vii
4.4. Nestin and stem cells in the injured spinal cord. .66
4.4.1. Nestin expression in the spinal cord 66
4.4.2. Nestin upregulation and the role of stem cells after SCI 67
Part II: Establishing a slice co-culture model of motorcortex and spinal
cord.........68
4.5. Death of corticospinal neurons after lesion. The role of
age-and target dependency.. ..... ...69
4.6. The role of exogenous NT-3 application 69
4.6.1. The neuroprotective effect of NT-3 69
4.6.2. NT-3 promotes axonal regeneration and sprouting 70
4.7. Myelin-associated growth-inhibitors 70
4.8. Overview............72
5. SUMMARY .74
6. ZUSAMMENFASSUNG .75
7. LIST OF ABBREVIATIONS 77
8. BIBLIOGRAPHY .79
9. APPENDIX .89
9.1. Acknowledgement .89
9.2. Curriculum vitae 90
Q1 F ehpnslaiif 92
9.4. Honorary statement..........94
9.5 Ehrenwortliche Erklarung 95
viii
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| any_adam_object | 1 |
| author | Stavridis, Stavros I. 1974- |
| author_GND | (DE-588)137034385 |
| author_facet | Stavridis, Stavros I. 1974- |
| author_role | aut |
| author_sort | Stavridis, Stavros I. 1974- |
| author_variant | s i s si sis |
| building | Verbundindex |
| bvnumber | BV035318449 |
| ctrlnum | (OCoLC)314789063 (DE-599)DNB992273595 |
| dewey-full | 617.48 |
| dewey-hundreds | 600 - Technology (Applied sciences) |
| dewey-ones | 617 - Surgery & related medical specialties |
| dewey-raw | 617.48 |
| dewey-search | 617.48 |
| dewey-sort | 3617.48 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
| format | Thesis Book |
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| spelling | Stavridis, Stavros I. 1974- Verfasser (DE-588)137034385 aut Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex vorgelegt von Stavros I. Stavridis 2008 95 S. Ill., graph. Darst. 21 cm txt rdacontent n rdamedia nc rdacarrier Frankfurt (Main), Univ., Diss., 2008 (DE-588)4113937-9 Hochschulschrift gnd-content HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017123066&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Stavridis, Stavros I. 1974- Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex |
| subject_GND | (DE-588)4113937-9 |
| title | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex |
| title_auth | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex |
| title_exact_search | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex |
| title_full | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex vorgelegt von Stavros I. Stavridis |
| title_fullStr | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex vorgelegt von Stavros I. Stavridis |
| title_full_unstemmed | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex vorgelegt von Stavros I. Stavridis |
| title_short | Characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex |
| title_sort | characterization of postnatal rat spinal cord slice and studies on co cultures of postnatal rat spinal cord and motorcortex |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017123066&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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