Generation and characterisation of tau-transgenic mice:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2004
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | IX, 147 Bl. Ill., graph. Darst. 30 cm |
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| 100 | 1 | |a Zabke, Claudia |d 1971- |e Verfasser |0 (DE-588)129621374 |4 aut | |
| 245 | 1 | 0 | |a Generation and characterisation of tau-transgenic mice |c von Claudia Zabke |
| 264 | 1 | |c 2004 | |
| 300 | |a IX, 147 Bl. |b Ill., graph. Darst. |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
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| 502 | |a Berlin, Humboldt-Univ., Diss., 2004 | ||
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CONTENTS
CONTENTS
TABLE
OF
CONTENTS
.
ACKNOWLEDGEMENTS
.
V
ABBREVIATIONS
.
VI
ABSTRACT
.
VIII
ZUSAMMENFASSUNG
.
IX
1.
INTRODUCTION
.
1
1.1.
NORMAL
FUNCTIONS
OF
THE
TAU
PROTEINS
.
1
1.1.1.
GENE
ORGANISATION
AND
SPLICING
.
1
1.1.2.
STRUCTURE
AND
ROLES
OF
THE
TAU
PROTEIN
.
2
1.1.3.
POSTTRANSLATIONAL
MODIFICATIONS
.
4
1.2.TAUOPATHIES
.5
1.2.1
.ALZHEIMER
'
S
DISEASE
(AD)
.
5
1.2.2.
FAMILIAL
AD
(FAD)
.
10
1.2.3.
FRONTOTEMPORAL
DEMENTIA
WITH
PARKINSONISM
LINKED
TO
CHROMOSME
17
(FTDP-17)10
1.2.4.
OTHER
TAUOPATHIES
.
12
1.3.
MODELLING
TAU
AGGREGATION
.
12
1.4.
ANIMAL
MODELS
OF
AD
AND
FDTP-1
7
.
13
1.5.
AIM
OF
THE
STUDY
.
15
2.
MATERIALS
.
17
2.1.
CHEMICALS
.
17
2.2.
VECTORS
.
17
2.3.
DNA
SIZE
STANDARDS
.
17
2.4.
ENZYMES
AND
ANTIBODIES
.
17
2.5.
BACTERIAL
STRAINS
.
18
2.6.
ANIMALS
.
18
2.7.
KITS
.
18
2.8.
BUFFERS
.
19
2.9.
SOLUTIONS
FOR
BACTERIAL
CULTURE
.
21
2.10.
LIST
OF
PRIMERS
.
21
3.
METHODS
.
22
3.1.
GENERAL
CLONING
TECHNIQUES
.
22
3.2.
PREPARATION
OF
COMPETENT
E.
COLI
CELLS
FOR
TRANSFORMATION
.
22
3.2.1.
PREPARATION
OF
COMPETENT
E.
COLI
CELLS
FOR
HEAT
SHOCK
TRANSFORMATION
.
22
3.2.2.
PREPARATION
OF
CHEMICALLY
COMPETENT
E.
COLI
CELLS
FOR
TRANSFORMATION
.
23
CONTENTS
3.3.
TRANSFORMATION
OF
COMPETENT
E.
COLI
CELLS
.
23
3.3.1.
HEAT
SHOCK
METHOD
.
23
3.3.2.
TRANSFORMATION
OF
CHEMICALLY
COMPETENT
CELLS
.
23
3.4.
SMALL
SCALE
PREPARATION
OF
PLASMID
DNA
-
"
PLASMID
MINI-PREPS
"
.
24
3.4.1.
PROTOCOL
1
(SAMBROCK
ET
AL.,
1989)
.
24
3.4.2.
USE
OF
NUCLEOSPIN
PLUS
PLASMID
KIT
(BD
BIOSCIENCES
CLONTECH)
.
24
3.5.
LARGE
SCALE
PREPARATIONS
OF
PLASMID
DNA
-
"
PLASMID
MAXI-PREPS
"
.
24
3.6.
GENERATION
OF
TRANSGENIC
MICE
.
24
3.7.
GENOTYPING
OF
MICE
.
25
3.8.
PREPARATION
OF
GENOMIC
DNA
.
25
3.8.1.
PREPARATION
OF
GENOMIC
DNA
FOR
SOUTHERN
BLOT
ANALYSIS
.
25
3.8.2.
USE
OF
QIAGEN
TISSUE
KIT
.
26
3.9.
SOUTHERN
BLOT
.
26
3.10.
TOTAL
RNA
EXTRACTION
FROM
MAMMALIAN
TISSUE
.
27
3.10.1.
TRIZOL-METHOD
.
27
3.10.2.
ABSOLUTELY
RNAYY
RT-PCR
MINIPREP
KIT
(STRATAGENE)
.
27
3.11.
DNASE
DIGESTION
.
27
3.12.
NORTHERN
BLOT
.
28
3.13.
RADIOLABELLING
OF
DNA
.
28
3.14.
HYBRIDISATION
CONDITIONS
.
28
3.15.
CDNA
SYNTHESIS
.
29
3.16.
POLYMERASE
CHAIN
REACTION
(PCR)
.
29
3.17.
REVERSE
TRANSCRIPTION
PCR
ANALYSIS
.
30
3.18.
BLOTTING
OF
PCR
PRODUCTS
(ALKALI
BLOT)
.
30
3.19.
CLONING
OF
PCR
PRODUCTS
.
31
3.20.
LIGHTCYCLER
ANALYSIS
.
31
3.21.
RIBOPROBE
SYNTHESIS
.
32
3.22.
IN
SITU
HYBRIDISATION
.
33
3.23.
BRAIN
DISSECTION
.
34
3.24.
PARAFFIN
EMBEDDING
OF
MOUSE
BRAINS
.
35
3.25.
HAEMATOXYLIN
STAINING
.
35
3.26.
CRESYL
VIOLET
STAINING
.
36
3.27.
IMMUNOHISTOCHEMISTRY
.
36
3.28.
THIOFLAVIN-S
STAINING
.
37
3.29.
BIELSCHOWSKY
SILVER
STAINING
.
37
3.30.
SHIRPA-PROTOCOL
.
38
4.
RESULTS
.
40
II
CONTENTS
4.1.
CLONING
OF
THE
CONSTRUCTS
FOR
MICROINJECTION
.
40
4.1.1.
GENERAL
CONSIDERATIONS
.
40
4.1.2.
CLONING
OF
THE
THY295-390
CONSTRUCT
.
40
4.1.3.
CLONING
OF
THE
THY1
186-441
CONSTRUCT
.
42
4.1.4.
CLONING
OF
THE
P301S/G335DTHY
CONSTRUCT
.
42
4.2.
GENERATION
AND
IDENTIFICATION
OF
TRANSGENIC
MICE
.
43
4.3.
GENOMIC
SOUTHERN
ANALYSIS
OF
THE
DIFFERENT
TRANSGENIC
MOUSE
LINES
.
46
4.3.1.
ANALYSIS
OF
TRANSGENE
INTEGRATION
AND
GENERATION
OF
TRANSGENIC
MOUSE
LINES
.
46
4.3.2.
DETERMINATION
OF
THE
COPY
NUMBER
OF
THE
DIFFERENT
TRANSGENIC
LINES
.
50
4.4.
ANALYSIS
OF
RNA
.
50
4.4.1.
DETECTION
OF
RNA
EXPRESSION
IN
TRANSGENIC
MICE
USING
NORTHERN
BLOT
.
50
4.4.2.
DETECTION
OF
RNA
EXPRESSION
IN
TRANSGENIC
MICE
USING
RT-PCR
.
54
4.4.3.
ANALYSIS
OF
THE
ORIGIN
OF
THE
SECOND
TRANSCRIPT
OCCURING
IN
THE
SHORT
CONSTRUCT
.
56
4.4.4.
QUANTITATIVE
ANALYSIS
OF
RNA
.
58
4.4.5.
NON
RADIOACTIVE
IN
SITU
HYBRIDISATION
ANALYSIS
OF
LINE
1
AND
LINE
86B
.
62
4.5.
HISTOLOGICAL
ANALYSIS
.
70
4.5.1.
GENERAL
COMMENTS
.
70
4.5.2.
IMMUNOHISTOCHEMISTRY
.
70
4.5.3.
BIELSCHOWSKY
SILVER
STAIN
.
81
4.5.4.
THIOFLAVIN-S
STAINING
.
82
4.6.
BEHAVIOURAL
ANALYSIS
.
87
4.7.
PHENOTYPE
OF
P301S/G335D
MICE
.
91
5.
DISCUSSION
.
92
5.1.
GENERATION
OF
TRANSGENIC
MICE
.
92
5.1.1.
EXISTING
TRANSGENIC
MICE
RELEVANT
TO
ALZHEIMER
'
S
DISEASE
(AD)
.
92
5.1.2.
CONSTRUCTS
USED
FOR
MICROINJECTION
.
95
5.1.3.
GENERATION
OF
TRANSGENIC
MOUSE
LINES,
DNA
AND
RNA
ANALYSIS
.
98
5.2.
VARIEGATION
OF
EXPRESSION
SITE
AND
LEVEL
IN
ANIMALS
OF
LINE
1
.
99
5.2.1.
EXPRESSION
PATTERN
.
99
5.2.2.
EXPRESSION
LEVEL
.
100
5.3.
HISTOLOGICAL
AND
BEHAVIOURAL
ANALYSIS
.102
5.3.1.
LINE
1
AND
86B
.
102
5.3.2.
LINE
36
AND
66
.
107
5.4.
CONCLUSIONS
.
110
6.
REFERENCES
.
112
7.
APPENDIX
.
129
7.1.
SEQUENCES
OF
THE
CONSTRUCTS
USED
FOR
MICROINJECTION
.
129
III
CONTENTS
7.1.1.
SEQUENCE
295THY
.
129
7.1.2.
SEQUENCE
OF
1
86THY
.
132
7.1.3.
SEQUENCE
OF
THYP301S/G335D
.
135
7.2.
CELL
COUNTS
OF
TRANSGENIC
ANIMALS
.
138
7.3.
MUTATIONS
IN
THE
TAU
GENE
.
141
7.4.
CURRICULUM
VITAE
.
146
7.5.
ERKLSRUNG
.
147
IV |
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| author | Zabke, Claudia 1971- |
| author_GND | (DE-588)129621374 |
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| bvnumber | BV023123824 |
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| discipline_str_mv | Biologie Medizin |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV023123824 |
| illustrated | Illustrated |
| index_date | 2024-07-02T19:52:35Z |
| indexdate | 2024-07-09T21:11:36Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-016326261 |
| oclc_num | 634777442 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-11 |
| owner_facet | DE-355 DE-BY-UBR DE-11 |
| physical | IX, 147 Bl. Ill., graph. Darst. 30 cm |
| publishDate | 2004 |
| publishDateSearch | 2004 |
| publishDateSort | 2004 |
| record_format | marc |
| spelling | Zabke, Claudia 1971- Verfasser (DE-588)129621374 aut Generation and characterisation of tau-transgenic mice von Claudia Zabke 2004 IX, 147 Bl. Ill., graph. Darst. 30 cm txt rdacontent n rdamedia nc rdacarrier Berlin, Humboldt-Univ., Diss., 2004 Maus (DE-588)4169148-9 gnd rswk-swf cDNS (DE-588)4212297-1 gnd rswk-swf Tau-Protein (DE-588)4328997-6 gnd rswk-swf Transgene Tiere (DE-588)4279021-9 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Maus (DE-588)4169148-9 s Transgene Tiere (DE-588)4279021-9 s Tau-Protein (DE-588)4328997-6 s cDNS (DE-588)4212297-1 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016326261&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Zabke, Claudia 1971- Generation and characterisation of tau-transgenic mice Maus (DE-588)4169148-9 gnd cDNS (DE-588)4212297-1 gnd Tau-Protein (DE-588)4328997-6 gnd Transgene Tiere (DE-588)4279021-9 gnd |
| subject_GND | (DE-588)4169148-9 (DE-588)4212297-1 (DE-588)4328997-6 (DE-588)4279021-9 (DE-588)4113937-9 |
| title | Generation and characterisation of tau-transgenic mice |
| title_auth | Generation and characterisation of tau-transgenic mice |
| title_exact_search | Generation and characterisation of tau-transgenic mice |
| title_exact_search_txtP | Generation and characterisation of tau-transgenic mice |
| title_full | Generation and characterisation of tau-transgenic mice von Claudia Zabke |
| title_fullStr | Generation and characterisation of tau-transgenic mice von Claudia Zabke |
| title_full_unstemmed | Generation and characterisation of tau-transgenic mice von Claudia Zabke |
| title_short | Generation and characterisation of tau-transgenic mice |
| title_sort | generation and characterisation of tau transgenic mice |
| topic | Maus (DE-588)4169148-9 gnd cDNS (DE-588)4212297-1 gnd Tau-Protein (DE-588)4328997-6 gnd Transgene Tiere (DE-588)4279021-9 gnd |
| topic_facet | Maus cDNS Tau-Protein Transgene Tiere Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=016326261&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT zabkeclaudia generationandcharacterisationoftautransgenicmice |